Antioxidants doi: 10.3390/antiox15091164

Authors: Giuseppina Augimeri Mary Fucile Alberto Vaccarella Giancarlo Statti Filomena Conforti Daniela Bonofiglio

The Mediterranean Diet pattern is characterized by high intake of plant-based foods rich in bioactive nutrients, including carotenoids. This review examines the major dietary carotenoids and xanthophylls characteristic of the Mediterranean food matrix, focusing on their structural diversity, plant biosynthesis, release from the food matrix, bioavailability, and biological redox-modulating activities. The literature was collected from PubMed, Web of Science, and Scopus to trace the pathway of carotenoids from their accumulation in plant tissues to their potential effects on human health. Culinary processing and the concomitant intake of dietary lipids can enhance carotenoid bioaccessibility by promoting isomerization and facilitating the formation of mixed micelles during digestion. In addition to systemic absorption, noninvasive diagnostic tools, including skin carotenoid measurements by reflectance spectroscopy and macular pigment optical density, provide useful biomarkers of fruit and vegetable intake. Carotenoids may contribute to systemic antioxidant and redox regulation by quenching singlet oxygen, scavenging free radicals, and modulating redox-sensitive signaling pathways. Overall, their structural diversity, food-matrix interactions, bioaccessibility, and biological activities support continued investigation of carotenoid-rich foods within the Mediterranean dietary pattern and their associations with human health outcomes.

Antioxidants doi: 10.3390/antiox15091163

Authors: Akanksha R Gautam Soottawat Benjakul Avtar Singh Ravindran Muthukumarasamy Nilesh Nirmal Saleh Al-Ghamdi Mohammad Fikry

Shrimp shell–derived chitosan and protein hydrolysate–epigallocatechin gallate conjugates (SEC) were used to fabricate hybrid particles (C-SEC) at mass ratios of 1:1, 1:5, and 1:10 using an alkali precipitation method. SEC incorporation significantly enhanced the antioxidant capacity of chitosan particles (p < 0.05). Among the formulations, C-SEC prepared at a ratio of 1:5 (C–SEC–1:5) exhibited the highest antioxidant activities, including DPPH (14.55 mM TE/g) or ABTS (9.80 mM TE/g), or hydroxyl (70.29%) radical scavenging activities, ferric reducing antioxidant power (10.59 mM TE/g), and metal chelating activity (6.37 mM EE/g). The particle size was markedly reduced from 738 nm for chitosan particles (CP) to 112 nm following incorporation of SEC, indicating the formation of finely dispersed hybrid particles. 1H NMR analysis confirmed successful molecular integration of SEC into the chitosan matrix through non-covalent interactions. Thermal analyses revealed altered degradation and transition behavior of the hybrid particles. TGA showed that the principal degradation stages shifted to higher temperatures, whereas DSC showed an increase in transition enthalpy from 370.74 to 2449.60 J/g. SEM and TEM images revealed that the C–SEC–1:5 formed porous, interconnected particle networks with rough surfaces. The selected hybrid particles also showed superior inhibition of Cu2+–induced lecithin liposome oxidation and β-carotene bleaching compared with CP, demonstrating concentration-dependent protection against lipid oxidation. Overall, these findings indicate the potential of C–SEC–1:5 as an antioxidant-based functional ingredient and bioactive delivery vehicle in food and nutraceutical formulations.

Antioxidants doi: 10.3390/antiox15091162

Authors: Maria Luisa Valle Christopher Lawrence De Jesus Bitseat Getaneh Hediye Erdjument-Bromage James Loveland Christopher William Donatello Arienzo Thomas A. Neubert Agueda Rostagno Jorge Ghiso

Hyperhomocysteinemia (HHcy), a risk factor for cardiovascular, neurodegenerative, and metabolic dysfunctions in humans, has been recapitulated in murine models by genetic and dietary modifications. The relevance of HHcy in clinical and experimental settings has led to the development of various analytical techniques with varying sensitivity and sample volume requirements. This study uses a fit-for-purpose, small-volume LC-MS/MS workflow to quantify plasma homocysteine (Hcy), methionine (Met), and cysteine (Cys) in a diet-induced HHcy mouse model. Using these assays that overcome limitations imposed by sample availability in small animals, our study evaluated circulating levels of these amino acids in C57BL/6 mice fed a HHcy-inducing diet rich in Met and lacking vitamins B6, B9, and B12. The increased Hcy and decreased Cys plasma concentrations observed in these mice correlated with decreased glutathione concentrations in brain homogenates and concomitant increase in the activity of the antioxidant enzyme Glutathione-S-Transferase and in the levels of the lipid peroxidation marker malondialdehyde. Overall, our work links plasma HHcy to the induction of brain oxidative stress, supporting the role of dietary modifications as complementary therapeutic strategies for neurodegenerative conditions.

Antioxidants doi: 10.3390/antiox15091161

Authors: Mariarosaria Ingegneri Martina Imbesi Souda Belaid Marta Mangano Maria Neve Ombra Filomena Nazzaro Antonella Smeriglio Domenico Trombetta

Citrus processing by-products represent a sustainable source of bioactive plant complexes with potential applications in intestinal health. This study investigated the effects of standardized food-grade orange (OE) and lemon (LE) pomace extracts and their simulated gastrointestinal digestates (DIGs) on intestinal epithelial responses and selected probiotic strains. Caco-2 cells were used to assess cytotoxicity, epithelial wound closure, and, in differentiated monolayers challenged with lipopolysaccharide (LPS), extracellular levels of SOD2, catalase, Nrf2, IL-6, IL-8, TNF-α, and IL-1β. In parallel, the effects of OE, LE, and their corresponding DIGs on the growth and cell-surface hydrophobicity of four probiotic lactic acid bacteria were evaluated. OE and LE DIGs were non-cytotoxic and promoted epithelial wound closure in a concentration- and time-dependent manner. In LPS-challenged monolayers, both DIGs counteracted alterations in extracellular oxidative stress-related proteins and reduced pro-inflammatory cytokine levels without affecting cell viability. OE and LE also produced strain-dependent effects on probiotic growth and cell-surface hydrophobicity, which were modified by gastrointestinal digestion. Overall, these findings extend previous evidence on the intestinal bioactivity of Citrus pomace-derived plant complexes and support their further investigation as sustainable food-grade ingredients for gut health applications.

Antioxidants doi: 10.3390/antiox15091160

Authors: Shivani Bansal Sunain Deol Meth Jayatilake Yaoxiang Li Brian L. Fish Xiao Xu Jose A. Fernandez John H. Griffin Tracy Gasperetti Meetha Medhora Marjan Boerma Heather A. Himburg Amrita K. Cheema

Radiological emergencies necessitate biomarkers that not only estimate absorbed ionizing radiation (IR) dose but also guide timely interventions to prevent or delay multi-organ injury. Conventional LC–MS-based metabolomics of bulk plasma is constrained by matrix effects that mask low-abundance species. Extracellular vesicles (EVs) constitute a metabolically enriched, underexplored compartment that can provide complementary insight into systemic metabolic and redox responses to IR. Female WAG/RijCmcr rats were exposed to 13.0 Gy leg-out partial-body X-rays and treated with one of three activated protein C (APC) variants—rat wild-type (WT), rat 3K3A-APC, or human WT APC—administered 24- and 48 h post-irradiation. Longitudinal plasma collections (days 1, 14, 30, and 90) were subjected to metabolomic and lipidomic profiling of whole plasma and matched EV-enriched fractions to define signatures of acute radiation syndrome (ARS) and delayed effects of acute radiation exposure (DEARE), and their modulation by APC. ARS was marked by early dyslipidemia and widespread metabolic disruption, evolving into DEARE with persistent alterations in energy metabolism, and nucleotide biosynthesis, consistent with sustained oxidative and inflammatory stress. EV profiles showed matrix-specific, time-dependent trajectories distinct from plasma, with prominent lipid dysregulation and enrichment of fatty acid β-oxidation, sphingolipid, and cholesterol pathway metabolites at day 90. Rat 3K3A-APC promoted early EV metabolic normalization, whereas rat WT APC more effectively mitigated late DEARE-associated changes. Elevated sphingomyelins in plasma EVs at day 90 may suggest a compensatory or anti-inflammatory lipid response. These findings suggest that plasma-derived EVs may provide a sensitive matrix for radiation biomarker discovery and may help elucidate APC-mediated modulation of IR-induced metabolic and redox disturbances.

Antioxidants doi: 10.3390/antiox15091159

Authors: Natthrit Roekngam Wanninee Chankaew Suwichak Chaisit Sonsawan Kongpuckdee Sunisa Khongthong

Oxidative stress and chronic inflammation are major contributors to skin photoaging, highlighting the need for natural antioxidants with multifunctional bioactivities. This study developed an ultrasound-assisted extraction (UAE) strategy to recover an astaxanthin-rich extract from the underutilized freshwater macroalga Chara corallina and evaluated its antioxidant, anti-inflammatory, and anti-photoaging properties. Different solvent systems were evaluated for extraction efficiency, and astaxanthin-equivalent recovery was quantified by high-performance liquid chromatography with photodiode array detection (HPLC–PDA). Antioxidant activity was evaluated using the DPPH radical scavenging assay, whereas biological activities were assessed in human dermal fibroblasts and RAW264.7 macrophages using cell viability assays, quantitative real-time PCR, and extracellular matrix-related enzyme inhibition assays. The selected solvent system (48% ethanol in ethyl acetate) provided the highest astaxanthin-equivalent recovery (0.2598 ± 0.0086% w/w). The selected CCE exhibited DPPH radical-scavenging activity, modulated antioxidant- and inflammation-associated gene expression, increased COL1A2 mRNA expression, and inhibited collagenase, elastase, and hyaluronidase within the evaluated concentration range. HPLC–PDA analysis revealed a chromatographic component with retention-time and UV–visible spectral characteristics corresponding to those of an authentic astaxanthin reference standard; however, comprehensive structural and phytochemical characterization was not performed. Because CCE is a chemically complex extract, the observed biological responses cannot be attributed exclusively to astaxanthin. Overall, these findings provide an initial basis for further investigation of C. corallina as an underexplored freshwater source of carotenoid-containing bioactive extracts rather than establishing it as a commercially competitive source of natural astaxanthin.

Antioxidants doi: 10.3390/antiox15091158

Authors: Yilka Mena-Linares Humberto Vélez-Slimani Luis A. Salazar

Macrophage activation depends on the balance between pro-inflammatory signaling and cytoprotective antioxidant responses. Persistent nuclear factor kappa B (NF-κB) and inducible nitric oxide synthase (iNOS) activity, together with insufficient nuclear factor erythroid 2-related factor 2 (Nrf2) signaling, promotes oxidative and nitrosative stress and contributes to chronic inflammation. Phenylethanoid- and flavonoid-rich botanical extracts may modulate this immune-redox network, but the available evidence is dispersed across plant species, extraction procedures, isolated compounds, and experimental models. This review examines the phytochemical composition and biological effects of these preparations, with particular emphasis on Buddleja globosa Hope. Evidence concerning verbascoside, luteolin, apigenin, quercetin derivatives, and other constituents is integrated with findings related to toll-like receptor 4/NF-κB signaling, iNOS-derived nitric oxide, inflammatory cytokines, reactive oxygen species, and Keap1–Nrf2-dependent cytoprotective responses. Direct evidence obtained with B. globosa extracts is distinguished from mechanistic findings generated using isolated constituents or related botanical preparations. Current studies support antioxidant and inflammation-modulating activities, but interpretation is limited by phytochemical variability, inconsistent dosing, incomplete cytotoxicity assessment, and limited use of macrophage-specific models. Future work should prioritize chemically standardized extracts, direct extract–constituent comparisons, pathway-specific validation, and formulation strategies that improve stability, reproducibility, and biological performance.

Antioxidants doi: 10.3390/antiox15091157

Authors: Erkan Caner Ozkat

The Antioxidant Food Table is the largest open collection of measured food antioxidant capacity. It covers 3139 products assayed by the ferric reducing ability of plasma (FRAP) method. The table has served mainly as a dietary lookup source and has never been machine-readable or benchmarked. Here it was extracted into a validated open dataset (3135 records, 99.9%). A leakage-aware benchmark of antioxidant capacity prediction from product description and category was then constructed. Eighteen predictors, from naïve baselines to deep networks and fusions, were evaluated under two partitioning regimes with the same five seeds and permutation controls. Since 39.4% of records share a product name, the conventional random split rewards memorization. A learning-free duplicate lookup explained 45% of the apparent k-nearest-neighbor advantage over the category median. In grouped evaluation, ridge regression on term frequency–inverse document frequency (TF–IDF) features (R2=0.674) outperformed both deep networks. Pretrained word vectors did not close this gap. An equal-weight fusion of all eight models performed best (R2=0.684) with 2.6-fold lower variability. Protocol and representation, rather than architecture, dominated the outcome on this benchmark. The dataset, code, and predictions are released openly.

Antioxidants doi: 10.3390/antiox15091156

Authors: Giovanni Imbimbo Federica Foti Thomas Ammann Maria Grazia Chiappini Vittoria Cammisotto Valentina Castellani Pasquale Pignatelli Alessio Molfino

Background: Endotoxemia and oxidative stress may contribute to adverse body-composition changes in patients receiving maintenance hemodialysis, but their relationships with adiposity and muscularity remain incompletely defined. We investigated whether the concentrations of circulating lipopolysaccharide (LPS), soluble NOX2-derived peptide (sNox2-dp), and hydrogen peroxide (H2O2) showed differential associations with bioimpedance-derived body-composition compartments and explored their relationships with normalized protein catabolic rate (nPCR), a surrogate of protein intake. Methods: In this observational, cross-sectional study, adult patients receiving maintenance hemodialysis were enrolled at a single center. Serum LPS and sNox2-dp and H2O2 concentrations were measured before dialysis. Body composition was assessed by bioimpedance analysis; fat mass (FM) was used as an index of adiposity, whereas intracellular water indexed to height squared (ICW/h2) was used as a proxy of muscularity. Associations were examined using Spearman correlation and multivariable linear regression adjusted for age, sex, and nPCR. Results: A total of 58 participants were included with a median age of 73 years; 64% were male and mean body mass index was 24.6 ± 4.2 kg/m2. LPS correlated with sNox2-dp (rho = 0.420, p = 0.001), whereas sNox2-dp correlated with H2O2 (rho = 0.352, p = 0.007); the LPS–H2O2 association was borderline (rho = 0.259, p = 0.050). sNox2-dp correlated positively with fat-free mass, total body water, ICW, and ICW/h2, whereas LPS correlated with FM (rho = 0.305, p = 0.023). Participants with nPCR > 0.89 g/kg/day had higher LPS concentrations than those with lower nPCR (p = 0.015), and nPCR correlated with ICW/h2 (rho = 0.36, p = 0.007). In multivariable analysis, LPS remained independently associated with FM (β = 0.210, p = 0.046). nPCR was positively associated with ICW/h2 at the threshold of statistical significance (β = 1.393, p = 0.050), whereas sNox2-dp showed a nonsignificant positive trend (p = 0.060). Conclusions: In maintenance hemodialysis, endotoxemia and NOX2 activation showed differential associations with body composition: LPS with adiposity and sNox2-dp with muscularity. Higher nPCR was associated with both greater muscularity and higher LPS concentrations, suggesting a complex relationship between protein intake, the gut–oxidative-stress axis, and body composition.

Antioxidants doi: 10.3390/antiox15091155

Authors: Shao-Wen Weng Yu-Han Lin Shih-Hsuan Chen Tsu-Kung Lin Pei-Wen Wang Chia-Wei Liou

Mitochondrial DNA (mtDNA) haplogroups influence mitochondrial function and reactive oxygen species (ROS) production, potentially modulating susceptibility to metabolic disorders. This study investigated the associations between mtDNA haplogroups, systemic oxidative stress, and metabolic syndrome in 2486 Taiwanese individuals. Serum thiobarbituric acid-reactive substances (TBARS) and free thiols were measured as markers of oxidative and antioxidative status, respectively, while mtDNA haplogroups were determined using multiplex PCR and Luminex genotyping. Macrohaplogroups N (51%) and M (49%) were predominant, with haplogroups B, F, D, and M7 most frequently observed. Although the overall haplogroup distribution is genetically similar to populations in Southern China, the distribution among the Taiwanese population shows distinct differences. Haplogroup B was significantly associated with higher TBARS levels, lower thiol concentrations, and an increased prevalence of diabetes and metabolic syndrome (OR 1.34, p = 0.004). Individuals with metabolic syndrome exhibited higher oxidative stress and lower antioxidative biomarker levels than those without the syndrome, while a progressive imbalance in oxidative–antioxidative status was observed with increasing numbers of metabolic syndrome components. These findings suggest that mtDNA haplogroups, particularly haplogroup B, may contribute to metabolic syndrome susceptibility through modulation of systemic oxidative stress.

Antioxidants doi: 10.3390/antiox15091154

Authors: Cheng-Wu Yang Wen-Hua Chen Tzong-Shyuan Lee

Oxidative stress is a central and well-established driver of cardiovascular disease, contributing to mitochondrial dysfunction, endothelial injury, inflammatory activation, and progressive myocardial and vascular remodeling. Although the major endogenous sources of cardiovascular reactive oxygen species (ROS), including NADPH oxidases, mitochondrial electron transport chain leakage, and uncoupled nitric oxide synthase, are well characterized, an additional and underappreciated contributor has recently emerged: cytochrome P450 2D6 (CYP2D6), an enzyme classically regarded as a hepatic drug-metabolizing protein. Accumulating evidence indicates that CYP2D6 is expressed extrahepatically in cardiac, vascular, and neural tissue, where uncoupled catalytic cycling is proposed to generate ROS independently of its canonical xenobiotic-metabolizing role, although direct experimental evidence for this pathway in human cardiac and vascular tissue remains limited. CYP2D6-derived oxidative processes may interact with mitochondrial respiratory function, endothelial nitric oxide bioavailability, and redox-sensitive inflammatory pathways, potentially contributing to cardiovascular vulnerability under specific genetic or pathological conditions. Critically, the magnitude of this oxidative contribution is not fixed: it is dynamically shaped by inherited CYP2D6 genetic variation, with poor and ultra-rapid metabolizer phenotypes exhibiting divergent oxidative burden and pharmacokinetic vulnerability, and is further amplified by polypharmacy, multimorbidity, and inflammation-driven phenoconversion, whereby clinically expressed CYP2D6 activity diverges from inherited genotype in ways that intensify redox imbalance. These dynamics are particularly relevant in East Asian populations, where the decreased-function CYP2D6*10 allele is highly prevalent. In this narrative, hypothesis-generating review, we integrate evidence from pharmacogenomics, redox biology, and cardiovascular pharmacology to propose a conceptual framework that reframes CYP2D6 as a genetically and pharmacologically tunable node within cardiovascular redox biology. We further examine emerging redox biomarkers, multi-omics platforms, and AI-assisted modeling as translational strategies for capturing this dynamic oxidative risk in real time. This framework supports a shift from static genotype-guided prescribing toward oxidative-risk-informed, adaptive cardiovascular precision medicine. Importantly, our focus on CYP2D6 should not be interpreted as evidence that it is a major cardiovascular CYP isozyme or an established driver of cardiovascular pathology. Rather, CYP2D6 is examined here as a deliberately hypothesis-generating candidate whose unusually strong pharmacogenetic variability, clinically important cardiovascular drug substrates, dynamic susceptibility to phenoconversion, extrahepatic expression, and mechanistically plausible links to endogenous substrate metabolism and CYP-associated ROS generation provide a convergent rationale for focused investigation. The mechanistic framework proposed in this review has not yet been experimentally and prospectively validated and should not be applied directly to clinical decision-making without supporting clinical data.

Antioxidants doi: 10.3390/antiox15091153

Authors: Ylenia Spissu Maria Lauda Tomasi Carla Cossu Andrea Floris Emanuela Azara Gavina Rita Serra Irene Marchesi Francesco Paolo Fiorentino Gaia Rocchitta Riccarda Zappino Antonio Barberis

Grapevine (Vitis vinifera L.) leaves are an abundant agricultural by-product and an underexploited source of bioactive polyphenols. This study characterized hydroalcoholic leaf extracts from autochthonous Sardinian cultivars and evaluated their effects in RKO human colorectal carcinoma cells. LC–HRMS revealed flavonol-rich profiles dominated by glycosylated derivatives of quercetin, kaempferol, and isorhamnetin, with marked cultivar-dependent differences and higher total flavonol abundance in Cannonau and Nieddera. Cyclic voltammetry showed distinct redox reactivity among extracts, with Vermentino and Martellada Bianca displaying the strongest electrochemical responses, consistent with their modulation of intracellular reactive oxygen species. In RKO cells, the extracts reduced metabolic activity in a dose-dependent manner, with IC50 values ranging from 100.3 to 251.8 µg/mL. At the molecular level, treatments modulated c-myb expression and shifted the bax/bcl2 balance toward a pro-apoptotic profile. These effects were supported by DAPI staining, showing chromatin condensation and nuclear fragmentation, and by moderate activation of caspase-3/7. Overall, Sardinian grapevine leaf extracts showed cultivar-specific phenolic composition, redox activity, and pro-apoptotic effects in an in vitro colorectal cancer model, supporting their potential valorization as sustainable sources of bioactive compounds. Further studies addressing bioavailability, metabolism, and in vivo effects are required to assess their physiological relevance and translational potential.

Antioxidants doi: 10.3390/antiox15091152

Authors: Aleyma Veliz Perez Mihaela Badea Sehrish Bilal

Cardiovascular disease (CVD) remains the leading cause of mortality worldwide, with aging as one of its main risk factors. Cellular senescence and oxidative stress form a bidirectional vicious cycle that drives vascular aging, endothelial dysfunction, and atherosclerotic progression. This state-of-the-art review synthesizes current evidence on the interplay between oxidative stress and senescence in cardiovascular aging and critically assesses whether antioxidant strategies can benefit vascular health. A targeted literature search was conducted in PubMed/MEDLINE, Web of Science, and Scopus (2016–2026). While endogenous antioxidant defenses decline with age, exogenous antioxidants, including resveratrol, vitamins C and E, omega-3 fatty acids, carotenoids, and coenzyme Q10,demonstrate promising preclinical effects, yet large-scale clinical trials have yielded inconsistent results. The VITAL and STRENGTH trials failed to demonstrate significant cardiovascular benefit with omega-3 supplementation, and elevated serum β-carotene was found paradoxically associated with increased cardiovascular mortality. Emerging mitochondria-targeted antioxidants (MitoQ, MitoTEMPO) show preclinical promise but require further clinical validation. Current evidence does not support antioxidant supplementation for cardiovascular prevention. Lifestyle interventions, particularly antioxidant-rich dietary patterns such as the Mediterranean diet, remain the safest strategy. Future research should develop personalized approaches guided by oxidative stress biomarkers and long-term trials.

Antioxidants doi: 10.3390/antiox15091151

Authors: Alberto Vázquez-Jiménez Margarita M. Marques José M. Sánchez Jesús Agulla Rebeca Lapresa Mónica Trigal-Martínez Rosalía Fernández-Alonso Gracia Merino Antonio Fernández Antonella Consiglio Juan P. Bolaños Ángeles Almeida María C. Marín Lorena López-Ferreras

Parkinson’s disease (PD) is characterized by oxidative stress, mitochondrial dysfunction, and dopaminergic neuron loss, for which effective treatments remain unavailable. Here, we report CL0179, a fungal-derived decahydrofluorene alkaloid with antioxidant-associated neuroprotective properties, and evaluate its effects across cellular and animal PD models. CL0179 exhibited a favorable safety profile and protected SHSY5Y against 6-hydroxydopamine- (6-OHDA), rotenone-, and 1-Methyl-4-phenylpyridinium-iodide (MPP+)-induced neurotoxicity by preserving mitochondrial membrane potential and network integrity. Transcriptomic analyses revealed selective restoration of gene-expression programs associated with oxidative phosphorylation, mitochondrial bioenergetics, and stress adaptation disrupted by MPP+. CL0179 also enhanced SIRT1 activity under MPP+ stress, whereas pharmacological SIRT1 inhibition partially attenuated protection of mitochondrial membrane potential and cell viability. In LRRK2-G2019S astrocytes, CL0179 reduced ROS and α-synuclein accumulation and restored mitochondrial organization, while in human dopaminergic neurons, it attenuated toxin-induced mitochondrial depolarization and preserved neuronal architecture. To overcome the low production of CL0179, we generated the structurally related analogue CL0670. Both compounds crossed the blood–brain barrier and protected mouse primary cortical neurons, while CL0670 improved motor deficits in a 6-OHDA mouse model. Collectively, these compounds promote mitochondrial resilience and stress-adaptive neuroprotection, supporting their potential for PD and related neurodegenerative disorders.

Antioxidants doi: 10.3390/antiox15091150

Authors: Fabio Lauria Paola Russo Alfonso Siani Ivana Sirangelo Ilenia D’Orsi Pasquale Marena Antje Hebestreit Ronja Foraita Giuseppe Iacomino

The widespread consumption of ultra-processed foods increases exposure to advanced glycation end-products (AGEs), key mediators of oxidative stress and chronic low-grade inflammation. Although AGEs contribute to adult metabolic dysfunction, their early molecular correlates in pediatric populations remain insufficiently defined. Extracellular vesicle (EV)-associated miRNAs represent stable, non-invasive indicators of systemic stress responses. This study evaluated the association between urinary AGEs and urinary EV-associated miRNAs in 94 Italian children and adolescents from the I.Family cohort. Fluorescent AGEs were quantified via spectrofluorimetry, and EV-enriched urinary miRNAs were profiled using next-generation sequencing. Differential expression and multivariable generalised linear models (adjusted for age, sex, BMI z-score, and hs-CRP) were performed. Differential expression analysis revealed a significant upregulation of hsa-miR-4516 in participants with high versus low urinary AGE levels (fold change = 2.31; FDR = 0.024). In multivariable continuous models, hsa-miR-4516 remained the primary AGE-associated miRNA, though the association attenuated following adjustment for BMI z-score and hs-CRP. Functional enrichment and network analyses highlighted pathways governing oxidative stress responses, proteostasis, and cellular survival. Overall, urinary EV-associated miRNAs may reflect coordinated biological responses to dietary AGE burden rather than serving as direct exposure biomarkers, offering integrated, non-invasive insights into early metabolic and inflammatory adaptations in children.

Antioxidants doi: 10.3390/antiox15091149

Authors: Zhanar Nabiyeva Akerke Kulaipbekova Asylbek Kozybayev Handi Yang Fuhang Song Abdyssemat Samadun Elmira Assembayeva Nasi Ai

A systematic phytochemical investigation of the leaves of Crataegus almaatensis Pojark., an endemic hawthorn species native to the mountainous regions of Kazakhstan, was carried out. Multi-step chromatographic fractionation, comprising ultrasound-assisted methanol extraction, sequential liquid–liquid partitioning, silica gel column chromatography, and preparative HPLC, afforded five phenolic compounds as individual, spectroscopically homogeneous constituents from the ethyl acetate fraction. The structures of all isolated compounds were elucidated by comprehensive analysis of high-resolution mass spectrometry (HR-ESI-QTOF-MS) data and one- and two-dimensional NMR spectroscopy (1H, 13C, HSQC, HMBC) in DMSO-d6. The following compounds were identified: (−)-epicatechin, cinchonain Ia, a methoxy- and hydroxy-substituted diaryl ether β-D-glucopyranoside, kakispyrol, and a dimethoxy-substituted diaryl ether β-D-glucopyranoside. The isolated compounds belong to three chemical classes: flavanols, flavolignans, and diaryl ether-type phenolic glycosides. Evaluation of ABTS and DPPH radical-scavenging activity revealed compound-specific antioxidant profiles, with compound 5 (cinchonain Ia) showing activity comparable to the ascorbic acid standard. This study provides, for the first time, a detailed structural characterization of individual phenolic metabolites from C. almaatensis leaves and makes a significant contribution to understanding the chemical diversity of this endemic species.

Antioxidants doi: 10.3390/antiox15091148

Authors: Hanna Schlemminger Swantje Melina Lockowandt Alina Löser Anna Patricia Kipp Lars-Oliver Klotz Holger Steinbrenner

Human selenium-binding protein 1 (SELENBP1) is a methanethiol oxidase (MTO), converting methanethiol (MT) to hydrogen sulfide (H2S), hydrogen peroxide (H2O2) and formaldehyde (HCHO). SELENBP1 has orthologs in all domains of life. RdMTO, an orthologous enzyme recently identified in the marine bacterium Roseobacter denitrificans, was postulated to require the cysteine residue closest to its C-terminus, Cys448, for MT binding and to oxidize MT to sulfane sulfur (S0) rather than to H2S. SELENBP1 and RdMTO exhibit ~53% sequence identity, with Cys448 (numbered Cys466 in SELENBP1) and amino acids required for copper binding being conserved. Therefore, we here compared the MTO activity of recombinant SELENBP1 and RdMTO. We found that SELENBP1, like RdMTO, converts MT as well as structurally related alkyl thiols to form H2S, H2O2 and, in the case of MT, HCHO in a strictly copper-dependent manner. MTO activity of both proteins was lowered but not abrogated upon mutation of their respective C-terminal cysteine residue. Thus, the catalytic mechanism of selenium-binding proteins that act as copper-dependent MTOs appears to be evolutionarily conserved from bacteria to humans. Presumably, this enzyme is an early evolutionary invention of prokaryotes, in order to cope with toxic thiols in their environment.

Antioxidants doi: 10.3390/antiox15091147

Authors: Sophie Wallace Spencer Lawes Adrienne C. Scheck Richard E. Frye

Copper is a redox-active transition metal that is essential for the assembly and catalytic function of cytochrome c oxidase (Complex IV), the terminal enzyme of the mitochondrial electron transport chain and a principal site of physiological oxygen reduction. Elevated Complex IV activity and respiratory chain uncoupling are among the most consistently replicated biological findings in autism spectrum disorder (ASD), yet the interaction between mitochondrial copper delivery, respiration, and cellular oxidant handling in ASD has not been systematically defined. We treated dermal fibroblasts from 9 children with ASD and 10 typically developing controls with the copper(I)–niacin complex Cunermuspir (0, 50, or 100 µM for 1 or 24 h exposure) and challenged them with graded concentrations (0–5.0 µM) of the redox-cycling agent 2,3-dimethoxy-1,4-naphthoquinone (DMNQ). Mitochondrial respiration was profiled by Seahorse XF respirometry (2133 observations across 28 experiments), and cellular reactive oxygen species (CellROX™ Green) and mitochondrial mass/polarization (MitoTracker™ Deep Red) were quantified by fluorescence imaging. ASD fibroblasts displayed a hypermetabolic, uncoupled respiratory phenotype (~73% higher baseline respiration; ~109% higher proton leak; reduced coupling efficiency). Linear mixed models with polynomial dose terms revealed significant ASD × Cunermuspir complex interactions (ASD × Cunermuspir and/or their higher-order interactions with DMNQ and treatment) for four respiratory parameters. ASD cells exhibited lower steady-state oxidation-dependent CellROX™ Green fluorescence than controls despite greater respiratory uncoupling, as well as lower steady-state MitoTracker™ Deep Red fluorescence; Cunermuspir reshaped the MitoTracker™ DMNQ dose–response in an ASD-selective manner. These findings identify the Cunermuspir-associated modulation of the ASD mitochondrial and oxidant handling phenotype and motivate further mechanistic and translational evaluation.

Antioxidants doi: 10.3390/antiox15091146

Authors: Sandy Peltier Théo Michel Fanny Mialane Mickaël Marin Michaël Dussiot Céline Rodriguez Monika Dzieciatkowska Marie Tamagne Camille Roussel Stéphanie Vicca Olivier Hermine Pierre A. Buffet Benoit Vingert Steven L. Spitalnik Angelo D’Alessandro Michel Prudent Pascal Amireault

Pre-transfusion storage of red blood cells (RBCs) induces aging in vitro driven by metabolic and oxidative stress, limiting transfusion efficacy. Unlike nucleated cells where multiple hallmarks characterize aging, proteostasis is expected to play a main role in anucleate RBCs, including proteasomal protein degradation. Although proteasomal activity declines during aging in vitro, its role in generating downstream alterations and post-transfusion clearance remains unclear. We hypothesized that proteasome inhibition accelerates RBC aging in vitro, particularly following re-exposure to physiological conditions. We evaluated the impact of proteasome inhibition (i.e., epoxomicin) on RBC quality during storage and physiological restoration in vitro. Additionally, young and old RBC subpopulations were compared. Proteasome inhibition during storage depleted ATP and altered RBC morphology without immediate oxidative damage. Physiological restoration of proteasome-inhibited RBCs caused accelerated ATP depletion, massive protein aggregation, reduced deformability, hemolysis, and phosphatidylserine exposure, particularly in long-stored RBCs. Strikingly, RBCs aged in vivo also exhibited low proteasomal activity and behaved similarly to stored RBCs following physiological restoration. In conclusion, proteasomal dysfunction is a key hallmark of RBC aging and senescence, driving molecular and cellular modifications that mark RBCs for clearance in vivo. Therefore, enhancing proteasomal function could improve RBC storage quality and transfusion efficacy.

Antioxidants doi: 10.3390/antiox15091145

Authors: Jovana Mišković Milena Rašeta Gordana Gojgić-Cvijović Marko Radenković Nenad Krsmanović Nemanja Živanović Gordana Ćirić-Marjanović Maja Milojević-Rakić Gordana Tamindžić Maja Karaman

This study investigated the biostimulant potential of endo- and exo-polysaccharides (PSHs) obtained from two Schizophyllum commune strains originating from Italy (ITA) and Serbia (SRB), with a primary focus on their ability to modulate the antioxidant defense and biochemical responses of Pisum sativum L. seedlings under optimal and drought conditions. The PSH fractions were structurally characterized by complementary spectroscopic and microscopic approaches, revealing strain- and drying-dependent differences in their structural features. The characterized PSHs were subsequently applied as seed biopriming agents, and pea seedlings were grown under two conditions: optimal growth and drought stress. Their effects were evaluated by analyzing enzymatic and non-enzymatic antioxidant responses, together with selected physiological and biochemical traits. Among the tested treatments, exo-PSHs differentially modulated antioxidant enzymes, with SRB primarily enhancing peroxidase and glutathione peroxidase and ITA increasing ascorbate peroxidase and catalase activity. SRB PSHs, particularly exo-PSH, showed the most pronounced effects on antioxidant-related responses, enhancing ABTS scavenging activity and altering DPPH capacity by approximately 35% compared with the control. In addition, endo-PSH from the ITA strain increased chlorophyll content by 26.12% and proline levels by 1.5-fold, indicating a distinct strain-dependent effect on plant physiological responses, particularly under stress conditions. LC–MS/MS profiling of plant MeOH extracts revealed a diverse range of secondary metabolites, including phenolic compounds, e.g., liquiritigenin, ferulic, and protocatechuic acids, which may contribute to the observed antioxidant responses. Multivariate analysis further supported clear biochemical differentiation among PSH treatments and growth conditions, confirming that the responses were dependent on both PSH origin and environmental conditions. Overall, these findings demonstrate that structurally distinct S. commune PSHs can differentially modulate antioxidant defense and key biochemical traits in pea seedlings, supporting their potential as sustainable fungus-derived biostimulants for enhancing plant resilience under both optimal and drought conditions.

Antioxidants doi: 10.3390/antiox15091144

Authors: Cristina Adriana Rosan George Alin Opris Alexandra Cristina Tocai (Moțoc) Daniela Gitea Ruben Budău Manuel Alexandru Gitea Simona Ioana Vicas

Allium ursinum L. (wild garlic) is a promising source of bioactive compounds for functional foods, nutraceuticals, and clean-label products. This review integrates current evidence on its phytochemical composition, biological activities, and food applications. The species contains sulfur compounds, polyphenols, vitamins, minerals, and pigments, whose levels vary with plant organ, developmental stage, environmental conditions, and processing. These constituents are associated mainly with antioxidant, antimicrobial, cardioprotective, and cytoprotective effects; however, evidence is derived predominantly from in vitro and animal studies, with limited clinical validation. In food systems, A. ursinum may enhance nutritional value, oxidative stability, sensory properties, and shelf life. Extraction, encapsulation, and stabilization technologies may support its use as a natural alternative to synthetic additives. Nevertheless, compositional variability, processing stability, bioavailability, authenticity, quality control, and raw-material supply remain major barriers to standardization and industrial application. Further research should combine phytochemical characterization with technological, safety, clinical, and sustainability assessments to enable reliable use of A. ursinum in food and nutraceutical products.

Antioxidants doi: 10.3390/antiox15091143

Authors: Nuriye Nuray Ulusu

Background: Obesity is a multifaceted metabolic disorder characterized by systemic disturbances, particularly impaired energy homeostasis, chronic low-grade inflammation, and mitochondrial dysfunction across the brain, gut, adipose tissue, and liver axes. Objectives: This review aims to examine the metabolic properties and molecular mechanisms of six key phytochemicals (berberine, resveratrol, catechins, capsaicin, thymoquinone, and phycocyanin) and the exercise-induced myokine irisin, and their roles in mitochondrial signaling and metabolic reprogramming. Sources of Evidence: A comprehensive literature search was conducted across major electronic databases, including PubMed, Web of Science, and Scopus, to identify relevant mechanistic, in vivo, and in vitro studies. Results: Both the selected phytochemicals and irisin act as multi-target regulators that modulate key signaling pathways, including AMPK, PI3K/Akt/mTOR, SIRT1, Nrf2, and PPARγ. These phytochemicals and irisin can drive cell- and tissue-specific metabolic reprogramming, promoting the browning of white adipocytes, suppressing de novo lipogenesis in hepatocytes, and enhancing fatty acid oxidation in skeletal myocytes. This synergistic metabolic reprogramming enhances thermogenesis and increases energy expenditure. Conclusions: Co-targeting redox signaling and metabolic pathways via phytochemicals and irisin offers a powerful strategy against obesity. This integrative framework restores multi-organ homeostasis, laying the groundwork for targeted metabolic therapies.

Antioxidants doi: 10.3390/antiox15091142

Authors: Argyrios Ginoudis Dimitra Pardali Mathios E. Mylonakis Serafeim Chaintoutis Dimitris Galiatsatos Angelos-Lauris Thomas Zoe Polizopoulou

Canine mast cell tumors (MCTs) exhibit heterogeneous biological behavior and provide a clinically relevant setting in which relationships between tumor phenotype and systemic redox homeostasis can be investigated. This exploratory prospective cohort study evaluated complementary circulating redox-related biomarkers representing hydroperoxide-derived oxidant products, antioxidant capacity, lipid peroxidation-related products, and oxidative DNA damage, and examined their associations with clinicopathological features of canine MCTs. Sixty-seven dogs with cytologically and histologically confirmed cutaneous or subcutaneous MCTs were included. Clinical staging was performed using cytological evaluation of lymph nodes, liver and spleen, and excised tumors were classified according to the Patnaik and Kiupel systems. Proliferative activity was assessed using Ki-67, and surgically excised lymph nodes were classified according to the Weishaar HN system. Serum reactive oxygen metabolites (d-ROMs), malondialdehyde (MDA), 8-hydroxy-2′-deoxyguanosine (8-OHdG) and antioxidant capacity using the OXY-adsorbent test were measured before therapeutic intervention. In the unadjusted analysis, serum MDA concentrations were higher in dogs with Ki-67 >23 than in dogs with Ki-67 ≤23 (median 10.8 vs. 7.0 μmol/L; raw p = 0.0254); however, this comparison did not remain statistically significant after Bonferroni correction for the 12 formal inferential biomarker × clinicopathological comparisons (adjusted p = 0.3052). The estimated Hodges–Lehmann location shift was +3.00 μmol/L (unadjusted bootstrap 95% CI, 0.58–6.00). No evaluated biomarker–clinicopathological association remained statistically significant after multiplicity adjustment. The observed MDA–Ki-67 pattern should therefore be regarded as exploratory and hypothesis-generating rather than as evidence of established prognostic or clinical utility. Long-term data on recurrence, progression, and survival were not available; therefore, prognostic utility could not be evaluated. Larger prospective studies incorporating longitudinal outcomes are warranted.

Antioxidants doi: 10.3390/antiox15091141

Authors: Elena-Andreea Moaleș Lucia Corina Dima-Cozma Cristina Andreea Adam Doina-Clementina Cojocaru Cristina Gena Dascălu Andreea Iațentiuc Iustin Mihai Iațentiuc Tatiana Drâmbă Maura Gabriela Felea Robert Negru Ioana Mădălina Zota Romică Sebastian Cozma Maria Magdalena Leon Florin Mitu

Chronic obstructive pulmonary disease (COPD) is increasingly recognized as a systemic disease characterized by chronic inflammation and oxidative stress, with frequent cardiovascular involvement. This study investigated the associations between oxidative stress-related inflammatory markers and structural and electrical cardiac abnormalities in patients with COPD using a comprehensive multimodal cardiopulmonary assessment. A total of 100 clinically stable patients with COPD underwent inflammatory biomarker evaluation (IL-6, IL-8, IL-1β, and TNF-α), transthoracic echocardiography and 24 h Holter ECG monitoring. Advanced COPD was associated with significant differences in several structural cardiac parameters, including left atrial volume (p = 0.016), left atrial height (p = 0.027), and selected left ventricular parameters, while left ventricular ejection fraction remained preserved. Electrical abnormalities were significantly associated with systemic inflammatory markers. SII showed the highest discriminatory performance for sinus rhythm abnormalities (AUC = 0.736), whereas IL-6 showed the highest discriminatory performance for ventricular premature complexes (AUC = 0.732). Ventricular ectopic activity was also associated with significantly higher SII, SIRI, MLR, NLR, and PLR. Systemic inflammatory markers were associated with structural and electrical cardiac abnormalities in COPD, suggesting that they may provide complementary information regarding cardiovascular involvement. However, given the cross-sectional design and moderate discriminatory performance of these biomarkers, the findings should be considered exploratory and do not establish causal relationships. Prospective studies are needed to determine their potential role in cardiovascular risk stratification.

Antioxidants doi: 10.3390/antiox15091140

Authors: Jan Krekora Jarosław Drożdż Janusz Blasiak Kai Kaarniranta

Increasing epidemiological and experimental evidence suggests that age-related macular degeneration (AMD) and cardiovascular disease (CVD) share multiple pathogenic mechanisms. Among these, oxidative stress has emerged as one of the most plausible links connecting retinal degeneration and cardiovascular pathology. Excessive production of reactive oxygen and nitrogen species, combined with declining antioxidant defenses, contributes to lipid peroxidation, mitochondrial dysfunction, chronic inflammation, complement activation, cellular senescence, and impaired cellular stress responses in both the retina and the vascular system. Notably, drusen (AMD) and atherosclerotic plaques (CVD) share several molecular constituents. Therefore, AMD and CVD may represent tissue-specific manifestations of broader age-related disturbances in redox homeostasis and inflammatory regulation. Nevertheless, the coexistence of AMD and CVD is incomplete, suggesting that genetic susceptibility, tissue-specific responses to oxidative stress, biological aging, and mechanisms of cellular resilience influence disease expression. In this review, we summarize current evidence linking AMD and CVD, examine oxidative stress-driven molecular pathways common to both disorders, discuss emerging biomarkers and therapeutic targets, and highlight important unresolved questions regarding disease heterogeneity and causal relationships. A better understanding of the shared mechanisms underlying AMD and CVD may facilitate the development of integrated preventive strategies, improved risk stratification, and more personalized therapeutic approaches for age-related diseases.

Antioxidants doi: 10.3390/antiox15091139

Authors: Rui Jia

Sustainable aquaculture demands not only high production efficiency but also the maintenance of physiological health in farmed aquatic animals [...]

Antioxidants doi: 10.3390/antiox15091138

Authors: Md Areeful Haque Mohammad Atiqur Rahman Inavolu Sriram Sandeep Sindhura Pasham Sayantap Datta Krishna M. Boini Saisudha Koka

Trimethylamine-N-oxide (TMAO), a gut microbiota-derived metabolite, is a known risk factor for cardiovascular disease. We previously showed that TMAO induces NLRP3 inflammasome activation and contributes to endothelial dysfunction. However, the upstream mechanisms linking TMAO to inflammasome activation and barrier injury remain unclear. In the present study, we examined whether membrane raft (MR) redox signaling contributes to TMAO-induced NLRP3 inflammasome activation and endothelial dysfunction. MR clustering, colocalization of MRs with NADPH oxidase subunits, inflammasome formation, and junction protein expression were assessed by Western blot analysis. RT-qPCR was performed to further assess the mRNA expression of junctional genes. Superoxide production was measured by electron spin resonance (ESR), caspase-1 activity was determined using a biochemical assay kit, IL-1β production was measured by ELISA, and endothelial permeability was evaluated using an FITC-dextran assay. TMAO increased MR clustering in endothelial cells in a dose-dependent manner. TMAO also enhanced the colocalization of MRs with p47phox and gp91phox, indicating the formation of an MR-associated redox signaling axis. In addition, TMAO increased superoxide production; promoted NLRP3 inflammasome formation; elevated caspase-1 activity and IL-1β production; reduced the expression of ZO-2, ZO-1, VE cadherin and occludin; and increased endothelial permeability. Pretreatment with the MR disruptor MCD, the NADPH oxidase inhibitor DPI, or the caspase-1 inhibitor WEHD significantly attenuated these TMAO-induced effects. These findings demonstrate that TMAO-mediated endothelial injury is dependent on the MR-associated redox signaling axis. TMAO promotes MR-associated redox signaling, leading to NADPH oxidase activation, superoxide generation, NLRP3 inflammasome activation, disruption of endothelial junction proteins, and barrier dysfunction. Targeting MR-associated signaling pathways may offer a novel therapeutic strategy to mitigate TMAO-induced vascular inflammation and endothelial dysfunction.

Antioxidants doi: 10.3390/antiox15091137

Authors: Po-Jen Hsiao Ching-Tsai Hsu Wen-Fang Chiang Jenq-Shyong Chan Li-Yen Huang Chung-Chi Yang Kuo-Cheng Lu

Chronic kidney disease–mineral and bone disorder (CKD-MBD) confers a substantial fracture burden that is only partly addressed by therapies targeting phosphate, parathyroid hormone, and vitamin D. Redox dysregulation may represent a complementary mechanism: reactive oxygen species (ROS) are required for receptor activator of nuclear factor-κB ligand (RANKL)-dependent osteoclastogenesis, whereas excessive ROS impair Wnt/β-catenin signalling in osteoblast precursors and promote osteocyte dysfunction. Uremic toxins, inflammation, and dialysis further increase oxidative stress. Molecular hydrogen (H2) is a highly diffusible redox modulator that has been proposed to limit damaging radical-chain reactions while preserving physiological oxidant signalling. In non-uremic skeletal models, H2 consistently suppresses osteoclast differentiation and bone loss, but evidence for osteoblast rescue is heterogeneous. In CKD and dialysis, H2-based interventions have shown signals of reduced oxidative stress and symptomatic benefit; however, human evidence is predominantly observational, and no study identified in this review assessed a bone-specific endpoint. We therefore integrate uremic bone redox biology with H2 pharmacology and propose a turnover-state-dependent model in which H2 may restrain excessive resorption in high-turnover disease, while its net effect in low-turnover adynamic bone remains uncertain because potential osteoblast rescue competes with anti-osteoclastic activity established only in non-uremic models. H2 should therefore be considered an experimental, mechanistically differentiated strategy requiring direct evaluation in uremic models and turnover-stratified clinical trials with parallel skeletal and vascular safety endpoints.

Antioxidants doi: 10.3390/antiox15091136

Authors: Adam R. Markowski Karolina Stępniak Piotr Zabielski Hady Razak Hady Aleksander Łukaszewicz Paulina Głuszyńska Patrycja Sadowska Urszula Chlabicz Agnieszka Błachnio-Zabielska

Colorectal cancer (CRC) exhibits substantial metabolic heterogeneity, but the organization of sphingolipid remodeling remains incompletely understood. In this exploratory single-center study, we integrated patient-matched tissue lipidomics, systemic oxidative stress profiling, and independent transcriptomic analyses. Tumor tissue, adjacent non-neoplastic mucosa, and preoperative serum were collected from 40 patients with CRC, with serum obtained from 23 hospitalized non-cancer controls. Sphingolipids were quantified by UHPLC–MS/MS, while total antioxidant capacity, total oxidant status, and oxidative stress index characterized systemic redox status. Paired lipidomics revealed coordinated remodeling with increased S1P-associated measures, selective ceramide depletion, and elevated S1P-to-ceramide ratios. Multivariate analyses did not identify stable discrete lipidomic subgroups and revealed variation consistent with a continuum-like organization within the measured sphingolipid feature space. In separate principal component analyses, ratio-derived variables showed a more concentrated low-dimensional covariance structure than absolute lipid concentrations. Circulating sphingolipids showed limited correspondence with tumor-local remodeling, whereas oxidative stress markers showed strong apparent discrimination between CRC and hospitalized non-cancer controls, although this finding may be affected by residual confounding. TCGA–GTEx analyses provided complementary pathway-level transcriptomic context, while anatomically resolved analysis of 374 TCGA tumors identified 3376 genes significantly associated with colorectal anatomical position, including six sphingolipid-related genes. Overall, these exploratory findings support a conceptual CRC Metabolic Continuum while requiring validation in larger, independent cohorts.

Antioxidants doi: 10.3390/antiox15091135

Authors: Irfan Zidni Abinawanto Abinawanto Akhmad Taufiq Mukti Aisyah Aisyah Suci Lestari Yuli Andriani Iskandar Iskandar Roffi Grandiosa Atikah Nurhayati Pringgo Kusuma Dwi Noor Yadi Putra Mochammad Ikhsan Cahya Utama Elva Yenizar Anggraeni

The freeze–thaw process in sperm cryopreservation generates reactive oxygen species (ROS) that impair motility, viability, and fertilizing capacity in fish, including the Javaen barb (Systomus rubripinnis), an endemic Indonesian cyprinid of conservation concern. Cryopreservation underpins gene bank development for aquatic genetic resources and supports biodiversity conservation alongside hatchery-based stock enhancement. This study evaluated five natural antioxidants: basil leaf extract (BLE), noni fruit extract (NFE), moringa leaf extract (MLE), sambiloto leaf extract (SLE), and honey–egg yolk (HEY). Each antioxidant was supplemented into the Kurokura’s cryopreservation extender across a graded concentration series (BLE 1–4%, NFE and MLE 0.5–1.5%, SLE 6–9%, and HEY 1–2% with a fixed 5% egg yolk) and compared with an antioxidant-free control. Radical scavenging capacity was measured by DPPH assay (IC50: BLE 134.06, MLE 297.04, NFE 422.86, SLE 3010.46 ppm), and post-thaw motility, duration of motility, viability, abnormality, fertilization rate, and ATP bioluminescence were assessed. The most effective doses tested were doses of 1% for BLE, NFE, and MLE, 1.25% for HEY, and 7% for SLE. BLE at 1% gave the best overall profile (motility 85.00 ± 0.88%; duration 7.14 ± 0.38 min; viability 82.00 ± 1.86%; abnormality 18.26 ± 0.16%), significantly exceeding the control (65.00 ± 2.31%; 4.56 ± 0.33 min; 68.00 ± 2.42%; 23.54 ± 0.28%; p < 0.05), whereas SLE sustained the longest motility. All extracts supported fertilization rates above 82% versus 70.08 ± 3.82% in the control, and 1% BLE preserved post-thaw luminescence equivalent to fresh sperm (165,217 vs. 164,087 RLU), interpreted comparatively. These locally available, plant-derived antioxidants offer a low-cost route to germplasm conservation of Javaen barb, strengthening conservation hatchery and broodstock management, sustainable aquaculture production, and food security for inland fisheries communities.

Antioxidants doi: 10.3390/antiox15091134

Authors: Ying Wen Pengfei Zhang Xinyu Liao Jiankang Liu Yang Zhang Xuyun Liu

Preservation of mitochondrial integrity has emerged as a central hub in the anti-inflammatory effect of exercise. This narrative review advances a framework in which mitochondrial damage-associated molecular patterns (mtDAMPs) serve as the mechanistic bridge between exercise and inflammation. Mitochondrial dysfunction releases mtDAMPs, including mitochondrial DNA (mtDNA), reactive oxygen species, cardiolipin, N-formyl peptides, and ATP, which activate cGAS–STING, the NLRP3 inflammasome, TLR9, AIM2, ZBP1, and NF-κB signaling. Crosstalk among these pathways allows mild mitochondrial damage to escalate into chronic inflammation. Exercise opposes this cascade through the AMPK–PGC-1α axis, which coordinately activates four mitochondrial quality control (MQC) modules: biogenesis, antioxidant defense, dynamics, and mitophagy. The cardiovascular system illustrates this framework, as myocardial inflammation runs mainly through mtDNA–cGAS–STING signaling and vascular inflammation through oxidized mtDNA–NLRP3 signaling, while cardiovascular aging engages both axes at once. Throughout, exercise refers to repeated training rather than to a single bout, and the framework targets middle-aged and older adults with, or at risk of, cardiovascular disease. The upstream half of the sequence, in which training raises mitochondrial content and antioxidant capacity, rests on human muscle biopsy data; the downstream half remains largely preclinical. MQC is therefore proposed as a testable target rather than an established one.

Antioxidants doi: 10.3390/antiox15091133

Authors: Joanna Kuć Mateusz Maciejczyk Krzysztof Dariusz Szarejko Małgorzata Żendzian-Piotrowska Walery Tarnawski Sara Zięba Anna Zalewska

Background: Temporomandibular disorders constitute multifaced systemic impairments capable of compromising overall physiological homeostasis. This study compares oxidative and nitrosative stress biomarkers between patients with temporomandibular myofascial pain with referral and healthy controls. Methods: We enrolled 44 individuals with temporomandibular myofascial pain with referrals and 44 controls. The procedure involved clinical examination based on the Diagnostic Criteria for Temporomandibular Disorders and saliva collection. Biochemical determination involved advanced glycation end products (AGEs), advanced oxidation protein products (AOPPs), thiobarbituric-acid-reactive substances (TBARS), and total nitric oxide (NO). Results: Median concentrations of TBARS and total NO were significantly elevated in the study group relative to the controls (TBARS: 6.078 vs. 1.486; total NO: 138.6 vs. 23.82), whereas the concentration of AOPPs was significantly lower (151.1 vs. 505.3). Multiple regression revealed that myofascial pain was an independent predictor of the observed differences, even after we adjusted for age and salivary flow rate (TBARS: β = 4.78 [95% CI: 2.020, 7.541], adjp = 0.0018; total NO β = 128.4 [95% CI: 82.70, 174.1] adjp = 0.0004; AOPP β = −755.7 [95% CI: −1228, −283.5] adjp-value = 0.0028. Conclusions: Myofascial pain with referral is independently associated with oxidative imbalance, highlighting the relevance of TBARS and total NO as promising severity biomarkers. The unexpected decrease in AGE and AOPP levels requires standardized studies to confirm diagnostic utility.

Antioxidants doi: 10.3390/antiox15091132

Authors: Shan Dou Bo Liu Malik Ahsan Ali Anqi Huang Changjun Zeng

Boar sperm are highly sensitive to cryopreservation, limiting the use of frozen semen in pig breeding. We previously reported that cryopreservation alters lysine acetylation of metabolism-related proteins, including the sperm-specific isoform LDHC. Here we show that cryopreservation reduces LDH activity, and that the LDH inhibitor galloflavin recapitulates this loss, confirming that LDHC is required for boar sperm motility, acrosome integrity, and mitochondrial membrane potential. Treatment with the SIRT inhibitor nicotinamide increased LDHC acetylation and simultaneously suppressed LDH activity, indicating that hyperacetylation negatively regulates LDHC function. Untargeted metabolomics of LDH-inhibited sperm revealed a notable decrease in guanidinoacetic acid (GAA), the sole precursor of creatine. Supplementing the freezing extender with 600 μM GAA significantly improved post-thaw motility, preserved mitochondrial membrane potential and ATP levels, reduced oxidative stress, and protected acrosome integrity. These improvements were accompanied by elevated intracellular creatine content, enhanced antioxidant capacity (increased T-AOC, CAT, and SOD activities, decreased MDA), and reduced apoptosis. Our findings demonstrate that cryopreservation-induced LDHC hyperacetylation impairs enzyme activity and sperm quality, and that GAA replenishment effectively counteracts this injury through creatine-dependent and antioxidant mechanisms.

Antioxidants doi: 10.3390/antiox15091130

Authors: Xuanjun Wang Jun Zhang Saizhen Guo Wenbo Zhang Ziyan Yang Dengyou Nie Zemin Xiang Yongkai Xi

As a traditional Tibetan medicine and health food, the fleshy roots of Sphallerocarpus gracilis were adopted to extract polyphenol powder(PP) in this research. Using a DSS-induced colitis mouse model combined with multi-omics analysis and experimental validation, we explored the protective effects and molecular mechanisms of PP against intestinal inflammation. PP mitigated colitis-related symptoms in a dose-dependent manner, inhibited pro-inflammatory cytokines and dose-dependently reversed elevated IL-10 levels to BC group. Multi-omics data revealed that PP intervention was associated with gut microbiota remodeling, enrichment of Akkermansia muciniphila, restoration of bacterial–fungal homeostasis, and improvement of tryptophan and biotin metabolism. By suppressing HMGB1 and RAGE expression, PP blocked the AGE-RAGE-mediated inflammatory cascade. In addition, PP maintained intestinal mucosal integrity through goblet cell protection, MDA reduction, and the regulation of BAX/Bcl-2, MMP3 and MMP9. Overall, PP relieves UC by regulating gut microecology, host metabolism and the AGE-RAGE pathway, which supports the translational potential of natural polyphenols for inflammatory bowel disease treatment as a gut microecological modulator.

Antioxidants doi: 10.3390/antiox15091131

Authors: Lorenzo Loffredo

Tobacco smoking is an established cardiovascular risk factor and is closely associated with the development and progression of atherosclerosis, as well as complications such as myocardial infarction and stroke [...]

Antioxidants doi: 10.3390/antiox15091129

Authors: Szymon Sip Anna Gościniak Niklas Hoede Min Hein Htike Hnin Yuzana Khin Wei-Wei Lai Hein Htet Naing Agnieszka Szopa Judyta Cielecka-Piontek

Flavonoids are a diverse group of naturally occurring polyphenolic compounds found in plant-based foods and are known for their broad-spectrum biological activities. Among them, nobiletin, a polymethoxylated flavone derived from citrus peels, has gained attention for its multifaceted role in promoting healthy ageing and mitigating age-related diseases. This review explores the chemical properties, mechanisms of action, therapeutic potential, and challenges related to nobiletin. The focus is placed on its antioxidant, anti-inflammatory, neuroprotective, cardioprotective, and metabolic regulatory effects, as well as its capacity to enhance mitophagy, modulate circadian rhythms, and prevent ferroptosis. Despite promising preclinical results across various models, including in vitro cell lines, C. elegans, and mice, the clinical translation of nobiletin is hindered by poor oral bioavailability, complex extraction, and limited human data. Novel delivery systems, such as nanoemulsions and phospholipid-based carriers, offer potential solutions to enhance its bioavailability. As research advances, nobiletin emerges as a promising candidate for geroprotective therapy, warranting further investigation through well-designed clinical trials to extend healthspan and prevent age-associated pathologies.

Antioxidants doi: 10.3390/antiox15091128

Authors: Ufuk Ülker Bülent Bayraktar Mehmet Eray Alçığır Sibel Kızıl Tünay Karan Gökşad Cemil Kotan Mustafa Yeni

Moringa oleifera has anti-inflammatory and antioxidant properties, but its effects in polymicrobial sepsis are incompletely defined. We evaluated M. oleifera oil (MOO) in female Wistar rats subjected to cecal ligation and puncture (CLP). Forty rats were randomized to Control, Sham, CLP, CLP + MOO 100 mg/kg, or CLP + MOO 200 mg/kg (n = 8/group). At 24 h, survival, clinical scores, serum apelin, omentin-1, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), malondialdehyde (MDA), recoverable bacterial burden, and histopathology were assessed. CLP decreased omentin-1 and increased IL-6, TNF-α, MDA, recoverable Staphylococcus aureus and Escherichia coli, clinical severity, and multi-organ injury. At 200 mg/kg, apelin and omentin-1 were 1.15 ± 0.12 and 28.4 ± 3.1 ng/mL, while IL-6, TNF-α, and MDA decreased to 18.4 ± 1.8 pg/mL, 24.1 ± 2.8 pg/mL, and 2.5 ± 0.3 nmol/mL, respectively. Neither organism was recovered above the detection limit. Survival was 62.5% in CLP, 87.5% with 100 mg/kg, and 100% with 200 mg/kg MOO. Within this 24 h model, a single post-CLP oral dose of MOO was associated with lower inflammatory cytokine concentrations and MDA, reduced recoverable bacterial burden, improved clinical and histopathological findings, and higher descriptive survival. Because MDA was the sole oxidative endpoint and no non-septic MOO-only group or molecular redox assays were included, these findings support the attenuation of sepsis-associated lipid peroxidation but do not establish a direct antioxidant mechanism.

Antioxidants doi: 10.3390/antiox15091127

Authors: Pedro Amorim Lívia de Sá Hayashide Vitor Emanuel Leocadio Mariana Marques Isabelle Navarra Cherley Borba Vieira Andrade Jorge José de Carvalho Rafael Serafim Pinto Luan Pereira Diniz

Astrocyte senescence is a recognized feature of brain aging, but its impact on neuronal mitochondrial homeostasis remains poorly defined, particularly in human cells. Here we show that doxorubicin-induced senescence disrupts mitochondrial function in primary human astrocytes and compromises their capacity to sustain neuronal bioenergetics. Senescent astrocytes accumulated a denser population of smaller, ultrastructurally damaged mitochondria together with increased levels of fission, fusion and biogenesis-associated proteins. Despite this apparent expansion of the mitochondrial compartment, these cells displayed reduced mitochondrial membrane potential, intracellular ATP and cellular metabolic activity, indicating accumulation of a functionally impaired mitochondrial population. Senescence also remodeled the extracellular mitochondrial compartment: conditioned medium from senescent astrocytes contained fewer mitochondrial particles with lower membrane potential and reduced ATP. Functionally, conditioned medium from control astrocytes increased TOMM20 and PGC-1α levels in human postmitotic neurons, whereas medium from senescent astrocytes failed to elicit this response and instead promoted hydrogen peroxide accumulation, ATP depletion and reduced cellular metabolic activity in the absence of overt cytotoxicity. Neurons acquired an astrocyte-derived MitoTracker signal from both conditions. Our data indicate that factors released by senescent human astrocytes are sufficient to induce neuronal mitochondrial and redox dysfunction.

Antioxidants doi: 10.3390/antiox15091126

Authors: Rifqah Azzahra Naulidia Michelle Ji Yeon Yoo

Natural deep eutectic solvents (NADES) offer a potentially food-compatible alternative for extracting plant polyphenols, although their direct application in yoghurt remains limited. This study evaluated the antioxidant and physicochemical properties of yoghurt fortified with SunGold kiwifruit extracts obtained using a selected choline chloride and glycerol NADES, with water extraction used for comparison. The phenolic profile of the NADES extract was characterised using LC-MS/MS. Both extracts were incorporated before (PRE) or after (POS) fermentation at concentrations of 10%, 20%, and 30% (v/v). The extracts and fortified yoghurts were analysed for total phenolic content (TPC), total flavonoid content (TFC), DPPH radical scavenging activity, FRAP, CUPRAC, and physicochemical properties. LC-MS/MS tentatively annotated several phenolic compounds in the NADES extract, with catechin showing the largest peak area among the annotated compounds. The water extract showed higher TPC and DPPH activity, whereas the NADES extract showed higher TFC, FRAP, and CUPRAC values. Increasing extract concentration generally increased the measured bioactive properties of the yoghurts, although the magnitude of change varied among assays and treatments. Water extract-fortified yoghurts generally showed higher TPC and DPPH values, while NADES extract-fortified yoghurts showed higher TFC, FRAP, and CUPRAC values. Extract type, fortification stage, and concentration also influenced pH, °Brix, viscosity, colour, and syneresis. Higher fortification concentrations reduced viscosity and increased syneresis, while low-concentration NADES extract-fortified yoghurts retained physicochemical properties closer to those of the control. Overall, the NADES extract showed potential as an ingredient for yoghurt fortification. Further studies are required to evaluate sensory acceptability, storage stability, starter culture viability, and gastrointestinal bioaccessibility.

Antioxidants doi: 10.3390/antiox15091125

Authors: Kuen-Jang Tsai Kuan-Tso Chen Chin-Chuan Tsai Zi-Xuan Hong Li-Ying Qiu Chan-Chuan Liu Kwang-Yu Chang Pin-Yuan Chen Chia-Hung Chien

Glioblastoma (GBM) frequently recurs after temozolomide (TMZ) therapy and exhibits substantial redox plasticity. Our previous work showed that the effects of hydrogen peroxide (H2O2) vary with its level and between parental and TMZ-resistant GBM cells. Succinate dehydrogenase subunit D (SDHD), a membrane-anchoring component of mitochondrial complex II, is positioned at the interface of electron transport and redox homeostasis, but its regulation in GBM remains unclear. We therefore examined whether SDHD expression changes across distinct H2O2-responsive contexts involving aquaporins (AQPs) and AKT. TCGA transcriptomic analysis showed higher SDHD mRNA expression in WHO grade IV than in grade II/III gliomas, whereas paired primary/recurrent high-grade glioma samples showed heterogeneous SDHD changes at recurrence. TMZ reduced SDHD, and SDHD knockdown decreased intracellular reactive oxygen species. Combined redox perturbation reduced SDHD, whereas AKT inhibition restored SDHD expression. Under receptor-associated signaling conditions, EGFRvIII expression or CXCL12 stimulation increased measured H2O2 together with AQP3, AKT Ser473 phosphorylation, and SDHD. In TMZ-resistant cells, pharmacological perturbation of aquaporin-associated signaling decreased AQP9 together with AKT Ser473 phosphorylation and SDHD. An AQP3-targeting compound further reduced cell density when combined with TMZ. Together, these findings suggest that changes in SDHD expression are consistent with a compensatory redox response in GBM cells and that its regulation varies with the nature of H2O2-associated signaling during TMZ-related stress.

Antioxidants doi: 10.3390/antiox15091124

Authors: Lorenzo Loffredo Enrico Maggio Simona Bartimoccia Vito Cantisani Antonio Angeloni Aurora Paraninfi Paolo Ciacci Simona Battaglia Federica Armeli Ilaria Maria Palumbo Mariaelena Malvasi Giancarlo D’Ambrosio Pasquale Pignatelli Roberto Carnevale Francesco Violi Francesco Barillà Gaetano Tanzilli

Takotsubo syndrome (TTS) is an acute and reversible heart failure condition characterized by transitional left ventricular systolic dysfunction without obstructive coronary artery disease. Although sympathetic hyperactivation is considered a key pathogenic mechanism, the contribution of gut-derived endotoxemia and oxidative stress is still unclear. Lipopolysaccharide (LPS), an endotoxin of Gram-negative bacteria, may translocate from the gut into the bloodstream and increase oxidative stress through activation of NADPH oxidase 2 (NOX2), nitric oxide (NO) depletion and endothelial dysfunction. This study aimed to evaluate circulating LPS levels in TTS and investigate their association with NOX2 activation, oxidative stress, and endothelial dysfunction. Twenty consecutive patients with TTS and 20 age- and sex-matched healthy controls were included. Within 48 h of admission, fasting blood samples were collected to assess soluble NOX2-derived peptide (sNOX2-dp), hydrogen peroxide (H2O2), NO metabolites (NOx), LPS, and zonulin. Endothelial function was assessed by brachial artery flow-mediated dilation (FMD). Compared with controls, TTS patients had significantly higher serum levels of sNOX2-dp, H2O2, LPS, and zonulin, lower NOx and impaired FMD. sNOX2-dp was positively correlated with LPS (Rs = 0.539, p < 0.001) and zonulin (Rs = 0.331, p = 0.037) and inversely correlated with FMD (Rs = −0.462, p = 0.003). NOx correlated negatively with H2O2, zonulin and LPS. In multivariable analysis, LPS was the only independent predictor of FMD (β = 0.498, SE = 0.126, p = 0.001) and sNOX2-dp (β = −0.572, SE = 0.034 p < 0.001); FMD (β = −0.347, SE = 0.455, p = 0.005), H2O2 (β = 0.445, SE = 0.155, p < 0.001), and zonulin (β = 0.298, SE = 2.309, p = 0.016) emerged as independent predictors of LPS (adjusted R2 = 0.585). TTS is associated with low-grade endotoxemia, NOX2-driven oxidative stress, reduced NO bioavailability, and endothelial dysfunction. The independent association between LPS and NOX2 activation supports a potential gut–vascular axis in TTS pathophysiology.

Antioxidants doi: 10.3390/antiox15091123

Authors: Meryem Kececi Oguzhanoglu Icten Olgu Bafali Kursat Oguzhanoglu Senem Karacabey Cakmak Busra Seker Atas Muhammed Oguz Yildiz Ali Cetin

Amniotic oxidative stress in women with vaginal infection but no documented intra-amniotic infection has received little attention. In this prospective cohort study of 90 women undergoing elective cesarean delivery, 45 had symptomatic, culture- or Nugent-confirmed vaginal infection and 45 were asymptomatic controls. Amniotic fluid advanced oxidation protein products (AOPP) and malondialdehyde (MDA) were measured by commercial ELISA. Both were higher in the infection group (median 7.96 versus 5.77 ng/mL and 12.49 versus 7.73 nmol/mL, both p < 0.001), with lower cord blood pH (p = 0.003) and more frequent neonatal intensive care unit (NICU) admission (26.7% versus 4.4%, p = 0.007). AOPP was associated with NICU admission (area under the curve 0.917, 95% CI 0.777 to 0.999), although this rests on 14 events with thresholds derived and evaluated in the same sample. Elevations were largest in the bacterial vaginosis and aerobic bacterial subgroups, but the etiologies did not differ. Both kits were designed for serum and are not validated for amniotic fluid, and 16.7% of MDA measurements fell outside the calibration range, so the MDA results are semi-quantitative. Vaginal infection at cesarean delivery is associated with higher amniotic oxidative stress markers; these findings are exploratory and require external validation.

Antioxidants doi: 10.3390/antiox15091122

Authors: Amonnat Sukhamwang Dumnoensun Pruksakorn Pornngarm Dejkriengkraikul Michael A. Dengler Supachai Yodkeeree

High-grade osteosarcoma presents a significant clinical challenge due to unpredictable therapeutic responses and aggressive progression. This study aimed to identify the critical molecular pathways driving chemotherapy resistance and aggressive phenotypes in osteosarcoma patients. Through transcriptomic and bioinformatic analyses, we identified cholesterol biosynthesis as a key upregulated metabolic pathway in poor chemotherapy responders, where squalene epoxidase (SQLE) emerged as an exploratory candidate hub gene whose elevated expression significantly correlates with shortened survival in the TCGA cohort. We validated these findings by administering terbinafine, a known SQLE inhibitor. In highly chemoresistant SaOS-2 cells exhibiting the highest baseline SQLE expression, terbinafine synergistically sensitized cells to doxorubicin by driving cell death partly through apoptosis, as confirmed by caspase inhibition. The combination also promoted ferroptosis, indicated by elevated ROS and MDA along with decreased FSP1 and GPX4 expression. Furthermore, the co-treatment effectively suppressed clonogenic potential, induced G2/M phase cell cycle arrest, and inhibited metastatic progression. These effects were mediated by the modulation of cell proliferation, metastasis, and survival genes through the coordinated regulation of the PI3K/AKT/mTOR, ERK, and JNK signaling cascades. Together, these results highlight the therapeutic potential of targeting the SQLE pathway to overcome doxorubicin resistance and suppress aggressive progression in high-grade osteosarcoma.

Antioxidants doi: 10.3390/antiox15091121

Authors: Jung Min Park Sohae Park Dae Youn Hwang Heeseob Lee Jumin Park

Porphyromonas gingivalis (Pg) is a keystone periodontal pathogen associated with biofilm formation and gingipain-mediated virulence. This study evaluated the antioxidant, antibacterial, antibiofilm, and antivirulence activities of a hexane extract of Lycopus lucidus (LLH) and its eight chromatographic fractions (H1–H8). Antioxidant activity was assessed using DPPH and ABTS radical-scavenging assays, whereas antibacterial activity, biofilm formation, and virulence-associated gene expression were evaluated using corresponding in vitro assays. LLH exhibited antioxidant and antibacterial activities, while H4 showed the strongest overall biological activity among the fractions. The DPPH IC50 values of LLH and H4 were 98.33 ± 2.05 and 60.67 ± 3.09 µg/mL, respectively, and the corresponding ABTS IC50 values were 89.24 ± 1.67 and 28.15 ± 1.21 µg/mL, respectively. H4 also showed greater inhibition of biofilm formation than LLH and more pronounced downregulation of several virulence-associated genes. LC–MS/MS analysis tentatively identified α-cyperone as a constituent of H4. Overall, chromatographic fractionation of LLH yielded H4 with enhanced biological activity across several measured endpoints, including radical-scavenging, antibacterial, and antibiofilm effects, together with more pronounced suppression of several virulence-associated genes. However, the contribution of α-cyperone or other individual constituents to these effects remains to be established.

Antioxidants doi: 10.3390/antiox15091120

Authors: Wenjing Wang Yuanbo Liu Jipeng Song Zouzou Yu Zixiang Chen Hu Jiao

Keloids are characterized by fibrosis and chronic inflammation, but links between mitochondrial dysfunction and keloid pathogenesis remain unclear. This study examined whether impaired PINK1/Parkin-dependent mitophagy is associated with mitochondrial DNA (mtDNA)-mediated innate immune activation and fibrosis in keloids, and evaluated mitochondrial transplantation as a potential therapeutic strategy. Primary keloid fibroblasts (KFs), normal skin fibroblasts (NFs), adipose-derived stem cells (ADSCs), human keloid tissues, and human keloid xenografts in immunodeficient BALB/c nude mice were analyzed using ultrastructural, molecular, and functional approaches. Freshly isolated NF-derived mitochondria (nMito) and ADSC-derived mitochondria (aMito) were compared at protein-equivalent doses. KFs exhibited mitochondrial abnormalities, impaired oxidative phosphorylation, increased reactive oxygen species, mtDNA leakage, and cGAS/STING pathway activation. Elevated PINK1 expression, reduced Parkin expression and p62 accumulation were consistent with impaired downstream mitophagic clearance. Both nMito and aMito were associated with improved mitochondrial function, changes in mitophagy-related markers, reduced cytosolic mtDNA and cGAS/STING signaling, and attenuated fibroblast activation, with greater aMito-associated changes in selected endpoints. In xenografts, intralesional administration of either mitochondria improved collagen organization and reduced fibrotic and inflammatory signaling. Together, these findings link altered PINK1/Parkin-dependent mitophagy to mtDNA-driven inflammation and fibrosis and support mitochondrial transplantation as a potential organelle-based therapeutic approach.

Antioxidants doi: 10.3390/antiox15091119

Authors: Sineenad Teerapatpaisan Alisa Naladta Kamonpan Wanichsri Penpimol Ponchunchoovong Suthasinee Thapphasaraphong Natsajee Nualkaew

Mung bean seed coat (MBSC) is a byproduct of commercial vermicelli production that possesses essential activities capable of delaying diabetic wound progression. This study aimed to evaluate the potential of MBSC extracts to promote diabetic wound healing, an effect that has not yet been reported. The hypoglycemic effect was assessed by glucose uptake stimulation in L6 myotubes and by an α-glucosidase inhibition assay. Antiglycation was determined by BSA-glucose and BSA-methylglyoxal assays. Intracellular reactive oxygen species (ROS) reduction and wound healing were assessed in human keratinocytes (HaCaT) exposed to high glucose (HG), and gene expression in HG-wounded cells was analyzed by qPCR. The results demonstrated that the ethanolic extract (EE) from MBSC exhibited glucose-lowering effects and suppressed glycation reactions at the early and intermediate stages, with IC50 values of 75 and 140 µg/mL, respectively. EE reduced ROS by 70%, stimulated cell proliferation by 57% in the high-glucose (HG)-exposed HaCaT cells, and accelerated cell migration to close the HG-exposed wound. EE increased the gene expression of Nrf2, NQO-1, SOD2, and CAT. It also downregulated TNF-α, upregulated TGF-β1, and downregulated MMP-9. In conclusion, EE has the potential to delay the progression of diabetic wounds by lowering blood glucose levels, inhibiting AGE and ROS formation, and enhancing cell proliferation and migration in HG-exposed HaCaT cells. The gene regulatory effects of EE were demonstrated as an Nrf2 activator that reduced oxidative stress, exerted anti-inflammatory effects, and regulated ECM balance. The preparation of oral and topical products could be further developed.

Antioxidants doi: 10.3390/antiox15091118

Authors: Edyta Gołaś Mateusz Maciejczyk

Skin aging is a complex process influenced by oxidative stress, protein glycation, chronic inflammation, and increased extracellular matrix remodeling. Intensive research is underway on new anti-aging substances with multi-target mechanisms of action while maintaining safety and efficacy. Ergothioneine, selenoneine, and ovothiol A are natural histidine derivatives of marine origin, in which the oxygen atom of the hydroxyl group has been replaced with sulphur or selenium. In recent years, a broad spectrum of their biological activity has been demonstrated. Despite the well-documented antioxidant potential of these compounds, their anti-aging effects, particularly in terms of antiglycation and anti-inflammatory activity, remain insufficiently understood. This study presents the current state of knowledge regarding the biological activity of ergothioneine, selenoneine, and ovothiol A, and discusses available cosmetic preparations and dietary supplements containing these compounds. Meanwhile, significant research gaps have been identified regarding their potential use in the prevention and treatment of skin aging.

Antioxidants doi: 10.3390/antiox15091117

Authors: Hellen P. Valerio Valeria Oliveira Stephanie Y. Ferreira Isabel R. Pereira Giuseppe Palmisano Mariana P. Massafera Vanderson S. Bispo Fernanda M. Prado Paolo Di Mascio Marisa H. G. Medeiros

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by progressive motor neuron degeneration and chronic neuroinflammation in the brain and spinal cord, involving complex interactions between neurons and immune cells. Carnosine (β-alanyl-L-histidine) has pathophysiological relevance due to its ability to detoxify reactive carbonyl species, including α,β-unsaturated aldehydes, scavenge free radicals, and chelate zinc, and has also been proposed to function in the central nervous system as a histidine reservoir for histamine synthesis. Here, we investigated the effects of carnosine supplementation on the cerebellar proteome of SOD1G93A ALS rats using quantitative proteomics. Carnosine treatment extensively remodeled mitochondrial, antioxidant, and synaptic vesicle-trafficking protein networks and increased the abundance of glutamatergic and GABAergic receptor subunits relative to untreated ALS animals, with several of these changes exceeding wild-type levels. Pathway enrichment analyses identified significant up-regulation of Rab-mediated vesicle trafficking, synaptic vesicle cycling, and neurotransmitter transport/secretion pathways, alongside a partial reduction in RNA splicing and proteasomal subunits that were elevated in untreated ALS animals. Cross-comparison with the ALS-associated proteomic signature revealed that most carnosine-responsive proteins followed, rather than reversed, the direction of disease-associated change, indicating that carnosine predominantly potentiates an endogenous compensatory program rather than restoring a wild-type-like proteome. Collectively, these findings show that carnosine drives systems-level remodeling of mitochondrial and synaptic networks in the ALS cerebellum, identifying candidate compensatory pathways and supporting further functional validation of carnosine as a component of multimodal therapeutic strategies in ALS.

Antioxidants doi: 10.3390/antiox15091115

Authors: Guodong Wang Jiawen Duan Longfei Sun Tao Xu Jianwei Chen Aihao Xu Quanhui Liu Mengqin Qin Shouyu Huo Weiqing Li Xiaozhen Li Quanqing Zou Prasanna Kallingappa Dandan Zhang Ben Huang

Background: Postpartum hypogalactia is a prevalent obstetric complication worldwide, closely associated with excessive oxidative stress and impaired antioxidant defense in mammary tissue. Current hormone-based therapies carry endocrine disruption risks, while natural antioxidant bioactive agents such as extracellular vesicles (EVs) are largely limited by rapid in vivo clearance and poor tissue retention. Methods: We constructed an injectable gelatin methacryloyl (GelMA) hydrogel system to encapsulate chemically induced mammary epithelial cell-derived EVs (CiMECs-EVs) and systematically evaluated their antioxidant and lactogenic activities via multi-omics analysis, cellular functional assays and a bromocriptine-induced murine hypogalactia model. Results: CiMECs-EVs induced a functional mammary epithelial-like phenotype in fibroblasts in a dose-dependent manner with functional cargo enriched in glutathione metabolism and redox-regulatory miRNAs. The GelMA matrix protected EV integrity and enabled sustained release, and the composite system significantly ameliorated mammary duct structure and lactation function in vivo with specific mammary tropism and no systemic toxicity, outperforming free EV treatment. Conclusions: This study presents a safe protein biomacromolecule-based antioxidant delivery platform that effectively restores mammary redox balance and antioxidant defenses, providing a promising non-hormonal therapeutic strategy for postpartum hypogalactia.

Antioxidants doi: 10.3390/antiox15091116

Authors: Federica De Luca Dario Troise Valentina Camporeale Giorgia Leccese Federica Galloso Roberto Cuttano Barbara Infante Giovanni Stallone Elena Ranieri Giuseppe Stefano Netti

Chronic kidney disease (CKD) represents a major global health challenge, affecting more than 10% of the population and contributing substantially to morbidity and premature mortality. Growing evidence identifies oxidative stress and mitochondrial dysfunction as central drivers of renal injury and disease progression across diverse etiologies. The kidney is one of the most mitochondria-rich organs in the body, reflecting the high bioenergetic demands required for tubular reabsorption and metabolic homeostasis. Disruption of mitochondrial oxidative phosphorylation, excessive production of reactive oxygen species (ROS), and impaired mitochondrial quality control mechanisms promote tubular injury, inflammation, and fibrosis. In particular, dysfunction of the electron transport chain, activation of NADPH oxidase isoforms—especially NOX4—and alterations in mitochondrial dynamics create a vicious cycle of oxidative damage and bioenergetic failure. Emerging evidence highlights the importance of mitochondrial quality control pathways, including fusion–fission balance, PINK1/Parkin-mediated mitophagy, and mitochondrial biogenesis regulated by PGC-1α and TFAM. Additional mechanisms include ferroptosis, epigenetic regulation, mitochondrial DNA-mediated innate immune activation, and Na+/K+-ATPase-linked redox signaling. At the translational level, redox and mitochondrial biomarkers and targeted therapies are biologically compelling, but the evidence is uneven: most candidate biomarkers remain insufficiently standardized, and direct mitochondria-targeted interventions are supported predominantly by preclinical studies or small human proof-of-concept trials. This review therefore emphasizes not only mechanistic advances but also conflicting findings, model limitations, and the barriers that currently separate experimental efficacy from clinically meaningful CKD outcomes.

Antioxidants doi: 10.3390/antiox15091114

Authors: Wen-Xu Yao Yao-Dan Ma Shi-Yao Ding Jian Lu Hao-Lan Tang Zeng-Ming Yang

Successful embryo implantation requires intimate crosstalk between the blastocyst and the uterine epithelium within a defined window of receptivity. However, the metabolic signals that mediate this process in mammals remain poorly understood. In this study, pregnant mice, primary uterine cell culture and uterine epithelial organoids were used to examine the regulation and function of heme oxygenase-1 (HO-1) during mouse embryo implantation and decidualization. We demonstrate that embryo-derived lactic acid drives heme catabolism and regulates epithelial receptivity in mice through a hypoxia-inducible factor 1α (HIF1α) -heme oxygenase-1 (HO-1) signaling axis. Specifically, lactic acid stabilizes HIF1α to induce HO-1 expression in uterine epithelial cells by promoting von Hippel-Lindau (VHL) nucleolar sequestration and downregulating PHD2/3. Additionally, lactic acid suppresses the transcriptional repressor BACH1, further facilitating HO-1 induction. At physiological heme levels, HO-1-derived bilirubin promotes epithelial receptivity by increasing phosphorylated STAT3 (p-STAT3) and downregulating MUC1. A low dose of hemin promotes epithelial receptivity and decidualization, whereas a high dose of hemin suppresses these processes. Pharmacological inhibition of HO-1 in mice markedly reduces implantation sites, establishing the functional necessity of this pathway. However, when heme levels exceed the regulatory capacity of HO-1, epithelial dysfunction ensues, characterized by reduced p-STAT3 and elevated MUC1, which ultimately disrupts implantation. Consistent with this, chronic heme exposure by oral gavage in mice increases uterine heme levels and upregulates BACH1, thereby suppressing HO-1 and trapping the uterus in a non-receptive state, causing implantation failure. Our findings define a lactic acid-HIF1α-HO-1-heme metabolic checkpoint that couples glycolytic signaling to heme regulation and endometrial receptivity. Dysregulation of this checkpoint may contribute to implantation disorders associated with heme stress, providing mechanistic insights into heme stress-related uterine receptivity failure.

Antioxidants doi: 10.3390/antiox15091113

Authors: Julia Płatkiewicz Joanna Wróbel Magdalena Jeszka-Skowron Robert Frankowski Zuzanna Grześkowiak Beata Czarczyńska-Goślińska Anna Maria Jeszka Agnieszka Zgoła-Grześkowiak

Herbs and spices are traditionally added to food in cuisines around the world. Antioxidant activity and content of bioactive compounds were compared in oregano, sage, basil, rosemary, and herbal mixtures. After optimization of ultrasound-assisted extraction of the ethanol–water extracts, the content of reducing compounds of the herb extracts was tested using the Folin–Ciocalteu method, and antioxidant capacity was evaluated with ABTS (2,2’-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt) and DPPH (2,2-diphenyl-1-picrylhydrazyl radical) assays. Oregano showed the highest antioxidant activity in all tests used (2.6 mg of gallic acid equivalent (GAE) per mL in the Folin–Ciocalteu test, 4.6 mg Trolox/mL in the ABTS assay, and 3.3 mg Trolox/mL in the DPPH assay) while rosemary had the lowest antioxidant activity (1.2 mg GAE/mL (Folin–Ciocalteu), 1.5 mg Trolox/mL (ABTS), and 1.3 mg Trolox/mL (DPPH)). Apart from antioxidant properties, the content of bioactive compounds was determined with the use of high-performance liquid chromatography–tandem mass spectrometry (LC-MS/MS). It was found that in all tested Lamiaceae herbs and the herbal mixes, rosmarinic acid widely predominates as a major non-volatile phenolic constituent, and its content varies from 1121 µg/g in rosemary to 10,255 µg/g in herbes de Provence. High concentrations were also observed for quinic acid in both oregano and rosemary. Interestingly, the concentrations of rosmarinic acid in the group of herbs studied are positively correlated with the results obtained in the Folin–Ciocalteu, ABTS, and DPPH tests (Spearman’s correlation coefficient 0.7030, 0.6657, and 0.7188, respectively), whereas no such correlation is observed for quinic acid. Overall, the findings indicate that these herbs share a common hydroxycinnamate-based phytochemical framework but display clear species-specific differences reflecting their intrinsic metabolism. Furthermore, the concentration of 3-caffeoylquinic acid in the Sicilian herbs (1021 µg/g) was approximately 10 times higher compared to the samples of Dalmatian herbs, herbes de Provence, and pure herbs, which demonstrates the unique chemical composition of that mixture, including the presence of dried tomatoes and tarragon, which were not included in other tested herbal mixtures.

Antioxidants doi: 10.3390/antiox15091112

Authors: Evgeny Pislyagin Igor Manzhulo Irina Agafonova Anna Starinets Ekaterina Menchinskaya Ekaterina Chingizova Darya Tarbeeva Sergey Fedoreyev Dmitry Aminin

Current Parkinson’s disease (PD) therapies like Levodopa (L-DOPA) only provide symptomatic relief, highlighting the need for multi-target neuroprotective agents. This study investigates the mechanisms and preclinical effects of trans-vitisin B (tVB), an oligomeric stilbene, in PD models. In LPS-stimulated HMC3 and RAW 264.7 cells, tVB (0.1–10.0 µM) significantly suppressed reactive oxygen species (ROS), nitric oxide (NO), COX-2, and pro-inflammatory cytokines (IL-1β, TNF-α), while restoring HSP70 chaperone levels to normalize proteostasis. These findings were validated in vivo using C57BL/6 mice with rotenone-induced chronic PD. Administration of tVB attenuated motor deficits (Cylinder test) and reduced pathological freezing (Open Field) and working memory impairments (Y-maze) in this model, without inducing the dyskinesia-like side effects of L-DOPA treatment in rodents. Histologically, tVB mitigated the loss of dopaminergic neurons (TH+) in the substantia nigra, reduced microglial activation (IBA-1+) and neuronal NO synthase, and suppressed pathological phosphorylated Tau protein (p-TauSer202) accumulation. Unlike L-DOPA’s direct dopaminergic stimulation, tVB’s neuroprotective efficacy is mediated through multilevel regulation of key PD pathogenetic pathways, including neuroinflammation, oxidative stress, and impaired proteostasis.

Antioxidants doi: 10.3390/antiox15091111

Authors: Caizhen Wang Ying Wang Hongyu Chen Bing Li Youran Shao Gen Zou

Oxidative stress drives skin aging, barrier impairment, and inflammatory amplification, making dietary antioxidants potential systemic contributors to cutaneous redox homeostasis. Edible fungi are distinctive sources of redox-active metabolites, particularly ergothioneine, a stable sulfur-containing antioxidant whose cellular uptake is mediated by the ergothioneine transporter OCTN1 (SLC22A4). This review evaluates ergothioneine, polysaccharides and β-glucans, cordycepin, phenolics, and Ganoderma triterpenoids as processing-sensitive dietary bioactives with redox relevance rather than topical cosmetic ingredients. We examine how cultivation, drying, cooking, extraction, fermentation, and microbial biomanufacturing determine antioxidant formation, retention, oral bioaccessibility, and dose realism, and how antioxidant response, inflammatory, mitochondrial, and gut microbiota-mediated pathways connect intake to skin endpoints. The strongest oral evidence concerns biomarker-linked ergothioneine-rich Pleurotus. Smaller Flammulina velutipes and Sparassis crispa trials report hydration or transepidermal water loss signals without comparable exposure biomarkers, whereas purified ergothioneine provides provisional non-mushroom food evidence. Compared with better-established oral ingredients, edible fungi offer distinctive food technology advantages but a narrower human evidence base. Priorities include processing-aware quality and safety markers, contaminant control, standardized digestion models, dose-realistic exposure estimates, and biomarker-anchored randomized human trials.

Antioxidants doi: 10.3390/antiox15091110

Authors: Rossella Grimaldi Francesca Franco Enzo Maria Vingolo

Retinitis pigmentosa (RP) is the most common inherited retinal dystrophy (prevalence ~1:4000) and a leading Mendelian cause of working-age blindness. Despite marked genetic heterogeneity, its progression converges on a common secondary cascade of outer-retinal hyperoxia, increased reactive oxygen species (ROS), and mitochondrial dysfunction that drives cone degeneration and central vision loss. Because this oxidative cascade is largely genotype-independent and pharmacologically tractable, oxidative stress is a cross-cutting therapeutic target. Within it, mitochondrial DNA (mtDNA) is a key element: once released from damaged photoreceptors—free or within exosomes—it may act as a damage-associated molecular pattern (DAMP), engaging TLR9, cGAS–STING, and the NLRP3 inflammasome and sustaining chronic neuroinflammation. Extracellular mtDNA is therefore a potential integrative marker, simultaneously reflecting oxidative stress, mitochondrial dysfunction, cell death, and innate-immune activation. A central knowledge gap, however, remains: the mechanistic steps linking mtDNA to inflammation and to photoreceptor death have not been demonstrated in RP itself, and extracellular mtDNA has never been quantified in the ocular fluids of RP patients. In this review we appraise oxidative biomarkers in RP, propose extracellular mtDNA as a candidate biomarker of disease activity, and examine antioxidant and redox-modulating therapies—from N-acetylcysteine and elamipretide trials to DAMP-sensor inhibition—across experimental and clinical models. Finally, we propose extracellular mtDNA as a candidate pharmacodynamic endpoint and outline a path toward its validation.

Antioxidants doi: 10.3390/antiox15091109

Authors: Francesca Miceli Eugenio Caradonna Claudia Panzano Wassim Mansour Fulvio Ferrara Lucy Costantino Carlo Setacci Luca di Marzo

Background: Trimethylamine N-oxide (TMAO) arises from the interaction of diet, gut microbial metabolism, hepatic oxidation, and renal clearance. Experimental work links TMAO exposure to mitochondrial oxidative stress, NLRP3 inflammasome activation, impaired nitric oxide signaling, vascular smooth muscle cell dysfunction, and thrombosis. How far these findings explain human vascular disease remains uncertain. Purpose: We examine TMAO and related metabolites in carotid atherosclerosis, aortic disease (abdominal aortic aneurysm, AAA, and dissection), and peripheral artery disease (PAD), focusing on redox biology and the obstacles that still limit clinical translation. Position: Current evidence makes the pathway biologically credible, but it does not support routine TMAO measurement, a universal cutoff, or treatment decisions based on a single metabolite. The recent association between γ-butyrobetaine and limb outcomes also suggests that TMAO may not always be the most informative component of the pathway. Most causal evidence remains preclinical, and no TMAO-lowering or redox-directed intervention has improved a vascular clinical endpoint. Conclusions: For now, the TMAO pathway remains investigational. Progress will depend on multicenter studies that measure several pathway metabolites with harmonized assays and carefully account for renal function, diet, and sex. Interventional studies are premature until safety and biological target engagement have been established.

Antioxidants doi: 10.3390/antiox15091108

Authors: Qiaoling Zhang Zhanqing Yang Xueyuan Yao Yinfei Xing Chenhao Wang Baiyu Li Zhanpeng Yue Bin Guo

As the only mammalian appendage capable of complete regeneration, deer antlers serve as an invaluable model to investigate cartilage regrowth, but the underlying mechanism remains unclear. This study revealed that addition of palmitic acid (PA), an abundant long-chain saturated free fatty acid, inhibited the proliferation and hypertrophy of antler chondrocytes while promoting chondrocyte apoptosis. PA treatment activated NOTCH1 signaling and restrained the transport of Ca2+ from the cytosol to the endoplasmic reticulum (ER) via RBPJ (recombination signal-binding protein for immunoglobulin kappa J region)-targeted TMTC4 (transmembrane O-mannosyltransferase targeting cadherins 4), resulting in a reduction in ER Ca2+. Meanwhile, PA disrupted the structure and function of mitochondria-associated ER membranes (MAMs) via TGM2 (transglutaminase 2) through the cytosolic Ca2+-mediated PPP3CB (protein phosphatase 3 catalytic subunit beta)-NFATC2 (nuclear factor of activated T cells cytoplasmic 2) pathway. Further analysis demonstrated that PA induced mitochondrial dysfunction via MAM-mediated mitochondrial Ca2+ insufficiency, thereby restricting mitophagy and attenuating lysosomal acidification. This caused the leakage of mitochondrial reactive oxygen species (mtROS) from depolarized mitochondria into the cytosol via the mitochondrial permeability transition pore, thereby inducing lipid peroxidation, while the addition of ROS scavengers prevented the negative effects of chondrocyte proliferation and hypertrophy and protected chondrocytes from apoptosis in the context of PA. Collectively, PA treatment regulated the proliferation, apoptosis and hypertrophy of antler chondrocytes by disrupting MAM function.

Antioxidants doi: 10.3390/antiox15091107

Authors: Milica D. Bogdanović Nina Devrnja Katarina B. Ćuković Janićijević Sofija Stupar Slađana I. Todorović Jelena Savić

Plants are continuously exposed to volatile organic compounds (VOCs) emitted by neighbors. Although the mechanisms governing VOC uptake and metabolism remain unclear, cytochrome P450 monooxygenases (CYP450s) are thought to participate in the detoxification and metabolic conversion of absorbed VOCs. Data from previously conducted cDNA microarray transcriptomic profiling in potato exposed to French marigold essential oil (FM-EO) was here used to filter differentially expressed sequences, and identified 54 unique CYP450 transcripts. Among the 10 most highly expressed sequences, two CYP81D1-like (81D1-1 and 81D1-2) and one CYP81D11-like (81D11-1) transcripts were found. RT-qPCR confirmed their strong induction within 8 h of volatile exposure. Comprehensive bioinformatics identified the most highly induced 81D1-1 gene as a CYP450 containing a predicted N-terminal hydrophobic signal or membrane-anchor region, the conserved heme-binding signature motif, and regulatory elements associated with oxidative stress responses. The other 81D11-1 gene, exhibiting a comparable expression level, was annotated only as a heme-binding protein but possessed seven distinct cis-regulatory elements, suggesting high transcriptional plasticity. Machine learning predictions assigned the highest interaction probability to (Z)-β-ocimene, whereas structure-based docking yielded the most favorable mean score for piperitone. This study provides the first characterization of the potato CYP450 superfamily in the context of volatile-mediated plant–plant interactions and identifies two CYP81D members as strong candidates for the oxidative metabolism of absorbed VOCs. The results support a proposed detoxification pathway in which CYP81-mediated oxidation precedes glutathione conjugation and intracellular sequestration of VOCs. These candidate genes provide a valuable foundation for future functional studies and may facilitate the development of sustainable crop protection strategies based on volatile-mediated plant defense.

Antioxidants doi: 10.3390/antiox15091106

Authors: Ochuko L. Erukainure Chika I. Chukwuma

Adipotoxicity is a major contributor to insulin resistance and type 2 diabetes, and increasing evidence highlights the gut–adipose axis as a promising therapeutic target. Soursop (Annona muricata) is rich in phytochemicals that can be biotransformed by the gut microbiota into bioactive metabolites with metabolic benefits. The present study investigated whether metabolites generated by in vitro fecal fermentation of soursop fruit (SWSF) and peel (SWSFP) protect against fructose-induced adipotoxicity. SWSF and SWSFP were fermented with rat fecal microbiota, and the resulting metabolites were evaluated in an ex vivo fructose-induced adipotoxicity model using perigonadal white adipose tissue. Activities of enzymes involved in glucose metabolism, the polyol pathway, glutathione metabolism, glyoxalase-1 activity, purinergic signaling, and inflammatory lipid metabolism were determined. GC–MS-based metabolomics and pathway enrichment analyses were performed on fecal and adipose tissues. Soursop fermentation significantly remodeled the fecal metabolome, enriching metabolites associated with fatty acid metabolism, glycerolipid metabolism, β-oxidation, sterol metabolism, and arachidonic acid metabolism. Fructose-induced adipotoxicity disrupted glucose metabolism, activated the polyol pathway, impaired glutathione metabolism and glyoxalase-1 activity, suppressed ATPase and ENTPDase activities, and elevated 5-LOX and 12/15-LOX activities. Treatment with soursop-enriched fecal metabolites significantly reversed these alterations in a dose-dependent manner. Adipose metabolomics further demonstrated restoration of pathways associated with fatty acid biosynthesis, mitochondrial β-oxidation, glycerolipid metabolism, steroid biosynthesis, and polyunsaturated fatty acid metabolism, indicating improved lipid homeostasis and reduced inflammatory lipid signaling. SWSF and SWSFP generally exhibited greater metabolic protection than the reference antioxidant compound, gallic acid. Soursop-derived gut metabolites attenuate fructose-induced adipotoxicity by coordinately restoring glucose metabolism, redox homeostasis, carbonyl detoxification, purinergic signaling, and lipid metabolism through the gut–adipose axis. These results suggest soursop as a potential functional food for preventing and managing adipose tissue dysfunction and metabolic disorders.

Antioxidants doi: 10.3390/antiox15091105

Authors: Venera Berisha-Muharremi Bernard Tahirbegolli Alberta Humolli Reem Hanna

Background: Chronic autoimmune thyroiditis (CAT) is characterized by persistent autoimmune activity and increased oxidative stress, which may contribute to thyroid follicular injury and disease progression. Photobiomodulation (PBM) has shown potential beneficial effects on thyroid function and autoimmunity; however, its independent effect on systemic antioxidant status, particularly serum glutathione (GSH), has not been established. This study aimed to evaluate the effect of thyroid-directed PBM on serum GSH concentrations and thyroid-related outcomes in treatment-naïve euthyroid women with CAT. Methods: This prospective, non-randomized, open-label, parallel-group comparative interventional study included 50 treatment-naïve euthyroid women with CAT. Participants were allocated to receive either thyroid-directed transdermal PBM (n = 25) or selenium plus vitamin D supplementation (n = 25). The primary outcome was the change in serum GSH concentration. Secondary outcomes included changes in thyroid-stimulating hormone (TSH), free triiodothyronine (FT3), free thyroxine (FT4), anti-thyroid peroxidase antibodies (anti-TPO), anti-thyroglobulin antibodies (anti-Tg), thyroid volume, and anthropometric parameters. Assessments were performed at baseline (T0), after the intervention period (T1), and three months after intervention completion (T2). Results: A significant time × group interaction was observed for serum GSH concentrations (F = 19.101, p < 0.0001). Significant interactions were also observed for anti-TPO (F = 7.513, p = 0.001), anti-Tg (F = 7.389, p = 0.002), and thyroid volume (F = 13.081, p < 0.0001). In the PBM group, GSH increased from baseline to T1 and remained higher at T2, while TSH, anti-TPO, anti-Tg, and thyroid volume decreased, and FT3 and FT4 increased. No significant longitudinal changes in GSH or thyroid autoantibodies were observed in the supplementation group. No participant in the PBM group required levothyroxine (LT4) initiation during follow-up, whereas 24% of participants in the supplementation group initiated LT4 therapy at T1, with some requiring dose escalation by T2. Conclusions: Thyroid-directed PBM was associated with improved systemic GSH status and favorable changes in thyroid autoimmunity, thyroid function parameters, and thyroid volume compared with selenium plus vitamin D supplementation in treatment-naïve euthyroid women with CAT. These findings suggest that modulation of oxidative stress may represent one potential biological pathway underlying the observed effects of PBM. Larger randomized controlled studies with longer follow-up are required to confirm these findings and define the clinical role of PBM in autoimmune thyroid disease.

Antioxidants doi: 10.3390/antiox15091104

Authors: Silvia Baldacci Elisa Bustaffa Olivia Curzio Andrea Borghini Gabriele Donzelli Chiara Cavigli Fabrizio Minichilli

Cardiovascular diseases remain the leading cause of global mortality, and environmental toxic metals and metalloids (TMMs) exposure has emerged as an important and potentially modifiable cardiovascular (CV) risk factor. Due to their persistence, bioaccumulation, and widespread distribution, represent a significant public health concern and have been associated with adverse CV outcomes. Mechanistically, TMM-induced cardiotoxicity is primarily mediated through oxidative stress (OS) and inflammatory pathways. Accordingly, related biomarkers, may be considered markers of biological effect and potential pathophysiological mediators associated with TMM exposure. This narrative review summarizes epidemiological evidence linking environmental TMMs exposure with OS and inflammatory biomarkers to CV outcomes. A structured literature search identified 1915 records in PubMed, of which 42 epidemiological studies met the inclusion criteria. The reviewed evidence consistently indicates that environmental and occupational TMMs exposure is associated with significant alterations in biomarkers of lipid peroxidation, antioxidant defense, systemic inflammation, and endothelial dysfunction, all of which reflect early biological CV effects associated with exposure. However, substantial heterogeneity across studies and the predominance of cross-sectional designs limit causal inference and the evaluation of their predictive capacity for future CV events. Future prospective studies integrating mixture-based exposure assessment, standardized biomarker measurements, and multi-omics approaches are needed to clarify whether these biomarkers can serve as markers of biological effect or subclinical CV toxicity and as potential mediators of CV risk, while also assessing their potential contribution to cardiovascular risk stratification in exposed populations.

Antioxidants doi: 10.3390/antiox15091103

Authors: Fei Su Liwen Liang Bin Yu Lihua Xu Hongchao Sun Shiyi Ye Xiufang Yuan Yin Xue Canying Liu Junxing Li

Shiga toxin-producing Escherichia coli (STEC) infection causes severe enteropathies and hemolytic uremic syndrome, yet non-antibiotic therapeutic strategies remain limited. This study evaluated whether gypenosides (GPs) protect against STEC-induced intestinal injury and investigated the underlying mechanisms. Mice were pretreated with GPs prior to STEC challenge, and protective effects were comprehensively assessed through survival analysis, bacterial burden quantification, transcriptomic profiling, flow cytometry, electron microscopy, and Western blotting. GPs significantly improved survival rates, alleviated diarrheal symptoms, reduced bacterial dissemination in systemic organs, and lowered Stx2 toxin levels. Furthermore, GPs effectively restored gut microbiota dysbiosis by enriching beneficial bacterial genera. Transcriptomic analysis suggested that GP treatment was associated with suppression of ferroptosis-related signaling pathways. Ultrastructural examination demonstrated ameliorated mitochondrial vacuolization, and biochemical assays confirmed decreased Fe2+ accumulation and attenuated lipid peroxidation. Mechanistically, these changes were associated with modulation of the ATF4-CHAC1-GPX4 axis and suppression of ferroptosis, together with preservation of mitochondrial integrity. Collectively, these findings identify GPs as a promising protective candidate against STEC-induced intestinal injury and highlight the ATF4-CHAC1-GPX4 axis as a candidate pathway for further mechanistic investigation.

Antioxidants doi: 10.3390/antiox15091102

Authors: Yiwen Yuan Zhenxian An Min Hu Xuebin Li Jiahao Wang Xuejing Liu Wenhao Zhang Zujie Xu Xiaoqin Zhao

High-fat diet (HFD)-induced skeletal muscle atrophy is characterized by impaired muscle mass and function, mitochondrial dysfunction, redox imbalance, and increased inflammation. Previous research has shown that aerobic exercise ameliorates HFD-induced skeletal muscle atrophy, but the underlying mechanisms remain unclear. Dynamin-related protein 1 (Drp1) is a key GTPase that mediates mitochondrial fission, maintains mitochondrial homeostasis, and regulates reactive oxygen species (ROS) production and inflammatory responses. This study investigated the potential involvement of Drp1-associated mitochondrial fission in the protective effects of aerobic exercise against HFD-induced skeletal muscle atrophy. HFD-fed mice underwent an 8-week aerobic exercise intervention, and Mdivi-1, a commonly used mitochondrial fission inhibitor, was administered intraperitoneally to further examine the involvement of mitochondrial fission. Assessments included grip strength, endurance testing, body composition, histology, transmission electron microscopy, immunofluorescence, DHE staining, antioxidant assays, Western blotting, and qPCR. Aerobic exercise reduced Drp1 phosphorylation, oxidative stress, and inflammatory responses in the skeletal muscle of HFD-fed mice and attenuated skeletal muscle atrophy. Mdivi-1 treatment produced similar protective effects, including attenuation of skeletal muscle atrophy, oxidative stress, and inflammatory responses. Together, these findings suggest that Drp1 phosphorylation and associated mitochondrial fission may contribute to the protective effects of aerobic exercise against HFD-induced skeletal muscle atrophy.

Antioxidants doi: 10.3390/antiox15091101

Authors: Wenkui Ma Qianru Chen Depeng Yin Pengyun Ji Liming Liu Xihe Li Bingyuan Wang Lu Zhang Guoshi Liu

Ovum pick-up (OPU) enables high-frequency oocyte retrieval from selected dairy cow donors, facilitating favorable genetic progression. However, oocytes obtained via OPU and subjected to in vitro maturation frequently suffer from severe oxidative stress, mitochondrial dysfunction, and DNA damage, resulting in lower pregnancy rates compared to those of in vivo-derived embryos. Melatonin (MT), as a potent antioxidant, may exert beneficial effects on these OPU-retrieved oocytes. In the current study, by combining in vitro and in vivo experiments, the protective effects and potential molecular mechanisms of MT on disrupted oocyte quality and development caused by OPU were systemically investigated. By integrating oocyte quality assessment, follicular fluid metabolomics, and blastocyst transcriptomics, the results showed that subcutaneous administration of MT to cows prior to OPU significantly increased the number of high-quality Grade A oocytes and reduced the number of low-quality Grade D oocytes compared to the control. These improvements were accompanied by reduced local and systemic oxidative stress, enhanced glutathione metabolism, and lower serum progesterone concentrations at the measured time points, coinciding with a more favorable endocrine environment for follicular development. The results from untargeted metabolomics of the follicular fluid revealed that MT supplementation was associated with an altered follicular fluid microenvironment characterized by the upregulation of estrogen derivatives and glutathione-related pathways. Transcriptomic analysis of blastocysts showed significant upregulation of the rate-limiting genes steroidogenic acute regulatory protein (STAR) and 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), suggesting a potential role in promoting the synthesis of reproductive hormones and maintaining the progress of embryo development. The in vitro study showed that MT supplementation at a concentration of 10−7 mol/L in oocyte maturation media significantly improved cleavage rates, blastocyst rates, and total blastocyst cell numbers compared to the control. In vitro evaluations demonstrated that MT scavenged intracellular reactive oxygen species (ROS), while transcriptomic analysis indicated a reprogramming of the embryonic transcriptome. These transcriptomic changes were associated with the AMPK, FoxO, and autophagy pathways, as well as the modulation of endoplasmic reticulum stress and cellular senescence. The results from both the in vivo and in vitro studies suggested that MT supplementation was associated with a follicular microenvironment favorable for oocyte growth and protected embryos from oxidative damage. Therefore, MT supplementation effectively reduced the disrupted oocyte development caused by OPU and improved oocyte recovery and quality in general. These findings provide promising experimental evidence for the potential application of MT in dairy breeding and offer insights into the associated molecular pathways. However, further functional validation is required to fully elucidate the underlying mechanisms and establish its large-scale applicability.

Antioxidants doi: 10.3390/antiox15091100

Authors: Qingman Yang Yuhong Liao Huihui Quan Qing Qiu Huahua Mao Dachun Gong Xuan Tu Zirui Wang Hao Fu Xionge Pi

The production performance, muscle quality, and gut microbiota of aquatic animals are closely associated with host growth stage and disease status. This study therefore compared the gut health, antioxidant capacity, and muscle nutritional composition of greenhouse-cultured Pelodiscus sinensis covering two growth stages (juvenile and adult) and two physiological states (healthy and diseased). The results showed that healthy turtles possessed significantly longer and denser intestinal villi compared with diseased turtles (p < 0.01), and adult turtles maintained higher activities of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and lysozyme (LZM) than juveniles regardless of health condition. Digestive enzyme profiles varied with age and health. Compared with adult turtles, juveniles exhibited significantly higher trypsin activity in both healthy and diseased groups (p < 0.001). Conversely, juveniles displayed significantly lower lipase activity than adults in the diseased group (p = 0.008). For muscle nutritional composition, adults contained higher crude protein (CP), essential amino acids (EAAs), and umami amino acids (UAAs) than juveniles, while healthy turtles had significantly better muscle quality indices than diseased individuals (p < 0.05). Meanwhile, gut microbial composition differed significantly between growth stages and health states (p = 0.001). The relative abundance of unclassified_f_Lachnospiraceae and Clostridium_sensu_stricto_2 was positively correlated with GSH-Px and muscle nutritional composition indicators (CP, EAAs, and UAAs), whereas it was negatively correlated with liver alkaline phosphatase (ALP), acid phosphatase (ACP), and intestinal enzyme activities. Data also showed that unclassified_o_Bacteroidales and unclassified_f_Lachnospiraceae may affect muscle nutritional composition and non-specific immunity by regulating the microbial functional pathways of amino acid metabolism, the digestive system, and non-specific immunity system. Collectively, these observations offer insights into variations in physiological traits of greenhouse-cultured P. sinensis. These correlational findings suggest that gut microbiota are closely associated with muscle nutrition and antioxidant status, providing valuable insights for sustainable soft-shelled turtle aquaculture.

Antioxidants doi: 10.3390/antiox15091099

Authors: Xin Xin Xiangzi Li Shiyu Jin Xiaoxu Huang Xu Gao Qiang Li Xin Jin

Oxidative stress disrupts skeletal muscle cell survival and proteostasis, thereby compromising animal skeletal muscle health. As an organic selenium source, selenomethionine (SeMet) participates in antioxidant defense and selenoprotein regulation. However, its protective role in hydrogen peroxide (H2O2)-induced injury in C2C12 myoblasts and its association with unfolded protein response (UPR)-related molecular changes remain incompletely understood. This study investigated whether SeMet alleviates H2O2-induced injury in C2C12 myoblasts and whether this effect is associated with UPR-related marker changes. C2C12 myoblasts were treated with H2O2 and SeMet, and cell survival was evaluated using cell viability assays and calcein acetoxymethyl ester/propidium iodide co-staining. The expression of selenoprotein-, UPR-, and apoptosis-related genes was examined by quantitative real-time PCR, and corresponding protein levels were assessed by Western blotting. SeMet improved cell survival under oxidative stress and increased Selenof mRNA and SELENOF protein expression. It also attenuated aberrant changes in endoplasmic reticulum and mitochondrial UPR-related markers and reduced apoptosis-associated molecular responses. These findings indicate that SeMet protects C2C12 myoblasts against H2O2-induced oxidative injury. This protective effect may be associated with changes in SELENOF expression, UPR-related markers, and apoptosis-associated molecular responses, providing molecular evidence for its potential role in maintaining skeletal muscle cell homeostasis.

Antioxidants doi: 10.3390/antiox15091098

Authors: Michał Radzikowski Mateusz Maciejczyk

Allergies and diabetes mellitus are highly prevalent, frequently co-occurring diseases. Antihistamines, the first-line treatment for allergies, block H1 receptors but also exhibit anti-inflammatory and antioxidant properties that may improve glucose metabolism. This systematic review examined the evidence for the potential antidiabetic effects of commonly used first- and second-generation antihistamines. A comprehensive search of the PubMed database (1978–May 2026) yielded 17 eligible in vivo, in vitro, clinical, and in silico studies. Most reports indicated that antihistamines improve glucose tolerance, insulin sensitivity, and insulin secretion. Proposed mechanisms include enhanced insulin release, mast cell stabilization, attenuation of immune-mediated responses, reduced nephron alteration, and improved incretin signalling. Although substantial heterogeneity in study designs limits direct comparisons, current evidence suggests a beneficial role of antihistamines in modulating glucose metabolism, particularly for patients managing both allergies and diabetes. These findings highlight the potential for drug repurposing. However, further well-designed clinical trials are required to ensure reproducibility, successful animal-to-human translation, and overall safety before clinical implementation.

Antioxidants doi: 10.3390/antiox15091097

Authors: Hassan Barakat Ahmed H. Bahloul Ahmed A. H. Abddelatif

This integrative review examines the potential of dates, omega-3 (ω-3) and omega-6 (ω-6) polyunsaturated fatty acids (PUFAs), dietary fibers (DFs), and probiotic bacteria to support gut health through complementary nutritional pathways. The literature was searched in PubMed, Web of Science, Scopus, and Google Scholar for English-language studies published through June 2026. Dates contain fermentable substrates, including DFs and non-digestible oligosaccharides, that can serve as growth substrates for beneficial gut microbes. However, the whole fruit has not been formally established as a prebiotic under current consensus definitions. ω-3 and ω-6 fatty acids contribute to membrane integrity, eicosanoid synthesis, immune regulation, and inflammatory signaling, with effects influenced by dietary balance. DFs support gastrointestinal function, glycemic regulation, lipid metabolism, and colonic fermentation, including short-chain fatty acid (SCFA) production, whereas probiotics may exert strain-specific effects on microbial balance, barrier function, immune responses, and host metabolism. Together, these components may influence gut microbiota, oxidative stress, inflammation, and metabolic homeostasis. However, the evidence is heterogeneous, combined interventions are scarce, and several proposed effects rely mainly on preclinical studies or surrogate human outcomes. Thus, potential collaborative effects and disease-prevention implications remain unconfirmed. Well-designed human trials using standardized preparations, realistic intake levels, and clinically relevant outcomes are needed to clarify their combined effects.

Antioxidants doi: 10.3390/antiox15091096

Authors: Juexi Liu Lingtong Fan Jingran Wei Qingsong Liu Ouyan Han Yan Yang Danjun Guo Wei Xu Huajuan Wang E Liao

Population aging has made sarcopenia a growing concern in geriatric health. Protein–polyphenol non-covalent complexes can serve as carrier systems that improve the stability and bioactivity of natural antioxidants. This study refined the preparation parameters for the non-covalent complex of whey protein isolate (WPI) and resveratrol (RES), achieving protein digestibility of 83.60 ± 0.50% and DPPH scavenging of 52.41 ± 0.58% at pH 7.0, a WPI:RES molar ratio of 1:1, and a reaction time of 1.5 h. Relative to free WPI, the complex improved DPPH scavenging by 21.66% while preserving protein digestibility. The binding mode, interaction forces, and conformational evolution were investigated via spectroscopic experiments, docking studies and molecular dynamics simulations. RES selectively bound to the surface of β-lactoglobulin in WPI through hydrophobic interactions and hydrogen bonding, with a docking score of −6.366 kcal/mol and an MM-GBSA binding free energy of −30.48 kcal/mol. The complex maintained a highly stable conformation throughout the 100 ns molecular dynamics simulation. In conclusion, this study elucidated the structural changes of WPI upon non-covalent resveratrol binding. The WPI-RES complex exhibited enhanced DPPH radical scavenging activity while preserving protein digestibility, with potential implications for functional food development in sarcopenia management.

Antioxidants doi: 10.3390/antiox15091093

Authors: Vinuka De Silva Lochlan J. Fennell Mitchell A. Sullivan

Diabetic retinopathy (DR) is a major cause of preventable visual impairment and develops through intertwined metabolic, oxidative, inflammatory, and neurovascular disturbances. This review examines how chronic hyperglycaemia disrupts retinal glucose handling and redox homeostasis, with particular emphasis on glycogen metabolism as an underappreciated contributor to disease progression. Pathological glycogen accumulation in retinal amacrine cells and the retinal pigment epithelium may arise through altered glycogen synthase localisation and glucose-6-phosphate-dependent activation, potentially disturbing intracellular trafficking and cellular energy balance. These metabolic changes converge with mitochondrial electron transport chain dysfunction, NADPH oxidase activation, polyol pathway flux, and light-driven lipid peroxidation to increase reactive oxygen species generation. At the same time, transient suppression of Nrf2-dependent antioxidant defences, TXNIP-NLRP3 inflammasome signalling, ferroptotic injury, and VEGF-associated oxidative feedback promote blood–retinal barrier breakdown and persistent neuroinflammation. We propose that dysregulated glycogen metabolism and impaired antioxidant capacity form an integrated metabolic–redox network that helps explain cell-specific vulnerability and metabolic memory in DR. Targeting multiple nodes within this network may support earlier, disease-modifying strategies beyond treatment of advanced vascular complications.

Antioxidants doi: 10.3390/antiox15091095

Authors: Adriana Albini Danilo Morelli Cecilia Bender Paola Corradino

Skin health is increasingly recognised as a key determinant of overall well-being, and its decline is driven by multiple interconnected biological processes. Hydroxytyrosol (HT), an olive polyphenol, exhibits pleiotropic biological activities and has a potential role in supporting skin homeostasis. This narrative review critically evaluates the evidence on oral HT and HT-rich olive-derived nutraceuticals for skin health and healthy ageing with a focus on oxidative damage, inflammation, photoprotection, extracellular matrix (ECM) preservation, wound healing, and skin pigmentation conditions. HT exhibits high oral bioavailability and modulates antioxidant defences, inflammatory signalling, autophagy, apoptosis, and metabolic homeostasis. Preclinical studies consistently show protective effects against oxidative stress, ECM degradation, cellular senescence, photoageing, impaired wound healing, and ultraviolet-induced skin damage. Experimental evidence also indicates antiproliferative and pro-apoptotic effects in melanoma models through modulation of oncogenic signalling pathways, suggesting a potential role in skin cancer prevention. In vivo, oral HT administration improves glycative skin ageing, psoriasis, intestinal barrier integrity, and systemic inflammation. Clinical evidence supports systemic antioxidant and cardiometabolic benefits of purified HT and HT-rich formulations, while dermatological evidence remains preliminary for psoriasis, melasma, and wound healing. Overall, HT shows promise, but randomised controlled trials with validated dermatological endpoints are needed to confirm its clinical efficacy.

Antioxidants doi: 10.3390/antiox15091094

Authors: Minha Jeong Juhyeong Seo Sunwoo Park Jiyeon Ham

Canine osteosarcoma is an aggressive primary bone malignancy associated with a poor prognosis and a high propensity for lung metastasis. Due to the limitations and side effects of conventional chemotherapy, there remains a pressing need for alternative therapeutic strategies and adjuvants. In this study, we examined the effects of berberine chloride (BBRC), a bioactive isoquinoline alkaloid derived from the dried roots of Coptidis Rhizoma, on canine osteosarcoma D-17 and DSN cell lines. BBRC reduced the viability and migratory capacity of D-17 and DSN cell lines in three-dimensional (3D) spheroid cultures. Additionally, BBRC induced calcium ion overload in the mitochondrial matrix but not in the cytoplasm, in both osteosarcoma cell lines. BBRC also triggered excessive generation of reactive oxygen species (ROS) in both cell lines. Furthermore, Western Blot analysis revealed that BBRC downregulated the MMP2/AKT/β-catenin signaling pathways in both BBRC in D-17 and DSN cells. In addition, BBRC downregulated Cyclin D1 and Survivin, known downstream targets of the β-catenin pathway. We also confirmed that BBRC suppressed the transcription of genes involved in cell cycle regulation. Overall, our findings indicate that BBRC inhibits osteosarcoma cell growth by modulating calcium-redox homeostasis and the MMP2/AKT/β-catenin pathway.

Antioxidants doi: 10.3390/antiox15091091

Authors: Elena Coyago-Cruz Gabriela Méndez Ruth Escobar-Quiñonez Gabriela Cisneros Jorge Heredia-Moya

This study evaluated changes in physicochemical characteristics, bioactive compounds, in vitro antioxidant activity, and antimicrobial activity during four ripening stages (M0%, M50%, M80%, and M100%) of ‘Castilla’ blackberry, ‘Buckingham Tayberry’ raspberry, ‘Heritage’ raspberry, blueberry, and ‘Mortiño’. Bioactive compounds were quantified using rapid-resolution liquid chromatography, antioxidant activity was determined using the ABTS and DPPH assays, and antimicrobial activity was evaluated through minimum inhibitory concentration assays against 13 bacteria and 7 fungal strains. Ripening effects were fruit- and compound-dependent. Soluble solids generally increased, and titratable acidity decreased during ripening. Early stages were characterised by higher concentrations of several organic and phenolic acids and, in some fruits, greater radical scavenging and antimicrobial responses. Advanced ripening favoured the accumulation of carotenoids and anthocyanins, particularly in blueberry and ‘Mortiño’. Antimicrobial activity also varies according to fruit type, ripening stage, and microorganism. ‘Mortiño’ showed the lowest MIC against Enterococcus faecalis, whereas the other fruits displayed selective inhibition against different bacterial and fungal strains. Overall, no single ripening stage maximised all compositional and biological variables, demonstrating that harvest stage selection should be fruit- and purpose-specific.

Antioxidants doi: 10.3390/antiox15091092

Authors: Guangquan Li Kaiqi Weng Yi Liu Xianze Wang Huiying Wang Daqian He

Clothianidin (CLO), a widely used neonicotinoid insecticide, may pose a health risk to waterfowl through contaminated feed ingredients and agricultural residues. This study investigated whether dietary Bacillus amyloliquefaciens (BA) could alleviate CLO-induced growth suppression and liver oxidative injury in goslings. Fourteen-day-old Holdobagy goslings were assigned to five treatments for 21 days: control, 2.5 mg/kg CLO, 2.5 mg/kg CLO plus 60 ppm BA, 10 mg/kg CLO, and 10 mg/kg CLO plus 60 ppm BA. CLO exposure reduced final body weight and average daily gain, increased liver index, elevated serum alkaline phosphatase (AKP), aspartate aminotransferase (AST), and malondialdehyde (MDA) levels, and decreased liver total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) activities (p < 0.05). Histological analysis further showed hepatocyte disorganization, vacuolar degeneration, congestion, and inflammatory infiltration, which were partially alleviated by BA supplementation. Serum untargeted metabolomics showed clear separation among treatment groups. CLO exposure altered 130 and 215 differential metabolites at the low and high doses, respectively, whereas BA supplementation regulated 115 and 170 differential metabolites under the corresponding CLO backgrounds (p < 0.05). These metabolites were mainly enriched in amino acid metabolism, ATP-binding cassette (ABC) transporters, purine metabolism, riboflavin metabolism, glutathione metabolism, glycerophospholipid metabolism, and fatty acid biosynthesis/degradation pathways. Liver transcriptomic analysis identified 761 and 1283 differentially expressed genes after low- and high-dose CLO exposure, respectively, while BA supplementation altered 338 and 440 genes (p < 0.05). Enriched pathways included peroxisome proliferator-activated receptor (PPAR) signaling, fatty acid metabolism, pyruvate metabolism, glycolysis/gluconeogenesis, pentose phosphate pathway, glutathione metabolism, cell adhesion molecules, and adipocytokine signaling. This study adds new evidence on the protective role of BA against CLO-induced toxicity in goslings by combining physiological, histological, metabolomic, and transcriptomic observations. Overall, BA partially mitigated CLO-induced growth inhibition and liver oxidative injury, potentially through coordinated regulation of redox homeostasis, lipid and energy metabolism, and liver metabolic remodeling.

Antioxidants doi: 10.3390/antiox15091090

Authors: Andrés C. Arana-Linares Arley Camilo Patiño Ana Liliana Giraldo Constain H. Salamanca Andres F. Olea

Polyphenolic antioxidants are incorporated into pharmaceutical, dermopharmaceutical, and cosmetic products because of their capacity to modulate oxidative stress, inflammation, microbial imbalance, skin aging, wound repair, and tumor-related processes. However, formulation is constrained by chemical instability, limited bioavailability, insufficient skin permeation, and degradation during processing or storage. This review integrates the chemical characteristics, natural sources, extraction approaches, antioxidant mechanisms, and evaluation of curcumin and chlorogenic acid, and critically examines their delivery through liposomes, niosomes, ethosomes, and transethosomes. Curcumin is lipophilic and poorly water-soluble, whereas chlorogenic acid is hydrophilic but permeability-limited. Their antioxidant activity is discussed through hydrogen atom transfer, single-electron transfer, interruption of lipid peroxidation, metal chelation, and localization within lipid interfaces, together with chemical, biomimetic, and cellular assessment methods. Vesicular carriers can improve encapsulation, stability, release control, skin interaction, biological performance, and incorporation into semisolid dosage forms. However, these benefits are accompanied by formulation-dependent trade-offs involving manufacturing complexity and cost, long-term stability and reproducibility, excipient-related skin tolerability, scale-up, and an application scope that depends on the intended dermal-delivery endpoint. Therefore, efficacy depends on the interplay among antioxidant properties, vesicle architecture, excipient selection, and processing conditions. Curcumin-loaded vesicles are better documented than chlorogenic-acid-loaded systems, particularly for deformable carriers. Future progress requires quality-by-design strategies, standardized characterization, predictive skin models, long-term stability and safety studies, and scalable manufacturing. Overall, antioxidant-loaded vesicles represent multifunctional platforms for developing stable and effective pharmaceutical and cosmetic products.

Antioxidants doi: 10.3390/antiox15091089

Authors: Cristian Paz Muhammad Javid Iqbal Andrea Cristina Paula Lima Alejandro Luarte Pablo Lazcano Ursula Wyneken María Isabel Behrens Daniela Ponce Nicole Jeraldo Sigisfredo Garnica Cecilia Villegas Vaderament-A. Nchiozem-Ngnitedem Bernd Schmidt Eric Sperlich Nicole Cortez Viviana Burgos

Neurodegenerative diseases remain a major therapeutic challenge, with oxidative stress playing a central role in central and peripheral immune system dysfunction that leads to neuronal loss. Natural products from extreme environments represent an underexplored source of neuroprotective agents. In this study, viridicatin, a quinoline-derived alkaloid, was isolated from the Antarctic fungus Penicillium sp. collected from sediments taken from Deception Island, and its structure was unambiguously confirmed by 1D/2D-NMR spectroscopy and single-crystal X-ray diffraction. Viridicatin (100 µM) significantly attenuated H2O2-induced cytotoxicity in HMC-3 human microglial cells, preserving cell viability and mitochondrial membrane potential. Viridicatin modulated the Nrf2 antioxidant signaling pathway, accompanied by increased expression of the downstream antioxidant enzymes HO-1 and NQO1. Moreover, molecular docking revealed preferential binding to the KEAP1 Kelch domain (−8.0 kcal/mol), suggesting indirect Nrf2 pathway modulation. A 100 ns molecular dynamics simulation with MM-GBSA analysis supported the stability of the viridicatin–KEAP1 complex. Notably, viridicatin rescued peripheral immune cells, i.e., peripheral blood mononuclear cells (PBMCs) obtained from older adults with mild cognitive impairment from H2O2-induced cell death, bridging the gap between in vitro mechanistic evidence and clinically relevant human cellular models. This is the first report of neuroprotective activity for viridicatin, positioning this Antarctic-derived alkaloid as a compelling candidate for further preclinical development against age-related neurodegeneration.

Antioxidants doi: 10.3390/antiox15091088

Authors: Zishu Luo Jianglong Zhou Yijia Zhang Huan Li Rong Zhou Ting Zhou Yanxin Zhang Jun You Linhai Wang

Sesame is a significant oilseed crop whose exceptional oxidative stability is closely associated with its abundant endogenous antioxidants. As one of the most predominant lipid-soluble antioxidants in sesame, vitamin E (VE) plays a critical role in scavenging lipid peroxyl radicals, terminating lipid peroxidation chain reactions, and protecting cellular membranes from oxidative damage, thereby maintaining intracellular redox homeostasis and attenuating the development of oxidative stress-related disorders. Although VE is a potent natural antioxidant with significant pharmacological activities in mitigating oxidative stress-related disorders, the genetic mechanisms underlying the natural variation in VE content in sesame remain incompletely understood. Here, variation in VE content was evaluated across 400 sesame accessions grown in two environments. Ultra-high-performance liquid chromatography (UHPLC) analysis revealed that only γ-tocopherol was detected in sesame seeds, with concentrations between 169.33 and 463.31 mg/kg, averaging 316.28 mg/kg. The newly acquired SNP and InDel data from whole-genome resequencing were associated with the phenotypic data, leading to the identification of five significant loci associated with VE content. Comparative transcriptomic profiling of two sesame accessions with distinct VE contents revealed the differential expression of key biosynthetic enzyme genes (such as GGDR, PDS1, VTE2, and VTE4) between the two accessions. By integrating a genome-wide association study with transcriptomic data, two primary candidate effector genes, SINPZ1100015/SiNST1 and SINPZ0901927, were identified, with SiNST1 being particularly prominent. Functional validation further showed that overexpression of SiNST1 in the hairy root of sesame significantly decreased VE content. This investigation advances the comprehension of variations in VE content and the regulatory mechanisms governing its biosynthetic metabolism in sesame, supporting the development of functional sesame varieties for dietary antioxidant supplementation.

Antioxidants doi: 10.3390/antiox15091087

Authors: Flávia G. D. Araújo Henrique L. L. de Araújo Maria da Gloria da Silva João B. Nunes João G. T. L. Resende Fernanda L. Silva Miguel G. Cardoso Eder A. Barbosa Andreanne G. Vasconcelos Guilherme D. Brand Tatiana K. S. Borges Amilcar S. Damazo Daniel C. Moreira José R. S. A. Leite

Amphibians of the family Phyllomedusidae produce complex cutaneous secretions with ecological and pharmacological relevance. Here, we investigated the skin morphology of Pithecopus oreades and P. azureus and the antioxidant function of the tryptophyllin PaT-2 in microglial cells. Histological and histochemical analyses revealed a conserved glandular organization, with mucous glands rich in neutral mucopolysaccharides and serous glands containing acidic and sulfated polysaccharides, concentrated predominantly in the dorsal and cephalic regions. MALDI mass spectrometry imaging showed that PaT-2 (m/z 696.4) is heterogeneously distributed. A second ion (m/z 1626.8), assigned to a truncated form of a phylloseptin-like antimicrobial peptide, was predominantly localized in limb tissues and partially overlapped with PaT-2, suggesting co-deployment of antioxidant and antimicrobial molecules on the skin. In BV2 microglial cells, menadione (10 µM) induced oxidative stress reflected by increased oxidized glutathione (GSSG) and higher total glutathione levels, consistent with a compensatory increase in glutathione synthesis. Synthetic PaT-2 (50 µM) was associated with numerically lower GSSG accumulation and a smaller increase in total glutathione when co-administered with menadione, showing no cytotoxicity while demonstrating modulation of endogenous glutathione status. These findings advance the integumentary biology of Pithecopus, suggest differences in peptide distribution between the specimens examined, and extend the antioxidant characterization of PaT-2 to the level of glutathione status, supporting its further investigation as a candidate molecule for oxidative stress-related research.

Antioxidants doi: 10.3390/antiox15091085

Authors: Edina Pandur Zsombor Bence Tóth Levente Tyukodi Ilona Gróf Szilvia Veszelka Mária A. Deli Zsuzsanna Rozmer Imre Huber

Curcumin and natural curcuminoids possess numerous beneficial properties, including antioxidant, antitumor, and anti-inflammatory effects. However, their application is challenging because of instability and rapid metabolism. Cyclic C5-curcuminoids, a subgroup of curcuminoids, are chemically more stable and less susceptible to hydrolysis or metabolism. In our study, seven synthetic compounds—five cyclic C5-curcuminoids (compounds 8, 9, 11, 12, and 13) and two cyclic chalcones (compounds 4 and 5)—were examined using an in vitro 6-hydroxydopamine-induced neuronal damage model established with all-trans retinoic acid-differentiated SH-SY5Y and BV-2 microglial cells. The antioxidant, anti-apoptotic (including ferroptosis markers (iron, GSH/GSSG, and malondialdehyde)), apoptotic (PARP, cytochrome c, caspase 9, active caspase-3, and oligonucleosome release), and anti-inflammatory effects of these compounds were also investigated. Compounds 5, 9, 12, and 13 significantly decreased reactive oxygen species production (percentage changes: 5: 28.9%; 9: 32%; 12: 29.8%; 13: 49.2%) and increased antioxidant capacity (percentage changes: 5: 63.4%; 9: 85.8%; 12: 31.2%; 13: 85.9%) and antioxidant enzyme activity (catalase, superoxide dismutase, and glutathione peroxidase) in SH-SY5Y cells. Compounds 5, 9, and 12 decreased microglial activation by downregulating Iba1 expression (fold changes: 5: 0.73; 9: 0.39; 12: 0.33) and decreasing glutamate concentration (percentage changes: 5: 26.5%; 9: 41.2%; 12: 27.3%). Together with compound 13, they reduced pro-inflammatory cytokine (IL-6, TNFα) secretion and induced anti-inflammatory cytokine (IL-10) release in SH-SY5Y and BV-2 cells. Based on these results, the blood–brain barrier penetrations of compounds 5, 9, 12, and 13 were also investigated. The highest blood–brain barrier penetration was observed for compound 12 (Papp value 13.6 × 10−6 cm/s). Synthetic cyclic C5-curcuminoids derived from curcumin overcome the limitations of stability and bioavailability and exhibit strong antioxidant, anti-inflammatory, and anti-apoptotic effects, making them promising candidates for treating neurodegenerative diseases.

Antioxidants doi: 10.3390/antiox15091086

Authors: Qingzhe Meng Yongjin Liu Xiaohong Duan Xifei Zhang Guijiang Wang Lingjiang Min Eslam M. Bastawy Fei Luo Zhendong Zhu

Oxidative damage is closely associated with the gradual decline in boar sperm quality during liquid storage at 17 °C. To determine whether L-histidine (L-His) could limit this deterioration, semen was diluted with an extender containing 0, 1, 10, 100, or 1000 μM L-His. Motility and acrosomal integrity were measured throughout storage. On day 7, mitochondrial membrane potential (MMP) and ATP content were determined together with intracellular probe oxidation, membrane lipid oxidation, and Annexin V-FITC/PI staining patterns. The stored sperm were subsequently exposed to capacitating conditions and tested for their ability to bind to oviductal explants. The most favorable responses were observed with 100 μM L-His. Compared with untreated semen, this group retained higher motility and acrosomal integrity and showed higher MMP and ATP content (p < 0.05). It also exhibited less intracellular probe oxidation and membrane lipid oxidation, together with lower proportions of Annexin V-positive and membrane-compromised sperm. Following capacitation induction, sperm stored with 100 μM L-His showed increased tyrosine phosphorylation and higher proportions of capacitated sperm. Their binding index to oviductal explants was also higher than that of the control group. These findings indicate that supplementation with 100 μM L-His can improve the preservation quality of boar sperm during extended liquid storage at 17 °C and maintain functional characteristics of stored sperm, providing new insights into the potential use of L-His as a semen extender supplement for prolonged storage.

Antioxidants doi: 10.3390/antiox15091084

Authors: Mohammad S. Mohammad Sally S. Sakr Marwa A. Kamel Amira A. Gamal Eman S. Abou-Amra Asmahan A. Ali Marwa M. El-Said

Medemia argun is a rare desert palm rich in phenolic compounds with promising biological activities; however, the instability and poor compatibility of phenols in dairy systems limit their functional application. This study characterized the phenolic composition and biological activities of M. argun phenolic extract (MAPE), developed sodium alginate–whey protein concentrate (SAlg/WP) hydrogel beads for phenolic encapsulation, and evaluated their application in fortified yoghurt. HPLC identified quercetin and gallic acid as the predominant phenolic compounds. Antioxidant, antimicrobial, antiviral, and molecular docking analyses were performed, while SAlg/WP–MAPE beads were characterized by encapsulation efficiency analysis, FTIR, and scanning electron microscopy. Fortified yoghurt containing different concentrations of encapsulated MAPE was evaluated for color, syneresis, pH, and antioxidant activity during 14 days of refrigerated storage. MAPE exhibited strong antioxidant and broad-spectrum antimicrobial activities against bacterial and fungal strains, as well as promising antiviral activity against hepatitis A virus (HAV), with an IC50 of 104.89 μg/mL and a selectivity index of 1.6. Molecular docking revealed favorable interactions of rutin, chlorogenic acid, and rosmarinic acid with the HAV 3C protease. SAlg/WP beads achieved 78.92% encapsulation efficiency, while FTIR and SEM confirmed successful phenolic entrapment. Encapsulation effectively preserved MAPE functionality and significantly enhanced yoghurt antioxidant activity during storage, demonstrating its potential for functional dairy applications.

Antioxidants doi: 10.3390/antiox15091081

Authors: Jianchi Lun Yuxin Yan Junji Huang Rong Chen Yimu Ma Mengjie Liu Qian Qu Weijie Lv Shining Guo

Escherichia coli-induced enteritis imposes a substantial economic burden on the global livestock industry. In the current context of reducing and restricting antibiotic use, there is an urgent need for novel therapeutic strategies. Based on traditional Chinese medicine compatibility, anemoside B4 (AB4) and oleanolic acid (OA) form nanoscale assemblies (AB4-OA NPs) through hydrogen bonds and van der Waals forces. Physicochemical characterization quantified the NPs as spherical particles with an average hydrodynamic diameter of 164.16 nm, a PDI of 0.280, zeta potential of −22.16 mV, and stable particle size for 28 days; the hemolysis ratio remained below 3.38% at concentrations up to 2 mg/mL, confirming favorable biosafety. In vitro assays confirmed that AB4-OA NPs outperformed AB4/OA physical mixtures in anti-inflammatory and antioxidant activity by restraining pro-inflammatory factors and activating antioxidant enzymes, verifying nanotechnology’s potency in boosting their bioactivity. In mice challenged intraperitoneally with E. coli, AB4-OA NPs elevated SOD, CAT, GSH-Px and lowered MDA; they inhibited the NF-κB pathway, activated the Nrf2 pathway, balanced inflammatory cytokines, alleviated intestinal leakage, upregulated tight junction proteins and reshaped gut microbiota by reducing Proteobacteria and enriching Firmicutes. Spearman’s correlation study indicated a strong positive relationship between Lactobacillus and Faecalibacterium with intestinal barrier function (Occludin and ZO-1) as well as antioxidant capacity, while Enterococcus was significantly and positively correlated with pro-inflammatory cytokines. These findings demonstrate that AB4-OA NPs exert pronounced protective therapeutic effects on E. coli-induced enteritis via inhibition of inflammation, attenuation of oxidative stress, protection of the intestinal barrier, and regulation of gut microbiota, and that these effects were greater than those of the corresponding physical mixture. This study proposes a novel therapeutic strategy for managing E. coli-induced enteritis and highlights the therapeutic potential of phytochemical-based self-assembled nanomedicines.

Antioxidants doi: 10.3390/antiox15091083

Authors: Thitinun Tarathipayakul Pattaranee Srichairatanakool Pornpawee Sreechomphu Vanichaya Sinpiang Onsaya Kerdto Peraphan Pothacharoen Somdet Srichairatanakool Wachiraporn Tipsuwan

Gardenia sootepensis Hutch. (Golden Gardinia) has received limited scientific attention despite the chemical diversity of its specialized metabolites. In this study, aqueous and 70% ethanolic preparations derived from the leaves, peel plus pulp, and seeds were systematically evaluated for extraction efficiency, total phenolic content (TPC), flavonoid content (TFC), ABTS and DPPH radical-scavenging capacity, HPLC–ESI–MS phytochemical profiles, and cytocompatibility with C28/I2 cells and primary human articular chondrocytes. The largest extract recovery was obtained from seed material, whereas the ethanolic leaf preparation contained the greatest TPC (81.08 ± 4.60 mg GAE/g), and TFC (28.36 ± 3.63 mg QE/g), and showed the strongest ABTS antioxidant activity (272.92 ± 10.95 mg TE/g). HPLC–ESI–MS analysis based on nominal-mass measurements enable the provisional annotation of 34 phytochemical features encompassing iridoids and iridoid glycosides, phenolic acids, flavonoids and their glycosides, xanthones, and benzophenone-related constituents. Overall, extraction with aqueous ethanol yielded a wider spectrum of detectable metabolites than extraction with water, while seed-derived preparations displayed the greatest chemical diversity. Under the experimental conditions and concentration range tested, viability of both immortalized C28/I2 cells and primary human articular chondrocytes remained above 80%. Collectively, these findings demonstrate that the botanical tissue selected and the extraction solvent substantially influence the phytochemical composition and antioxidant properties of G. sootepensis. The observed cytocompatibility provides a basis for subsequent mechanistic investigations but does not establish chondroprotective activity. Further confirmation and quantitative characterization of the provisionally annotated metabolites using high-resolution MS/MS and authentic reference standards are warranted.

Antioxidants doi: 10.3390/antiox15091082

Authors: Ziyang Zhang Kehan Wang Hao Chen Yu Shi Huihui Wang

Nucleus pulposus cells (NPCs) age mainly because reactive oxygen species (ROS) build up too much. Excess ROS is associated with intervertebral disc degeneration (IVDD). In this study, we prepared ergothioneine-derived carbon dots (EGT-Fe-CDs) as an antioxidant nanozyme. EGT-Fe-CDs showed good biocompatibility, superoxide dismutase-like and catalase-like activities, and high total antioxidant capacity. In H2O2-treated NPCs, EGT-Fe-CDs reduced cell damage. Local EGT-Fe-CDs treatment also reduced degeneration in the animal model, based on imaging, disc height index (DHI), and Pfirrmann grade. EGT-Fe-CDs also reduced mitochondrial damage associated with excess ROS. They slow down NPC ageing and help control IVDD. Based on these characteristics, the nanozyme could be a useful option for clinical therapy in the future.

Antioxidants doi: 10.3390/antiox15091080

Authors: Sylmara Esther Negrini-Ferrari Weitao Wang Olga Pol

Fibromyalgia is a chronic nociplastic pain disorder characterized by widespread pain and affective disturbances, in which oxidative stress and neuroinflammation are now recognized as central pathogenic mechanisms. However, current therapies provide only limited symptomatic relief. This study compared the therapeutic and molecular effects of dimethyl fumarate (DMF), an activator of nuclear factor erythroid 2-related factor 2 (NRF2), and cobalt protoporphyrin IX (CoPP), an inducer of heme oxygenase-1 (HO-1), in a reserpine-induced mouse model of fibromyalgia-like pain. Male and female C57BL/6J mice received repeated reserpine administration to induce mechanical allodynia, thermal hyperalgesia, cold allodynia, and depressive-like behaviors. Therapeutic efficacy was evaluated by behavioral testing and molecular analyses of the periaqueductal gray (PAG), anterior cingulate cortex (ACC), and spinal cord (SC). DMF produced a faster antinociceptive response than CoPP, although both compounds ultimately reversed nociceptive hypersensitivity and depressive-like behaviors in male and female mice. These behavioral alterations were associated with increased NLRP3 expression and activated AKT signaling, together with region-specific redox dysregulation involving differential regulation of NADPH oxidases and endogenous antioxidant defenses. Both treatments differentially modulated region-specific redox alterations, enhanced selected antioxidant defenses, and attenuated inflammatory and nociception-related signaling. These findings identify region-specific redox dysregulation as a prominent feature of fibromyalgia-like pathology, providing new mechanistic insight into the molecular basis of nociplastic pain. Collectively, these results demonstrate that DMF and CoPP treatments remodel region-specific redox networks while inhibiting both sensory and affective manifestations of fibromyalgia-like pain, supporting endogenous antioxidant pathways as promising mechanism-based therapeutic targets for fibromyalgia.

Antioxidants doi: 10.3390/antiox15091079

Authors: Xin-Ru Lyu Si-Tao Xu Na Su Min Lin Zi-Ya Zhao Zi-Han Xu Xiang Li Zhi-Hui Lu Tong-Tong Wei Shi-Yu Zhang Qiang Fu Guang-Ji Wang Ying Peng Jian-Guo Sun

Acute liver injury (ALI) is a severe clinical syndrome characterized by systemic inflammation and mitochondrial oxidative stress. This study investigated the hepatoprotective effects of paeonol (PA) in LPS/D-GalN-induced ALI and explored the potential involvement of macrophage mitochondrial function and NDUFS7. Proteomic analysis showed that the expression of mitochondrial respiratory chain-related proteins was suppressed in the liver of ALI mice, while PA treatment was associated with increased expression of several of these proteins. Integrated metabolomic analysis further indicated that PA modulated metabolites associated with energy metabolism and redox processes. In vitro experiments suggested that inflammatory RAW264.7 macrophages contributed to hepatocyte injury, whereas PA attenuated inflammatory responses and reduced macrophage-mediated injury to AML12 cells. In RAW264.7 cells, PA attenuated ROS accumulation and modulated multiple mitochondrial functional parameters, including mitochondrial membrane potential, ATP levels, the NAD+/NADH ratio, and complex I activity. Metabolic studies in human liver microsomes and primary human hepatocytes identified three major PA glucuronide metabolite products with five possible structural assignments. Molecular docking and microscale thermophoresis further indicated that several proposed PA metabolite structures exhibited more favorable predicted interactions and stronger binding to recombinant NDUFS7 than the parent PA. Collectively, these findings support the hepatoprotective effects of PA and its modulation of macrophage mitochondrial function, as well as the potential involvement of PA metabolites and NDUFS7, while further functional validation is required to establish their causal roles.

Antioxidants doi: 10.3390/antiox15091078

Authors: Janu Newar Elizabeth Brees Erika T. Lin Abhik Chakraborty Aliyah Abanes Kathleen M. Fisch William Funk Karen K. Mestan

Maternal smoking during pregnancy (MSDP) remains a significant public health concern. The molecular mechanisms underlying smoking-induced maternal-fetal impact remain incompletely understood. Newer omics platforms, such as cord blood adductomics, can provide preliminary insight into the “perinatal exposome” leading up to birth. The objective of this study is to explore the adductomic signatures of oxidant stress in cord blood associated with MSDP. In a cohort of mother-infant dyads enrolled at a single birth center in Chicago, IL (2008–2021), we conducted a secondary analysis of self-reported maternal smoking exposure data that was linked to an existent adductomics database of 105 addition products (adducts) measured in cord blood plasma. Principal component analysis (PCA), volcano plots and PERMANOVA were used to visualize the variability in adductomic profiles and to compare significantly increased and decreased adduct concentrations according to self-reported current-smokers, former-smokers, and never-smokers. N = 158 participants (N = 6 current-smokers, 26 former-smokers, and 126 never-smokers) had documented self-reported smoking status and were included in the analysis. PCA demonstrated separation between current-smokers and the other groups (R2 = 0.026 for current vs. never-smokers, R2 = 0.115 for current vs. former-smokers; Padj = 0.0075 for both) and no difference between former vs. never-smokers. Among 56 annotated adducts, 27 were increased in current vs. never-smokers, and 32 in current vs. former-smokers. Mode of delivery (p = 0.02) and maternal hypertension (p = 0.001) were significantly different among the three groups. Matched analysis by chronic hypertension (yes/no) supported the main finding that current-smoker adductomic profiles were distinct from both former (p = 0.015) and never-smoker (p = 0.0225) profiles. Self-reported MSDP appears to be associated with cord blood adductomic signatures of oxidant stress. Further studies are needed to determine how the duration, extent and timing of exposure correlate with exposomic changes at birth.

Antioxidants doi: 10.3390/antiox15091075

Authors: Fatemeh Taktaz Salar Hafez-Ghoran

DNA methylation clocks provide a tractable molecular readout for testing whether nutritional and natural-product interventions can modify biological aging. Natural-product-derived antioxidants are biologically plausible candidates because they influence redox signaling, inflammation, mitochondrial function, microbial metabolism and epigenetic regulation, yet their effects on DNA methylation-based aging remain difficult to interpret. Here, we systematically synthesize human intervention studies evaluating antioxidant-rich dietary patterns, botanical and food-derived extracts, marine omega-3 fatty acids, multi-component nutraceuticals and related lifestyle-based interventions with DNA methylation-clock outcomes. The available evidence does not support a uniform epigenetic anti-aging effect. Instead, methylation-age responses were clock-specific, exposure-dependent and often most apparent in metabolically or biologically responsive subgroups. Longer randomized or trial-embedded studies provided the most credible signals, whereas small uncontrolled studies mainly generated hypotheses. Future trials should move beyond claims of epigenetic age reversal and test whether objectively verified natural-product-derived antioxidant exposures produce reproducible, mechanistically linked, and clinically meaningful changes in aging biology.

Antioxidants doi: 10.3390/antiox15091077

Authors: Hawa Sidibé Mojgan Morvaridzadeh Tamàs Fülöp Hicham Berrougui Slimane Belbraouet Michel Nguyen Abdelouahed Khalil

Diet and lifestyle are modifiable determinants of oxidative balance through their influence on exposure to antioxidant and pro-oxidant factors. The oxidative balance score (OBS) is a composite index that reflects the balance between these exposures, with higher scores indicating a predominance of antioxidant factors. In this exploratory pilot study, we examined three OBSs (nutrient-, food-, and lifestyle-based), as well as their combined versions (nutrient–lifestyle and food–lifestyle), across clinical subgroups and evaluated their associations with biomarkers of oxidative stress, inflammation, lipid metabolism, and liver-related biomarkers. A total of 44 older adults were enrolled and stratified into three subgroups (16 healthy, 14 hypercholesterolemia, and 14 post–myocardial infarction). Each participant completed a questionnaire, a 3-day food record, and underwent blood sampling. OBSs were calculated based on 15 nutrients, nine food groups, and two lifestyle components. Correlations and multiple linear regression analyses were performed to examine associations between OBSs and the following biomarkers: plasma total antioxidant capacity (TEAC and FRAP), C-reactive protein (CRP), HDL cholesterol, alanine aminotransferase (ALT), and aspartate aminotransferase (AST). The nutrient–lifestyle OBS (OBSN-L) was significantly associated with lower ALT and AST (adjusted ALT β = −1.06; p = 0.048; AST β = −0.54; p = 0.034). A similar association was observed for the nutrient OBS (OBSN) (adjusted AST β = −0.64; p = 0.022). No significant associations were observed for TEAC, FRAP, or CRP. Neither the OBSF nor the OBSF-L was associated with any circulating biomarkers. Higher nutrient-based OBSs, with or without lifestyle integration, were independently associated with lower liver transaminase levels in older adults with varying levels of cardiovascular risk. OBSs may help to capture dietary and lifestyle patterns associated with liver-related biomarkers. These exploratory findings warrant confirmation in larger, prospective studies.

Antioxidants doi: 10.3390/antiox15091076

Authors: Shahabeddin Rezaei Sisi Cao Min Zeng Maria L. Antonius Jillian T. Pierson Ariana H. Bond Allyson Q. McHenry Joshua J. Blakeslee Richard S. Bruno

Dark green leafy vegetables, such as spinach, are rich sources of α-tocopherol (α-T), a lipid-soluble antioxidant, but may contribute little to α-T status due to low intake and limited bioavailability. This study investigated whether egg co-consumption enhances α-T bioavailability from spinach and whether effects differ by egg dose, food matrix, or timing of intake. In a crossover study, healthy adults consumed deuterium-labeled (dx)-spinach (containing 5 mg dx-α-T) alone; or with 1, 2, or 3 hard-boiled eggs; 2 egg whites; or vegetable oil (9.6 g). Two exploratory trials evaluated delayed egg consumption 3 h after spinach intake. Compared with spinach alone, dx-α-T AUC0–72h (µmol/L × h) was 1.5–1.8-times higher when spinach was co-consumed with 1, 2, and 3 eggs (p ≤ 0.05), respectively, with no egg dose-dependent differences (p > 0.05). Co-ingestion of spinach with egg whites or vegetable oil increased dx-α-T AUC0–72h similar to whole eggs (p > 0.05). Delayed egg intake did not affect dx-α-T AUC0–72h relative to spinach alone (p > 0.05). Plant-derived α-T bioavailability can be improved through co-ingestion with other foods but is independent of egg dose and food matrix. Effective absorption requires concurrent intake, indicating that timing and associated digestive processes are key determinants and support food-pairing strategies to improve vitamin E bioavailability and its antioxidant contribution to plant-based diets. Registered at ClinicalTrials.gov (NCT04287816).

Antioxidants doi: 10.3390/antiox15091074

Authors: Yidan Wang Penghao Wei Tinghao Gao Zhiwei Li Caiqin Deng Hanjing Chen Jianliang Zhang Fengcai Zou Haiyang Song

Oxidative stress is a major consequence of environmental, metabolic, and disease-related challenges in poultry and can compromise cellular integrity and function. Although antioxidant defense mechanisms are broadly conserved across vertebrates, oxidative-stress responses in avian cells remain less well characterized than those in mammalian systems. Superoxide dismutase 1 (SOD1) is a key antioxidant enzyme, but how elevated SOD1 expression influences both oxidative injury and stress-responsive transcription in chicken cells remains incompletely understood. In this study, we established a stable SOD1-overexpressing chicken DF-1 fibroblast cell line and induced acute oxidative stress using hydrogen peroxide (H2O2). Cellular responses were evaluated using CCK-8 viability assays and Hoechst 33342/propidium iodide staining, followed by RNA sequencing and transcriptomic analyses. SOD1 overexpression markedly improved cell viability under H2O2 challenge and substantially reduced necrotic cell death, whereas its effect on apoptosis-associated changes was comparatively limited. Transcriptomic profiling showed that H2O2 induced extensive transcriptional reprogramming, while SOD1 overexpression reshaped this response by attenuating a subset of stress-inducible gene programs. Trend clustering further identified an H2O2-responsive module associated mainly with basic amino acid transport and stress-related regulatory processes that was less strongly induced in SOD1-overexpressing cells. Collectively, these findings demonstrate that elevated SOD1 expression enhances resistance to acute oxidative injury and modifies stress-responsive transcription in chicken fibroblasts. This study extends our current understanding of SOD1-mediated antioxidant cytoprotection in an avian cellular model and provides a molecular basis for further investigation of oxidative-stress regulation in poultry.

Antioxidants doi: 10.3390/antiox15091073

Authors: Maria Azam Mingda Liu Beata Jastrzebska

Retinitis pigmentosa (RP) is a genetically heterogeneous group of inherited retinal degenerative disorders characterized by progressive photoreceptor loss and vision impairment, for which broadly applicable mutation-independent therapies remain limited. To examine the therapeutic potential of combined galanin receptor 3 (GALR3) inhibition and antioxidant therapy in a mutation-independent context, we utilized the rd10 mouse model of RP. We first evaluated the effects of individual treatments with the GALR3 antagonist SNAP-37889 and the antioxidant quercetin, followed by a combined treatment regimen to determine whether simultaneous targeting of neuroinflammatory and oxidative stress pathways provides enhanced retinal protection. Treatment efficacy was assessed using functional and morphological analyses, including electroretinography (ERG) to measure retinal function, optical coherence tomography (OCT) to evaluate retinal structure in vivo, and histological and immunohistochemical analyses to quantify photoreceptor survival and markers of retinal oxidative stress and inflammation. Although the expression levels of individual inflammatory and oxidative stress markers did not consistently exhibit additive responses, the combined treatment produced greater photoreceptor survival and preservation of photopic retinal function than either monotherapy alone. These findings support the hypothesis that simultaneous modulation of oxidative stress and neuroinflammation provides greater neuroprotective benefits, establish a foundation for the development of mutation-independent therapeutic strategies for RP, and identify GALR3 as a promising therapeutic target.

Antioxidants doi: 10.3390/antiox15091072

Authors: Anna Guiotto Malak Alghamdi Robyn Hickerson Michael Conneely Hina Choudhary Patricia Brieva Yunsook Lim Alessandra Pecorelli Giuseppe Valacchi

Visible light (VL) accounts for approximately 50% of the solar radiation reaching Earth’s surface and has emerged as a major contributor to skin photoaging and pigmentation disorders, particularly in individuals with darker Fitzpatrick skin phototypes. Increasing evidence suggests that VL exposure induces oxidative stress, inflammation, and melanogenesis by generating reactive oxygen species. In this study, we investigated the protective effects of a topical antioxidant formulation (AOX Mix) containing 15% ascorbic acid, 0.5% ferulic acid, and 1% tocopherol against VL exposure in ex vivo skin biopsies from donors with Fitzpatrick skin phototypes IV–V. To preserve physiological tissue tension and closely replicate in vivo skin responses, human skin explants were maintained using the TenSkin™ culture system. Samples were pretreated with AOX Mix for 30 min, then exposed to VL for 8 h, and collected on Days 2 and 7 for histological and molecular analyses. Our results demonstrated that prolonged exposure to VL disrupted redox homeostasis and induced structural alterations in skin explants, accompanied by increased markers of oxidative stress and pigmentation. In contrast, pre-treatment with AOX Mix significantly attenuated these effects, preserving tissue architecture and reducing molecular indicators of photodamage. In addition, we observed the co-localization of 4-hydroxynonenal and collagen type I, suggesting that oxidative post-translational modification of collagen may contribute to its loss. These findings suggest that antioxidant-based interventions, such as AOX Mix, may be a promising strategy for protecting dark skin against VL exposure.

Antioxidants doi: 10.3390/antiox15091071

Authors: Yao Hu Lihui Li Xingyu Lei Yiji Zhou Wei Gong Mengshan Sun Rong Song

Edible lily bulbs (Lilium lancifolium Thunb.) are highly valued for their rich nutritional profile and significant medicinal properties. To further enhance these beneficial attributes, this study investigates a novel, green post-harvest enhancement strategy using low-dose 60Co-γ irradiation (2–5 Gy) followed by immediate freeze-drying. Experimentally, fresh bulbs were exposed to varying irradiation doses, after which physiological parameters (enzyme activities, phytochemical contents) and metabolic profiles (via LC-MS/MS) were systematically evaluated. The results demonstrated that 4 Gy represented the optimal dose; compared to the non-irradiated control group (0 Gy), the 4 Gy treatment significantly enhanced the antioxidant defense system by eliciting oxidative stress responses, increasing superoxide dismutase (SOD) and catalase (CAT) activities by 1.55- to 1.86-fold, and elevating total phenolics and flavonoids by 1.31- and 2.69-fold, respectively. Untargeted metabolomic analysis identified 42 differential metabolites, revealing that 4 Gy irradiation primarily upregulated the phenylpropanoid and flavonoid biosynthetic pathways. Key antioxidants, specifically rosmarinic acid and the newly detected alkaloid jurubine, exhibited significant accumulation. Integrated correlation analysis and molecular docking simulations elucidated the underlying mechanisms, providing theoretical insights into their transient interactions with DPPH radicals. In conclusion, low-dose 60Co-γ irradiation serves as a sustainable, green post-harvest processing strategy to improve the nutritional quality of edible lily bulbs by activating intrinsic metabolic defense mechanisms.

Antioxidants doi: 10.3390/antiox15091070

Authors: Linjun Zhou Xiajie Chen Yiran Hou Chengfeng Zhang Jian Zhu Bing Li Rui Jia

Salinity is an important environmental factor affecting the physiological homeostasis of freshwater fish, yet the underlying mechanisms in grass carp (Ctenopharyngodon idella) gills remain unclear. Therefore, grass carp were exposed to different salinity levels for 60 days, and gill responses were evaluated using histopathological, ion regulatory, antioxidant, transcriptomic, and metabolomic analyses. Histological observations showed that high salinity (8 g/L) caused marked structural damage to the gill lamellae. Specifically, Na+ and Ca2+ concentrations and Na+/K+-ATPase activity significantly decreased, while K+ concentration and Ca2+-ATPase activity increased, revealing disrupted ion homeostasis. Salinity exposure also led to decreased antioxidant enzyme activities. Integrated omics analysis further demonstrated that a total of 2447 differentially expressed genes and 268 differentially expressed metabolites were identified, with significant enrichment in pathways related to biosynthesis of amino acids, arachidonic acid metabolism, glutathione metabolism, PPAR signaling, and calcium signaling. Notably, the PPAR and calcium signaling pathways showed positive enrichment under salinity stress, suggesting their potential involvement in the regulation of lipid metabolism, energy allocation, and cellular stress responses. Our findings indicated amino acid biosynthesis and arachidonic acid metabolism as key pathways involved in the adaptation of grass carp gills to salinity stress. Overall, chronic salinity exposure caused structural alterations, disrupted ion regulation, altered antioxidant status, and marked transcriptomic and metabolomic changes in grass carp gills, offering new insight into salinity adaptation in freshwater fish.

Antioxidants doi: 10.3390/antiox15091069

Authors: Xuxiang Zhang Jiafei Long Zhijia Wang Yuping Zhang Tonghao Yang Yongmei Jiang Faming Wu Xin Zhang Xuqiang Nie Gang Wang Sha Liu

Total flavonoids (TF) from Artemisia argyi, a traditional edible-medicinal Asteraceae herb, were extracted via a food-grade ternary deep eutectic solvent (TDES, Glycerol/Levulinic Acid/Xylitol = 1:1:1 molar ratio, 30% w/w water) under ultrasound assistance. A hybrid response surface methodology–genetic algorithm–artificial neural network (RSM-GA-ANN) model optimized parameters to deliver a maximum TF yield of 107.7 mg/g—1.5–1.8-fold higher than conventional hydroalcoholic extraction. Greenness was quantified by AGREEprep (score = 0.70) and MoGAPI (score = 80/100), confirming the method’s sustainability, with TDES retaining >84% extraction efficiency over three reuse cycles. Molecular dynamics simulations revealed TDES forms a more stable hydrogen bond network with plant cell walls (average H-bond lifetime: 101.279 ps vs. 46.698 ps for 50% ethanol), a finding validated by density functional theory calculations showing TDES establishes 7–9 hydrogen bonds with cellobiose (the cellulose repeating unit), far exceeding ethanol’s 1–2 hydrogen bonds. Purified TF exhibited potent radical-scavenging activity (DPPH IC50: 0.176 mg/mL; ABTS IC50: 0.159 mg/mL) and multipotent enzyme inhibition (α-glucosidase IC50: 55.31 μg/mL; acetylcholinesterase IC50: 0.618 mg/mL; pancreatic lipase IC50: 0.125 mg/mL). TF also suppressed HCT116, A549, and HepG2 proliferation (IC50 ≈ 50 μg/mL) and protected HepG2 cells against H2O2-induced oxidative damage. As an in silico probe, the predominant quantified flavonoid eupatilin (3.64 mg/g) docked to xanthine oxidase (−7.78 kcal/mol vs. allopurinol −6.86), offering a structural hypothesis for XO interaction without attributing mixture bioactivity to a single compound. This TDES-based platform offers a scalable, green route to valorize A. argyi for functional food and nutraceutical applications.

Antioxidants doi: 10.3390/antiox15091068

Authors: Sirui Du Weifeng Li Meixiu Jiang

Inflammatory diseases are caused by overactivation of the immune system and lead to tissue damage; their high incidence and complications seriously threaten human health. Currently, non-steroidal anti-inflammatory drugs (NSAIDs) and immunosuppressants are the mainstay of clinical practice, but their long-term use can cause gastrointestinal damage, drug resistance, and other problems. In recent years, traditional Chinese medicine has become a research hotspot for the treatment of inflammatory diseases, among which Icariin (ICA), the main active ingredient of Epimedium, has demonstrated significant anti-inflammatory potential. In this review, a comprehensive literature search of PubMed and Web of Science was conducted, and original studies on the anti-inflammatory effects and mechanisms of ICA were included. The roles and mechanisms of ICA in inflammatory diseases in different systems are summarized. Specifically, ICA suppresses NF-κB and MAPK signaling, inhibits NLRP3 inflammasome activation, and activates the Nrf2/HO-1 antioxidant axis, thereby reducing the production of pro-inflammatory cytokines such as TNF-α, IL-6, and IL-1β. Overall, the current evidence indicates broad anti-inflammatory effects of ICA across multiple organ systems, particularly in atherosclerosis and myocardial infarction; however, clinical translation is limited by poor bioavailability and the lack of standardized dosage data. Future efforts should focus on pharmacokinetic optimization and well-designed clinical trials to facilitate its application in inflammatory diseases.

Antioxidants doi: 10.3390/antiox15091067

Authors: Olga Gostyukhina Elina Chelebieva Tatyana Kukhareva Maria Podolskaya Vitaliy Parfenov Andrey Borovkov Aleksandra Andreyeva

Despite the recognized antioxidant properties of phycobiliproteins (PBPs) from Limnospira platensis, their potential application in bivalve aquaculture—particularly regarding their modulation of endogenous antioxidant defenses—remains largely unexplored. This study evaluated the effects of an aqueous PBP extract at concentrations of 2, 20, and 80 µg/mL on antioxidant parameters in the Pacific oyster Magallana gigas following 24, 48 and 96 h exposure periods. Activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), levels of lipid peroxidation (TBARS) and protein oxidation, and expression of corresponding genes in gills and hepatopancreas have been evaluated. PBPs elicited significant effects on prooxidant–antioxidant balance in oyster tissues. Low concentrations (2–20 µg/mL) enhanced SOD and CAT activities and reduced protein oxidation, indicating protective effects. The highest concentration (80 µg/mL) suppressed enzyme activities—particularly GPx—while elevating TBARS, suggesting prooxidant effects at excessive doses. Tissue-specific responses were observed, with hepatopancreas more vulnerable to oxidative stress than gills. Notably, GPx transcription was upregulated at 80 µg/mL of PBP extract despite suppressed activity, implying its post-transcriptional regulatory properties. These findings demonstrate that L. platensis PBPs modulate oyster antioxidant status in a biphasic manner, with moderate doses conferring stimulation and high doses inducing oxidative stress. This study provides the first integrative evidence of PBP effects on enzymatic, and transcriptional antioxidant defenses in a commercially important bivalve, underscoring their promise as functional bioactive additives while emphasizing the need for careful dose optimization.

Antioxidants doi: 10.3390/antiox15091066

Authors: Sara Gomes Espírito Santo Tereza Cristina da Silva Mathieu Vinken Bruno Cogliati Luís Fernando Barbisan Guilherme Ribeiro Romualdo

In the original publication [...]

Antioxidants doi: 10.3390/antiox15091065

Authors: Tom Ryu Jaejun Lee Ji Won Han Hyun Yang Keungmo Yang

Metabolic dysfunction-associated steatotic liver disease (MASLD) requires practical and inexpensive tools for risk stratification. We investigated whether the red cell distribution width (RDW), a routinely available hematologic parameter, predicts adverse clinical outcomes, using data from 352,623 UK Biobank participants, including 165,700 individuals with MASLD. RDW was analyzed as both a continuous and categorical variable using Cox proportional hazards models, restricted cubic splines, inverse probability treatment weighting, landmark, competing risk, and mediation analyses. Over a median follow-up of 13.6 years, each 1% increase in RDW was independently associated with higher risks of all-cause mortality (hazard ratio [HR] 1.14), cardiovascular disease (HR 1.07), cerebrovascular disease (HR 1.07), and chronic kidney disease (HR 1.09) (all p < 0.001). These associations remained consistent across multiple sensitivity analyses, and conventional biomarkers explained a modest proportion of the observed associations. Oxidative stress, together with other systemic processes affecting erythrocyte homeostasis, represents one potential mechanism underlying these associations. Higher RDW was independently associated with adverse clinical outcomes after adjustment for routine liver and metabolic biomarkers, supporting its potential value as a readily available prognostic marker in MASLD.