Search Results (8,699)

Alzheimer’s disease (AD) is a prevalent chronic neurodegenerative disorder; the progression of this disease is driven by cellular determinants such as oxidative stress and dysregulated neurotrophic signaling. Lavender essential oil is traditionally used in aromatherapy for neuronal regulation and neuroprotection, suggesting its potential neuroprotective effects for chronic neurodegenerative disorders like AD. However, the key active constituents responsible for its benefits and the specific molecular pharmacological mechanisms remain unclear. In this study, we isolated myrtenol from lavender essential oil under the guidance of activity evaluation. Its neuroprotective effects were evaluated in PC12 cells via neurite outgrowth, anti-Aβ/H₂O₂ cytotoxicity, and antioxidant assays. Targets and pathways were explored using inhibitor experiments, cell thermal shift assay (CETSA), drug affinity responsive target stability (DARTS), and Western blot. Myrtenol significantly induced neurite outgrowth in PC12 cells and effectively mitigated cytotoxicity and oxidative stress damage induced by Aβ_25–35 and H₂O₂. Mechanistic studies revealed that myrtenol’s effects are associated with the modulation of tyrosine kinase receptor A (TrkA) and insulin-like growth factor-1 receptor (IGF-1R), activating phospholipase C (PLC)/protein kinase C (PKC) and phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathways to jointly mediate neuroprotection effects against the pathology of AD. This study demonstrates that myrtenol as a highly active component of lavender essential oil possesses NGF-like neuritogenic activity and neuroprotective effects. It provides a foundation for understanding the cellular mechanisms of myrtenol as a small-molecule lead for further investigation in neurodegeneration-related research. Full article

Background: Triple-negative breast cancer lacks established targeted therapies, and only a subset of patients achieves a pathologic complete response to neoadjuvant chemotherapy. We aimed to integrate bulk cohorts with an exploratory single-cell multi-omic dataset from only five patients to identify tumor and immune-related features associated with chemotherapy response. Methods: Bulk analyses were performed in two public breast cancer cohorts (GSE76275 and GSE25065) to compare triple-negative versus non-triple-negative tumors and to relate pretreatment transcriptional and inferred immune infiltration patterns to neoadjuvant chemotherapy response. Separately, in a hypothesis-generating single-cell cohort of five triple-negative breast cancers (n = 5; four responders, one non-responder), we performed single-cell RNA sequencing, T cell and B cell receptor sequencing, single-cell ATAC sequencing, and glycosylation tag profiling. Results: In bulk data, triple-negative tumors showed a loss of luminal estrogen receptor-associated programs, higher proliferation, and CIBERSORT-estimated enrichment of myeloid-associated immune fractions compared with non-triple-negative tumors. Chemotherapy response was associated with modest transcriptional shifts and inferred immune composition differences in triple-negative tumors and more pronounced epithelial, stromal, and inflamed immune changes in non-triple-negative disease. Single-cell data suggested that responder tumors were enriched for T and natural killer cells, antigen-presenting myeloid cells, expanded and diverse T and B cell clonotypes, and immune-associated glycosylation signals, whereas the non-responder sample was dominated by epithelial and fibroblast compartments with secretory, adhesion, and potential immune evasion programs. Checkpoint-related analyses reflected expression patterns and predicted ligand–receptor communication, nominating TIGIT–NECTIN2 as a candidate axis for further investigation. Conclusions: Integrating public bulk cohorts with exploratory single-cell multi-omics supports a model in which chemotherapy sensitivity in triple-negative breast cancer is linked to inflamed, antigen-presenting microenvironments and adaptable antitumor immunity, whereas resistance is associated with stromal and tumor dominance. These candidate biomarkers and pathways require validation in larger independent cohorts, and clinical translation is premature given the exploratory single-cell cohort. Full article

The design, production, and study of new poly[ADP-ribose] polymerase 1 (PARP-1) inhibitors have emerged as an interesting exploration area, since PARP-1 is an overexpressed enzyme in several carcinomas. In this sense, menadione, or vitamin K3, is well known for its use in correct blood clotting, and for the generation of reactive oxygen species, but it is important to mention that it has been used as an antineoplastic agent against several cell lines. Related to the last commentary, in this work, four novel molecules (2–5) were produced from menadione through a Michael addition protocol, using 1,2-ethanedithiol, cysteamine, benzene-1,4-dithiol, and 4-aminobenzenethiol as nucleophiles, and menadione (1) as substrate, to evaluate them as plausible candidates to inhibit PARP-1. It is convenient to note that after their production and spectroscopic characterization, both docking and theoretical studies for each compound were conducted, using density functional theory (DFT) with the hybrid method B3LYP with the 6-311G(d,p) basis set. As a complement, the reactivity properties determined by DFT calculations were obtained for all compounds; the results revealed that 2 has the best properties to bind with PARP-1, and 3 offered good results. Hence, the target compounds were evaluated in vitro, determining their activity against PARP-1, using olaparib as a reference. Molecules 2 and 3 displayed the free binding energy values −7.97 and −9.35 kcal/mol, respectively, but 2 has the best IC₅₀ value, 13.76 µM. It is important to highlight that 2 and 3 must be considered as potential new inhibitor agents against PARP-1, exhibiting competitive IC₅₀ values with olaparib. Full article

Background/Objectives: Pulmonary fibrosis is a chronic, progressive lung disease marked by scarring and inflammation, leading to impaired respiratory function. This study aimed to investigate the combined therapeutic effects of pirfenidone (PFD), metformin (MET), and bone marrow-derived mesenchymal stem cells (BM-MSCs) on bleomycin (BLM)-induced pulmonary fibrosis in rats. Methods: Forty-eight Western Albino rats were divided into six groups: normal control, BLM-positive control, and four treatment groups receiving PFD, MET, BM-MSCs, and their combination. Treatments were administered for four weeks starting on day 21 post-BLM instillation. Lung tissues were analyzed for oxidative stress markers, inflammatory cytokines, apoptotic markers, and fibrogenic gene expression. Histopathological changes were assessed using hematoxylin and eosin (H&E) and Masson’s trichrome staining. Results: The combination therapy significantly reduced oxidative stress and inflammatory markers while enhancing antioxidant capacity. It decreased pro-apoptotic Bcl-2-associated X protein (BAX) and increased anti-apoptotic B-cell lymphoma 2 (Bcl-2) levels. Additionally, anti-inflammatory interleukin-10 (IL-10) was elevated, while tumor necrosis factor-alpha (TNF-α) and transforming growth factor-beta 1 (TGF-β1) levels were markedly lowered. Gene expression analysis showed a significant downregulation of matrix metalloproteinase-9 (MMP-9) and collagen type 1 alpha 1 (Col1α1). Histologically, the combination treatment group exhibited minimal fibrosis and inflammation, closely resembling normal lung tissue. Conclusions: The combination of PFD, MET, and BM-MSCs offered superior therapeutic efficacy in treating BLM-induced pulmonary fibrosis compared to individual treatments. This multimodal approach effectively targets oxidative stress, inflammation, apoptosis, and fibrosis, suggesting strong potential for future clinical application. Full article

Background and Aims: The rising global incidence of cholangiocarcinoma (CCA) coincides with epidemics of type 2 diabetes (T2D) and chronic hepatitis B virus (HBV) infection. Although both are established independent risk factors, the shared molecular mechanisms by which they contribute to cholangiocarcinogenesis remain poorly understood. We hypothesized that T2D and HBV converge on a state of chronic metabolic inflammation (“metaflammation”) that drives CCA progression through a conserved transcriptomic network. Methods: We performed an integrative bioinformatics analysis of transcriptomic data from public repositories, including samples of CCA (TCGA-CHOL, n = 45; GSE107943, n = 163), T2D-affected liver (GSE23343, n = 20), and HBV-infected liver (GSE58208, n = 102). Acknowledging that the T2D and HBV datasets were derived from whole-liver tissue, whereas CCA originates in the biliary epithelium, we identified differentially expressed genes (DEGs) across conditions and defined a core gene set shared among them. Subsequent analyses included functional enrichment, construction of protein–protein interaction (PPI) networks, survival analysis, and protein validation. Results: We identified a core metaflammation signature comprising 156 genes that were consistently dysregulated across T2D, HBV, and CCA. Pathway analysis revealed significant enrichment in PPAR signaling, cytokine–cytokine receptor interaction, PI3K-Akt, and TNF signaling pathways. Protein–protein interaction (PPI) network analysis identified IL6, TNF, AKT1, STAT3, and PPARG as the top hub genes. These hubs were functionally modularized into clusters associated with inflammatory signaling, metabolic regulation, and cell growth and survival. In the TCGA CCA cohort, high expression of IL6, TNF, AKT1, and STAT3 and low expression of PPARG correlated with advanced tumor stage and poorer overall survival (e.g., IL6: ρ = 0.42, p = 0.01). A metaflammation score derived from these hubs (weighted combination of the five genes) emerged as an independent prognostic factor (HR = 2.8, p < 0.001). Protein-level dysregulation of these hubs was confirmed via immunohistochemistry. Conclusions: This study defines a conserved metaflammation network that links T2D and HBV to CCA, identifying key hub genes and pathways. This signature provides a mechanistic explanation for epidemiological risks, serves as a novel prognostic tool, and offers a rationale for targeting metaflammation in prevention and therapy for high-risk populations. Full article

Background: Hilar cholangiocarcinoma (HC) is a highly aggressive malignancy with a poor prognosis, highlighting the urgent need to elucidate its molecular drivers. This study aimed to systematically identify and functionally validate key genes and pathways driving HC pathogenesis. Methods: RNA sequencing (RNA-seq) was performed on paired primary HC tumors and matched adjacent non-tumorous tissues to identify differentially expressed genes (DEGs). Subsequent bioinformatic analyses, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, and protein–protein interaction (PPI) network construction, were conducted to characterize the functional landscape and identify hub genes. Transwell assays and orthotopic metastatic models were used to investigate the functions of Contactin-1 (CNTN1) in HC invasion in vitro and metastasis in vivo. Results: RNA-seq analysis identified 35 DEGs in HC, mainly involved in cell adhesion, cytoskeletal regulation, and axon development. PPI network analysis identified six hub genes, including CNTN1, NCAM1, PLP1, GPM6B, SLC1A3, and PMP2. Furthermore, we demonstrated that CNTN1, a neuronal membrane glycoprotein, was markedly up-regulated in HC at both mRNA and protein levels, and its elevated expression correlated with poor prognosis. Gain- and loss-of-function studies demonstrated that CNTN1 promotes HC cell invasion in vitro and metastasis in vivo. Mechanistically, CNTN1 exerts its pro-invasive effects by activating the PI3K-AKT signaling pathway and inducing epithelial–mesenchymal transition (EMT). Conclusions: Our integrated analysis identifies CNTN1 as a critical oncogenic driver in HC, promoting metastasis through PI3K-AKT-mediated EMT. These findings nominate CNTN1 as a potential prognostic biomarker and therapeutic target in HC. Full article

Breast cancer remains a leading cause of cancer-related morbidity and mortality worldwide, with therapeutic response being shaped by the unique biology of each breast cancer subtype. Immunotherapy has emerged as a transformative approach in selected disease subtypes, with the most successful results being found in relation to triple negative breast cancer (TNBC). Immune checkpoint inhibitors (ICIs) have transformed the management of many solid tumours. In breast cancer, they have demonstrated clinical benefit in TNBC when combined with chemotherapy, establishing a new standard of care in both early-stage and metastatic settings. However, the majority of breast cancers exhibit intrinsic or acquired resistance to checkpoint blockade, driven by low tumour immunogenicity and an immunosuppressive tumour microenvironment. Recent advances in cellular immunotherapy could represent the next frontier in the therapeutic landscape of breast cancer. Chimeric antigen receptor (CAR) T cell targeting antigens such as HER2, ROR1, MUC1, mesothelin, and B7-H3 are entering early-phase clinical evaluation with results eagerly awaited. Parallel approaches, including tumour-infiltrating lymphocyte (TIL) therapy, T cell receptor (TCR)-engineered T cells, and CAR-natural killer (CAR-NK) platforms, offer alternative mechanisms to overcome antigen presentation barriers and immune evasion. This review summarises current clinical evidence for immunotherapies in breast cancer, highlights emerging cellular strategies, and discusses key challenges including antigen specificity, off-tumour toxicity, and tumour microenvironment-mediated resistance. Future progress will likely depend on rational combination approaches and next-generation engineered immune cell platforms to achieve durable and personalised clinical benefit. Full article

Studies of natural products and their semisynthetic derivatives are a valuable source of therapeutic agents. The aim of this work was to obtain new 30-phosphoramidate derivatives of betulin and determine their biological potential. The synthetic approach utilized the Staudinger reaction (the introduction of a phosphoramidate group), the Steglich reaction (the introduction of an alkynyl group), and the Jones reaction (the introduction of a carboxyl group). The structures of the target compounds were determined using spectroscopic methods (¹H NMR, ¹³C NMR, ³¹P NMR, and HRMS). The new derivatives were tested for antiproliferative activity against MV4-11, A549, MCF-7, PC-3, and HCT116 cancer cells and against normal MCF-10A cells using the MTT and SRB methods. Apoptosis studies were performed for the most active compounds (6B and 7A), potential molecular targets (AutoDock software) were identified, and lipophilicity parameters (RP-TLC method, SwissADME website) were determined. The greatest effect on apoptosis and caspase 3/7 activation was observed for the diester derivative 7A. Compound 7A showed a high lipophilicity parameter in the study group. Full article

Background: Myocardial ischemic injury, encompassing acute myocardial infarction (MI) and ischemia/reperfusion (I/R) injury, remains a major cause of cardiac morbidity and mortality worldwide, and is driven by interconnected molecular and cellular processes, including cardiomyocyte apoptosis, inflammatory activation, mitochondrial dysfunction, oxidative stress, and impaired angiogenesis. Mesenchymal stem cell (MSC)-derived exosomes have emerged as a promising cell-free nanotherapeutic strategy for cardiac repair due to their ability to transfer bioactive molecules that modulate multiple signaling networks involved in myocardial survival and regeneration. This systematic review aimed to synthesize evidence on the mechanistic basis of MSC-derived exosome mediated cardioprotection in myocardial ischemic injury. Methods: A systematic search of Ovid MEDLINE, Scopus, and Web of Science was conducted to identify studies investigating the effects of MSC-derived exosomes on myocardial ischemic injury. Eligible studies included clinical and preclinical models of MI or I/R injury assessing functional, biochemical, and molecular outcomes. Results: Seven preclinical studies published between 2015 and 2025 met the inclusion criteria. Exosome administration consistently improved cardiac function, reduced infarct size, and preserved myocardial architecture. Biochemical analyses revealed decreased cardiac injury markers, alongside suppressed apoptosis, inflammation, and oxidative stress. Mechanistically, MSC-derived exosomes delivered regulatory miRNAs (e.g., miR-19a, miR-125b, miR-205, miR-294) and lncRNAs (HAND2-AS1) that modulated key signaling pathways including PI3K/Akt, JAK2/STAT3, HAND2-AS1/miR-17-5p/Mfn2, and HIF-1α/VEGF. These molecular effects collectively inhibited apoptotic and inflammatory responses, enhanced mitochondrial integrity, and promoted angiogenesis and myocardial repair. Conclusions: MSC-derived exosomes confer robust cardioprotection against myocardial ischemic injury through integrated anti-apoptotic, anti-inflammatory, antioxidant, and pro-angiogenic mechanisms. Their multifaceted bioactivity, low immunogenicity, and potential for targeted delivery highlight their potential as a next-generation nanomedicine for ischemic heart disease. Future studies should emphasize standardized exosome production, mechanistic profiling, and translational validation in large-animal and clinical models. Full article

Background/Objectives: Eupalinolide B (EB), a natural compound derived from Eupatorium lindleyanum DC, has demonstrated multiple pharmacological activities. However, its role in modulating oxidative stress remains incompletely understood. Methods: In this study, we investigated the antioxidant effect and underlying mechanism of EB in lipopolysaccharide (LPS)-induced RAW264.7 macrophages. Results: EB significantly attenuated LPS-induced oxidative stress as evidenced by reduced levels of intracellular reactive oxygen species (ROS), nitric oxide (NO), and malondialdehyde (MDA) alongside enhanced superoxide dismutase (SOD) activity and an increased reduced/oxidized glutathione (GSH/GSSG) ratio. Using activity-based protein profiling, we identified peroxiredoxin 4 (PRDX4) as a key binding target of EB. Direct interaction was confirmed through labeling and competitive binding assays with purified PRDX4 protein. High-resolution mass spectrometry revealed that EB covalently binds to Cys54 and Cys248 residues of PRDX4. Furthermore, EB treatment upregulated PRDX4 protein expression in LPS-stimulated RAW264.7 cells. siRNA-mediated knockdown of PRDX4 significantly blunted the antioxidant effects of EB, confirming the functional relevance of this target. Conclusions: Our findings demonstrate that EB alleviates LPS-induced oxidative stress in macrophages by covalently binding to and stabilizing PRDX4, thereby enhancing cellular antioxidant capacity. This study unveils a novel mechanism whereby a natural product enhances cellular antioxidant capacity by covalently stabilizing a key peroxidase, highlighting the potential of EB as a therapeutic agent and PRDX4 as a promising target for oxidative stress-related diseases. Full article

Bovine leukemia virus (BLV) is an oncogenic deltaretrovirus that infects B cells, and its possible presence in humans has garnered increasing attention. This study included 58 participants: 11 with B-cell precursor acute lymphoblastic leukemia (B-ALL, cases) and 47 healthy individuals (controls). Researchers assessed anti-gp51 antibodies and BLV proviral DNA in bone marrow and blood samples. Seropositivity was observed only in the B-ALL group (18.2%; 2/11), while all controls were seronegative. Quantitative PCR targeting the pol gene detected proviral DNA in 74.1% of samples, with similar detection rates between cases and controls. Although proviral load was higher in controls, this difference did not reach statistical significance. Conventional and nested PCR for other viral genes revealed a differential pattern: amplification of the tax gene was significantly associated with B-ALL, whereas gag and env were not. Bayesian Chow–Liu network analyses identified dependencies among viral genes and suggested that contextual factors, such as fieldwork, may influence the association between molecular positivity and B-ALL. Sequence analyses showed that the detected BLV strains clustered with previously reported bovine and human sequences from Colombia, all within genotype 1. These findings support human exposure to BLV and raise important questions about its persistence and potential connections to hematological diseases in humans. Full article

Background: B-cell malignancies have been effectively treated using chimeric antigen receptor-T (CAR-T) treatment employing traditional αβT cells. However, because of several obstacles, application in acute myeloid leukemia (AML) is still restricted. A safer “off-the-shelf” alternative can be supplied by CAR-γδT cells, which have major histocompatibility complex (MHC)-independent tumor identification capabilities and a decreased risk of graft versus host disease (GvHD). This study aimed to develop FLT3-targeted CAR-γδT cells that co-express cytokines (IL-2 or IL-7) to increase their anti-AML persistence and therapeutic efficacy. Methods: FLT3-CAR-γδT cells, FLT3-IL2-CAR-γδT cells, and FLT3-IL7-CAR-γδT cells were constructed. Their antitumor potency was comprehensively assessed through cytotoxicity assays, cytokine release, and persistence evaluation in vitro (using AML cell lines and primary AML cells) and in vivo (via mouse model). Results: Superior cytotoxicity against AML cell lines (OCI-AML3, MOLM-13, THP-1, and MV4-11) was demonstrated by FLT3-IL2-CAR-γδT cells, which also released higher levels of granzyme B, interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α). FLT3-IL2-CAR-γδT cells exhibited cytotoxicity in some primary AML cells in vitro. During the antigen-repeated stimulation assay, FLT3-IL2-CAR-γδT cells preserved the stem cell-like memory T (T_SCM) cell subsets, sustained cytokine release, and maintained excellent viability. FLT3-IL2-CAR-γδT cells considerably slowed the development of AML in vivo and extended the existence (>68 days) of mice. Conclusions: FLT3-IL2-CAR-γδT cells exhibit potent and durable anti-AML activity, providing a novel strategy for clinical AML immunotherapy. Full article

Background: The therapeutic effect and mechanism of protocatechuic aldehyde (PAL) on vascular dementia (VaD) were studied from a multi-group perspective. Methods: The pharmacological property of PAL was assessed by using both an in vivo two-vessel occlusion (2VO) rat model and an in vitro astrocyte–neuron co-culture system with oxygen–glucose deprivation (OGD) injury. On the basis of neurobehavioral test, Morris’ water maze test and hematoxylin and eosin staining, the pathological transformation of cognitive function and ischemic cerebral tissue was assessed. Key metabolites and targets through the comprehensive analysis of brain tissue and plasma metabolomics and transcriptomics were screened. Western blot and immunofluorescence were measured to assess proteins related to glutamate release, lactate shuttle and glycolysis. Results: PAL markedly improved the cognitive dysfunction of 2VO rats and reduced the nerve function score. The degeneration of neurons in the Hippocampal CA1 region was appreciably reduced. A total of eight common metabolites, including L-glutamate and L-glutamine, have been identified from plasma and brain sources. The pathway enrichment of glutamate metabolism is closely related to multiple energy metabolic pathways related to glycolysis. Combined with transcriptomic analysis and in vivo experiments, it was found that PAL can significantly downregulate the expression of the glutamate-releasing protein vGLUT1 and promote the process of glutamate transformation into glutamine. At the same time, it enhances the expression of lactate production, shuttle and utilization of related proteins GLUT-1, HK2, PFK, LDHA/B and PDH, MCT1/2/4. In the subsequent cell co-culture system, we confirmed that PAL can effectively lower the expression of vGLUT1, reduce the content of glutamate, and promote the lactate shuttle process, thus increasing the content of lactate and ATP and reducing apoptosis. Conclusions: PAL is associated with upregulation of key glycolytic enzymes and MCTs, suggesting a potential enhancement of the lactate shuttle mechanism. This process may involve the regulation of glutamate metabolism and coordinated modulation of energy metabolism pathways such as glycolysis, thereby improving intercellular energy supply and contributing to the therapeutic effects observed in vascular dementia. This study provides a mechanistic basis and preclinical evidence for the clinical development of PAL. Full article

Influenza A virus (IAV) infection constitutes a major public health threat. Severe influenza virus infection can induce intense inflammatory responses and lung injury, leading to serious clinical symptoms or even death. The utility of current anti-influenza drugs is often limited by side effects and the emergence of drug-resistant strains. Based on the critical role of L-type voltage-gated calcium channels (L-VGCCs) in influenza virus replication, this study investigates the antiviral activity and mechanism of verapamil, a classic L-type calcium channel antagonist, against H1N1-UI182 virus. Verapamil, an L-type calcium channel blocker, is widely used in the treatment of cardiovascular diseases and has a well-established safety profile. Through molecular dynamics (MD) simulation and network pharmacology analysis, we predicted the stable binding mode of verapamil to the target protein (PDB id: 6JPA) and its potential multi-target network. In vitro, verapamil exhibited antiviral activity against H1N1-UI182 in MDCK cells, enhancing the survival rate of infected cells and reducing viral nucleoprotein (NP) expression. In a lethal H1N1-UI182 infection mouse model, verapamil treatment markedly improved survival rates, alleviated weight loss and lung pathological damage, exhibiting a dose-dependent protective effect. Lung tissue analysis showed that verapamil effectively reduced the lung index and viral load, suppressed the activation of the Nuclear factor kappa B (NF-κB) signaling pathway, and decreased the expression of key inflammatory factors, thereby mitigating the cytokine storm. A comparison of administration regimens indicated that pre-treatment yielded optimal efficacy, suggesting verapamil acts primarily during the early stage of the viral life cycle. This study systematically elucidates that verapamil exerts antiviral and immunomodulatory effects by regulating the NF-κB pathway. Network pharmacology analysis suggested the potential involvement of multiple targets and pathways, including EGFR, SRC, and phospholipase D signaling, providing hypotheses for future mechanistic investigation. This paper supports a drug repurposing strategy against drug-resistant influenza viruses and highlights its significant potential for clinical translation. Full article

Background/Objectives: This article employs both in vivo and in vitro experiments. Methods: The core targets and key pathways of Paeoniflorin were predicted using a PPI network analysis, GO analysis, and KEGG analysis. Subsequently, molecular docking analysis and molecular simulation dynamics were performed on the core effector. In vitro experiments employed a UVB-irradiated B16F10 cell model. The effects of Paeoniflorin on melanin content and tyrosinase activity were evaluated. Apoptosis and inflammatory cytokine levels were also assessed in vitro. In vivo experiments used a model combining progesterone injection with UV irradiation. Histopathological skin changes and melanin granule distribution were examined using HE staining. Skin melanin content, tyrosinase activity, and expression levels of related proteins were measured. Additionally, ELISA assays measured serum IL-6 and TNF-α inflammatory cytokines in mice. Results: Rese screening identified 69 targets involved in Paeoniflorin’s effects on melanogenesis, including TNF-α, IL-6, TP53, MAPK3, HIF1A and BCL2. Molecular docking and molecular dynamics simulations indicate that Paeoniflorin exhibits strong affinity for tumor necrosis factor-α. In in vitro experiments, Paeoniflorin significantly reduced UVB-induced melanin content and tyrosinase activity in B16F10 cells. It also promoted melanocyte apoptosis and a dose-dependent decrease in IL-6 and TNF-α levels. In vivo, Paeoniflorin significantly reduced epidermal and dermal thickness and inhibited inflammatory infiltration. It decreased melanin granules, melanin content, tyrosinase activity, and IL-6 and TNF-α levels in mouse skin tissue. Conclusions: This research indicates that Paeoniflorin can significantly suppress UVB-induced cellular inflammatory responses by inhibiting the TNF signaling pathway, thereby reducing hyperpigmentation. Full article