Search Results (452)

This study employed integrated transcriptomics and proteomics analysis to investigate the potential role of methionine (Met) in regulating the proliferation, differentiation, and lipid deposition of yak intramuscular preadipocytes (YIMA). Five Met concentrations (0, 0.05, 0.5, 5, and 50 mM) were used to establish the Met model of YIMA. The results of Bodipy, Oil Red O, EdU staining, and qPCR showed that the appropriate Met (0.05, 0.5, and 5 mM) supplementation significantly promoted the proliferation and adipogenic differentiation of YIMA, whereas excessive Met (50 mM) markedly inhibited these processes. To further evaluate the mechanism of Met on YIMA, cells supplemented with 0 mM (control, CON), 0.5 mM (moderate) and 50 mM (excessive) Met were selected for the transcriptomic and proteomic analyses. The results showed that moderate Met treatment primarily enriched pathways related to extracellular matrix–receptor interaction and the PI3K/AKT signaling pathway, while excessive Met significantly enriched processes involving abnormal methylation, DNA damage, and metabolic stress. Functional validation experiments further confirmed that Met regulates YIMA proliferation and differentiation by upregulating p-Akt protein expression and activating the PI3K/AKT pathway. These findings provide molecular insights that support improving yak meat quality and IMF content through dietary Met supplementation. Full article

Backgrounds/Objectives: Fatty infiltration (FI) of rotator cuff (RC) muscles is a critical prognostic factor after surgical repair. While the Goutallier–Fuchs grading system is widely used, its reproducibility is often debated. This study aimed to validate a previously reported three-dimensional (3D) magnetic resonance imaging (MRI)-based volumetric quantification method by comparing it with histologic findings in a chronic rotator cuff tear (RCT) rabbit model. Methods: Eighteen shoulders from nine rabbits were randomly assigned to three groups (n = 6 each): repair (A), chronic tear (B), and control (C). In groups A and B, a chronic RCT model was established by detaching the supraspinatus tendon, with group A receiving repair after six weeks. At 12 weeks after repair, 7.0T MRI was performed, and volumetric quantification of intra-muscular fat was performed using semi-automated 3D Slicer software. Histologic fat proportion was measured via Oil Red O staining and ImageJ analysis. Results: The muscle weight and MRI-based muscle volume were significantly lower in group B than group C (p < 0.05). The radiologically measured fat proportion was significantly higher in groups A (1.8 ± 0.8) and B (2.8 ± 0.7) compared to group C (0.5 ± 0.4, p < 0.001). Histologic analysis showed a corresponding pattern (3.0 ± 1.2%, 5.2 ± 1.0%, 1.7 ± 1.0% for groups A, B, and C, respectively; p < 0.001). A strong positive correlation was identified between the radiologic and histologic measurements of FI (r = 0.784, p < 0.001). Conclusions: Direct histologic comparison validates the reliability of 3D MRI-based volumetric quantification for evaluating FI of the RC muscle in a chronic RCT rabbit model. This objective approach may address the inherent limitations of the conventional qualitative grading system. Full article

Background: Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD) is the most prevalent chronic liver disorder globally. Mazdutide has shown clinical benefits in weight management and metabolic regulation, indicating its potential as a therapeutic agent for MASLD. This study aimed to investigate the efficacy and mechanism of action of Mazdutide against early-stage MASLD. Methods: A MASLD mouse model was induced by a 12-week high-fat diet, followed by a 4-week treatment with subcutaneous Mazdutide (100, 200, or 400 μg/kg). In vitro, a cellular MASLD model was established by treating hepatocytes with 1 mM free fatty acids for 24 h, followed by co-treatment with Mazdutide (10, 20, or 50 nM) or the endoplasmic reticulum (ER) stress inhibitor 4-phenylbutyric acid (4-PBA). Serum and hepatic lipid profiles, liver injury markers, and pro-inflammatory cytokines were quantified. Liver histopathology was assessed by hematoxylin and eosin and Oil Red O staining. Protein expression related to ER stress, inflammation, and lipid metabolism was analyzed by immunohistochemistry and Western blot. Results: Compared with the MASLD model group, Mazdutide treatment significantly ameliorated systemic and hepatic lipid metabolism disorders, reduced liver injury markers and hepatic steatosis, and mitigated inflammation and oxidative stress in MASLD mice and hepatocytes (p < 0.05). Mechanistically, Mazdutide alleviated ER stress by modulating the protein kinase R-like endoplasmic reticulum kinase (PERK) pathway, suppressed the nuclear Factor kappa B (NF-κB)-mediated inflammatory response, and downregulated the expression of key lipogenic regulators including sterol regulatory element-binding protein 1 (SREBP-1), CCAAT/enhancer-binding protein beta (C/EBPβ), and peroxisome proliferator-activated receptor gamma (PPARγ) in both models (p < 0.05). Conclusions: Our findings demonstrate that Mazdutide alleviates hepatic ER stress in MASLD, suppresses inflammatory responses and improved lipid metabolism, which ultimately attenuates disease progression. Full article

Obesity is associated with numerous pathological processes in the body, including inflammation, oxidative stress, and consequently, mitochondrial dysfunction. In recent years, research in anti-obesity therapy has also focused on the function of adipocytes and the inhibition of adipogenesis. In this study, we investigated the effect of the well-known flavonoid quercetin on mitochondrial function, apoptosis and differentiation of human preadipocytes. The Chub-S7 cell line model was used in the in vitro studies. Mitochondrial function was measured by oxygen consumption rates, intracellular ATP content, mitochondrial membrane potential, apoptosis assay (Annexin-5, caspase-9 activity), and ROS generation. Chub-S7 cell differentiation was assessed by Oil Red O staining. The results showed that the quercetin inhibited differentiation of human Chub-S7 preadipocytes and reduced fat accumulation in lipid droplets. Additionally, quercetin influenced mitochondrial biogenesis and mitochondrial uncoupling by changes in mitochondrial respiratory states and also increased mitochondrial membrane potential. Quercetin decreased routine respiration, R/E and netROUTINE control ratio. Our results demonstrate that quercetin is a dietary component that may modulate mitochondrial bioenergetics and inhibit adipogenesis. If these results were confirmed in in vivo studies, quercetin could be considered a factor used to prevent obesity. Full article

Background and objectives: Since postprandial lipid metabolism has emerged as a risk factor for cardiovascular disease, the quality of dietary fat may have a crucial role in atherogenesis and metabolic inflammation. In this study, we propose that the quality of dietary fats and the metabolic status of individuals modulate postprandial triglyceride-rich lipoprotein (TRL) composition and the response of macrophages to TRL. Methods: Randomized controlled crossover trial in the postprandial phase in 12 normal-weight adults and 12 adults with obesity. Each participant consumed both a Western-style (WB) and a Mediterranean-style (MB) breakfast in separate sessions, containing butter or olive oil as the fat source, respectively. Blood samples were collected at baseline (0 h), 2 h, and 4 h postprandially, and TRL were isolated and used to treat THP-1 macrophages. Results: The intake of the WB led to higher concentrations of inflammatory-related markers, particularly in individuals with obesity, and resulted in a higher content of saturated fatty acids and lower of monounsaturated fatty acids in TRL compared to the MB. Staining TRL-treated macrophages with Oil Red O revealed substantial lipid accumulation, which was more pronounced in cells cultured with 4 h TRL from individuals with obesity. This was also evidenced by upregulation of gene expression of lipoprotein uptake receptors following the consumption of the WB. Conclusions: Consumption of a WB led to a more pro-inflammatory postprandial profile and promoted greater lipid accumulation in macrophages, particularly in individuals with obesity, compared to a MB. These findings highlight the importance of fat quality in meals for cardiovascular risk management, especially in populations with obesity. Full article

Proanthocyanidins (PACs) are a key group of bioactive phytochemicals known to provide health benefits. Most PACs are non-bioavailable polymeric molecules that need to be biotransformed by colonic microbes into simple metabolites to exert their pharmacological effects. In this study, six previously unexamined PAC metabolites from Saccharomyces cerevisiae, 3-aminophenol (3-AMP), 3-aminosalicylic acid, 2,4-dihydroxy-6-methylbenzaldehyde, 4-hydroxyphenylacetamide (4-HPA), 3-phenyllactic acid, and 2,4,6-trihydroxyacetophenone, were tested for their antiadipogenic activity using an insulin-dependent 3T3-L1 preadipocyte differentiation model. Lipid accumulation in differentiating preadipocytes was visualized and measured with the Oil Red O assay. Only 3-AMP and 4-HPA significantly reduced lipid accumulation at a concentration of 25 µM. To understand the cellular mechanisms, protein levels of key regulators of adipogenesis and lipid metabolism were analyzed using Western blotting. 3-AMP and 4-HPA may attenuate lipid accumulation by suppressing de novo lipogenesis, with 3-AMP downregulating the peroxisome proliferator-activated receptor (PPAR)-γ/acetyl-CoA carboxylase (ACC)/fatty acid synthase (FAS) axis and 4-HPA primarily inhibiting ACC/FAS signaling. Molecular docking studies indicated that 3-AMP may downregulate PPAR-γ expression through competitive inhibition of insulin receptors. These preliminary findings suggest that 3-AMP and 4-HPA exhibit potential antiadipogenic effects, highlighting PAC-derived postbiotics as promising nutraceuticals for mitigating obesity risk. Full article

This experiment was conducted to explore the effects of betaine on the DNA methylation level, expression level, and fat synthesis of VNN1 in goose hepatocytes by isolating the primary hepatocytes of goose at the cellular level and constructing a fatty degeneration model of goose hepatocytes. In the study, 23-day-old Taizhou goose embryos were used as the research object, and free fatty acid (PA:OA = 2:1) was used to induce steatosis of goose primary hepatocytes. The experiment was randomly divided into seven groups: control group, steatosis model group, and betaine (2, 10, 25, 50, 100mM) group. After 24 h of cell culture, cell viability, oil red O staining, and lipid metabolism-related indicators in cell supernatant were measured, and cells were collected to determine VNN1, FAS, ACC, SCD, SREBPQ gene expression and VNN1 promoter region DNA methylation level. (1) The addition of 0.5 mM fatty acids successfully determined the degeneration of goose hepatocytes. The levels of TG and LDL were significantly increased (p < 0.05), and the level of HDL was significantly decreased. (2) The addition of 100 mM betaine significantly reduced TG levels, and 10, 25, 50, and 100 mM betaine significantly reduced LDL levels. The addition of betaine had no significant effect on HDL level compared to the FFA group (p > 0.05), although a significant overall model effect was observed. Oil red O staining showed that the area of lipid droplets in cells with 50 mM betaine decreased most significantly. (3) The expression levels of VNN1, FAS, ACC, SCD, and SREBPQ genes in the fat model group were significantly higher compared to the control group (p < 0.05), and the DNA methylation level in the promoter region of the VNN1 gene decreased. (4) The addition of 2, 10, 25, 50, and 100 mM betaine significantly reduced the expression of VNN1. The expression of FAS in 2, 10, and 25 mM betaine groups significantly decreased. Betaine at 10 and 100 mM significantly reduced the expression of SREBPQ, but it showed no significant effect on ACC expression. Addition of 2, 50, and 100 mM betaine led to an increased DNA methylation status at the VNN1 gene promoter region. In summary, the addition of betaine can reduce the expression of fatty acid synthesizing genes such as FAS, SCD, and SREBPQ, down-regulate the expression level of the VNN1 gene, increase the DNA methylation level of the VNN1 promoter region, and reduce lipid deposition in goose liver steatosis cells. Full article

Hyperlipidemia (HLP) is a metabolic dysfunction marked by dysregulated lipid metabolism, which jeopardizes cardiovascular health. The function of autophagy modulated by the AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway in HLP pathogenesis has not been fully elucidated. Thus, this study centered on the impacts of different feeding durations on HLP models. ICR mice were given a high-fat diet (HFD) to induce the model, with durations set at 3, 6, 9, 12, and 15 weeks. Body weight, liver and adipose organ indices, serum and hepatic lipid levels, and pathological changes (assessed by Oil Red O and HE staining) were measured. Related pathway markers were detected via immunofluorescence, quantitative real-time PCR (qPCR), and Western blotting. At week 9, the relative protein expression ratios of P-AMPK/AMPK, P-mTOR/mTOR, and P-ULK1/ULK1 were markedly reduced, while the expression levels of LC3Ⅱ/LC3Ⅰ and P62 proteins were notably elevated, exhibiting transient dysregulation characteristics and suggesting a potential optimal modeling time point. It clarifies the temporal pattern, core molecular mechanism, and critical turning point of abdominal adipose metabolic disorder induced by a high-fat diet (HFD) in ICR mice. This study offers a credible basis for the optimal duration of HLP modeling and in vivo animal experimental design. Full article

Probiotic strains exhibiting bile salt hydrolase (BSH) activity represent a promising therapeutic strategy for ameliorating metabolic disorders via targeting the gut–liver axis. Herein, we characterized a newly isolated human-derived Enterococcus strain and investigated its therapeutic potential. Genomic analysis confirmed its safety profile, while in vitro assays demonstrated potent BSH activity. Subsequently, C57BL/6J mice fed a high-cholesterol diet were orally administered this strain over an eight-week intervention period. Although the treatment did not significantly reduce serum cholesterol levels, a marked reduction in hepatic lipid droplet accumulation was confirmed by H&E and Oil Red O staining. Mechanistically, the strain’s potent BSH activity likely modulates bile acid metabolism within the gut–liver axis, thereby specifically inhibiting hepatic lipid deposition. Comprehensive histopathological examination of major organs revealed no signs of toxicity, affirming its excellent in vivo safety profile. In summary, E. lactis IOBRA9746 constitutes a safe and BSH-active probiotic candidate, whose primary beneficial effect lies in directly alleviating hepatic lipid accumulation independent of systemic cholesterol modulation. These findings highlight its potential as a novel liver-targeted intervention for preventing metabolic diseases induced by suboptimal dietary patterns. Full article

A species of the genus Tetracera has been used as herbal medicine in traditional Indian Tetracera loureiri medicine. Tetracera loureiri, a plant from the Dilleniaceae family is considered one of the most valuable herbs in Thailand and is native to Southeast Asia. However, the anti-obesity effects of Tetracera loureiri extract have not been reported. In this study, we screened the effect of EtOH extract on lipid accumulation in a 3T3-L1 adipocyte model at various concentrations using Oil Red O staining, and the results were visualized under a light microscope. The fractionation of the soluble CH₂Cl₂ and EtOAc fractions from the EtOH extract revealed that both fractions significantly inhibited lipid accumulation in adipocytes at 2.5, 5, and 10 μg/mL, compared to n-hexane, n-BuOH, and aqueous extracts. Bioactivity-guided fractionation of the CH₂Cl₂ and EtOAc extract led to the phytochemical investigation of 10 secondary metabolites (1–10), and the structure of these compounds was identified using various spectroscopic methods. All isolated compounds were evaluated for their ability to inhibit adipogenesis at a concentration of 2.5, 5, and 10 μM compared with positive control (Orlistat 100 μg/mL); in particular, compounds 1–3, 5, and 7–8 showed 57.39 ± 6.98, 19.35 ± 4.53%, 75.81 ± 1.75%, 17.61 ± 1.62%, 19.83 ± 5.27 and 14.66 ± 3.85% reduction in fat accumulation at 10 μMm in 3T3-L1 adipocytes, respectively. The activity of these compounds also inhibited the secretion of adiponectin and leptin in 3T3-L1 adipocytes, suggesting their role in disrupting adipocyte function and metabolic regulation. Therefore, the results herein provide experimental evidence supporting the potential of Tetracera loureiri extracts as an anti-obesity therapeutic agent. Full article

Although large-scale studies and potential pathways of genes on intramuscular fat (IMF) in livestock have been reported, research on circRNAs in yaks—a unique, low-IMF-content animal species that is native to the Qinghai–Tibetan Plateau—is still lacking. Based on previous high-throughput sequencing results on longissimus dorsi with different IMF content, a novel circRNA encoded by the VPS13C gene (designated as circVPS13C) was found to exhibit significant differential expression. Here, we systematically characterized the function and mechanism of circVPS13C on IMF deposition in yaks by adopting a series of experiments. Sequencing, RNase R processing, and nucleoplasmic separation experiments confirmed the circular structure feature of circVPS13C, and it was predominantly distributed in the cytoplasm. Furthermore, these experiments demonstrated that circVPS13C was mainly distributed in the cytoplasm. The circVPS13C/miR-5606-x/ECHDC3 axis was constructed through ceRNA network analysis and validated by dual-luciferase reporter and rescue experiments. Furthermore, the function of these three potential regulators during IMF deposition was investigated through CCK-8, BODIPY, Oil Red O staining, and qRT-PCR analyses, and results showed that both circVPS13C and miR-5606-x promoted the differentiation and inhibited the proliferation of yak intramuscular preadipocytes, while the function of ECHDC3 was the opposite. In conclusion, circVPS13C could act as a competitive endogenous RNA (ceRNA) sponge to sequester miR-5606-x, thereby relieving the inhibitory effect of miR-5606-x on ECHDC3. Full article

Type 2 diabetes mellitus (T2DM) can be traditionally treated by edible and medicinal species rich in flavonoids and triterpenoids known for their metabolic benefits. Cucumis prophetarum L. has shown antioxidant and antidiabetic properties in decoction extracts. Since solvent polarity strongly influences the extraction of secondary metabolites, this study investigated the hydroalcoholic extracts of C. prophetarum L. to explore their chemical composition and insulin-sensitizing potential. Hydroalcoholic extracts from the leaf, stem, and root of C. prophetarum L. were analyzed by UV-Vis spectroscopy, ATR-FTIR, and UHPLC-ESI-QqTOF–MS/MS to profile their secondary metabolites. The insulin-sensitizing potential of each extract was assessed using an in vitro model of palmitic-acid-induced insulin resistance in L6 skeletal muscle cells, followed by Western blot analysis of key insulin-signaling proteins. Flavonoid glycosides such as apigenin-C,O-dihexoside, apigenin-malonylhexoside, and luteolin-C,O-dihexoside were abundant in leaf and stem extracts, while cucurbitacins predominated in the root. MTT assay confirmed that hydroalcoholic stem and root extracts of C. prophetarum L. were non-cytotoxic to L6 myotubes, whereas the leaf extract reduced viability only at higher concentrations. Oil Red O staining revealed a pronounced decrease in lipid accumulation following stem and root extract treatment. Consistently, the stem extract enhanced insulin signaling through the activation of the IRS-1/PI3K/Akt pathway, while the root extract primarily modulated the AMPK–mTOR pathway. Importantly, both extracts promoted GLUT4 translocation to the plasma membrane, highlighting their complementary mechanisms in restoring insulin sensitivity. Hydroalcoholic extracts of C. prophetarum L. alleviate insulin resistance through multiple molecular mechanisms, with bioactivity and composition differing markedly from previously reported in the decoctions, which highlight a promising source of insulin-sensitizing phytochemicals and underscore the importance of solvent selection in maximizing therapeutic potential. Full article

Background/Objectives: The study investigated the potential of mixed fruits and berries (MFB) as a dietary intervention to mitigate cafeteria (CAF) diet-induced gut microbiome dysbiosis and hepatic dysfunction associated with metabolic syndrome and steatohepatitis (MASH) in an adolescent rat model. Methods: Forty-eight adolescent male Sprague-Dawley rats (n = 3 cages per group (two rats per cage)) were divided into eight experimental groups, where NC received the normal AIN-93G basal diet, PC received the CAF diet and normal AIN-93G basal diet, T₁ and T₂ received MFB supplementation (3% and 6% levels) without CAF exposure, P₁ and P₂ received a MFB (3% and 6% levels) supplementation initiated at the onset of CAF feeding, and I₁ and I₂ received MFB supplementation initiated 2 weeks after CAF feeding. After 6 weeks, cecal 16S rRNA, hepatic histopathology, Oil Red O staining, and metabolic dysfunction-associated steatotic liver disease (MASLD)-related biomarkers (liver enzymes, alanine aminotransferase (ALT), and aspartate aminotransferase (AST)) were analyzed. Results: AST: ALT ratio was the highest in the PC group (3.63, p < 0.05) compared to the MFB groups. Oil Red O staining showed lower hepatic lipid accumulation, and histological analysis demonstrated a marked reduction in portal inflammatory cell infiltration in MFB. Alpha diversity (Simpson Index) decreased in PC (Kruskal–Wallis, p = 0.043). CAF increased Lactobacillus johnsonii (+75%, p < 0.05), while reducing L. murinus and L. intestinalis (~90%, p < 0.05). MFB supplementation restored Bifidobacterium Pseudolongum and increased Akkermansia muciniphila levels in the P₂, I₁, and I₂ groups (~20-fold, p < 0.05). Bacteroides dorei was present in all groups except the PC group. These bacteria presented a positive correlation with key SCFAs. Conclusions: The results from this study indicated that MFB supplementation modulated gut microbiota composition and enhanced SCFA production, thereby strengthening intestinal barrier integrity and reducing gut-derived inflammation. Collectively, these effects attenuated hepatic lipid accumulation and inflammation, highlighting the potential of MFB to restore gut–liver axis homeostasis disrupted by CAF-induced dysbiosis in adolescent rats. Full article

Compositional heterogeneity of oil bodies (OB) from nine high-oleic peanut (HOP) cultivars was systematically characterized. The results demonstrated that nine OB samples exhibited variability in R, G, and B values (red, green, and blue color channels), with the B channel values significantly differing among cultivars, while no significant color variation was observed in their overall appearance. Fats and proteins dominated the dry matter composition of OB, consistent with typical plant OB structural profiles. The high-fat OB of cultivars J572-O, J6-O, Z215-O, and H985-O exhibited outstanding efficiency in loading lipophilic bioactive compounds. OBs from J16-O, G37-O, Z215-O, J572-O, Y37-O, and Y65-O had a distinctive fatty acid profile: high-oleic acid and monounsaturated fatty acids (MUFAs), with reduced linoleic acid, palmitoleic acid, and saturated fatty acids (SFAs). All OB samples contained four tocopherol isomers (α-, β-, γ-, δ-), with α-tocopherol (5.07–12.59 mg/100 g) and γ-tocopherol (6.36–14.81 mg/100 g) as the predominant forms. Essential amino acids (EAAs) and hydrophobic amino acids were detected, with leucine, phenylalanine, and valine being highly abundant. TEAA/TAA and TEAA/TNEAA ratios complied with FAO/WHO standards. J16-O stood out with a balanced fatty acid profile, high tocopherols, and quality protein, making it a promising candidate for functional foods. Full article

Brown adipose tissue (BAT) has emerged as a promising therapeutic target for metabolic disorders such as type 2 diabetes and obesity. To advance research on BAT activation and elucidate the mechanisms underlying adipogenesis, it is crucial to develop a reliable in vitro model. This study aimed to optimize the differentiation of adipose-derived stem cells (ADSCs) into beige adipocytes and to validate the protocol using primary human ADSCs obtained from eight donors. Protocol optimization was first performed with commercial ADSCs, testing more than 30 combinations of adipogenic conditions. Differentiation was assessed by microscopy, Oil Red O staining, and uncoupling protein 1 (UCP1) expression via reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blot. Among the key adipogenic factors, rosiglitazone proved more effective than indomethacin. Extending the induction phase from 4 to 8 days and maintaining dexamethasone throughout the culture markedly enhanced differentiation efficiency. Serum concentration above 5% was inhibitory, while optimal conditions were identified as 5 μM rosiglitazone and 20 μg/mL insulin. The optimized protocol successfully induced beige adipogenesis in ADSCs from eight independent donors, though efficiency varied considerably which could be attributed to individual donor variability. These findings provide a robust in vitro model for studying beige fat biology and highlight the relevance of personalized approaches in metabolic research. Full article