Objective: This study investigates the correlations between obesity‐related plasma markers and epigenetic/inflammatory changes in white adipose tissue (WAT), focusing on adipose‐derived stem cells (ASCs). We hypothesize that obesity modulates histone H3K27 marks, modified by demethylases (lysine‐specific demethylase 6A and 6B [KDM6A/KDM6B]) and acetylases (CREB–binding protein [CREBBP]/histone acetyltransferase EP300), affecting ASC function.Methods: Serum and visceral WAT (omental region) was collected from male patients (n = 16, 30–50 years old) undergoing elective gastric or bariatric surgery. BMI and obesity markers were correlated with changes in ASCs (transcript expression, proliferation, and secretion) using reverse transcriptase‐polymerase chain reaction.Results: ASCs from individuals with higher BMI exhibited slower proliferation, increased inflammatory profile, and reduced adipogenic potential, with lower expression of key adipogenic genes. H3K27 acetylase transcripts were also negatively correlated with adipogenesis regulators. Moreover, C‐C motif chemokine 2 (CCL2) and KDM6A expression was higher in the group with obesity, as were CREBBP and EP300. Finally, leptin levels positively correlated with serum, WAT, and ASC CCL2 expression. In vitro, leptin exposure enhanced CCL2 expression/secretion and increased KDM6A/KDM6B and EP300 transcription.Conclusions: In vitro leptin exposure enhanced CCL2 expression/secretion and increased KDM6A/KDM6B and EP300 transcription, highlighting how obesity‐driven epigenetic mechanisms, including leptin‐mediated pathways, disrupt ASC plasticity and perpetuate adipose tissue dysfunction, offering novel therapeutic targets for metabolic disease intervention.

Background: Chronic low-grade inflammation in obesity is linked to white adipose tissue (WAT) dysfunction. Plasma lipopolysaccharide (LPS) activates Toll-like receptor 4 (TLR4), triggering NF-κB and worsening these disturbances. Previously, we showed that histone H3 lysine 27 (H3K27) epigenetic modifications affect WAT gene expression in high-fat-diet mice, identifying key pathways in adipose-derived stem cells (ASCs). This study explores whether NF-κB influences H3K27 modifiers in human ASCs and evaluates fish oil (FO) as a modulator. Methods: Human visceral WAT ASCs were stimulated with LPS and treated with FO enriched with eicosapentaenoic acid (EPA). Flow cytometry, PCR array, RT-PCR, and Western blot assays were used. Results: LPS increased NF-κB activity, elevating KDM6B demethylase levels and H3K27 acetylation. These epigenetic modifications in LPS-stimulated ASCs were associated with persistent changes in the expression of genes involved in adipogenesis, metabolic regulation, and inflammation, even after LPS removal and cell differentiation. FO mitigated these effects, reducing H3K27 acetylation and promoting methylation. Conclusions: FO demonstrates potential in modulating inflammation-induced epigenetic changes and preserving adipocyte function.

Background: Chronic low-grade inflammation in obesity is linked to white adipose tissue (WAT) dysfunction. Plasma lipopolysaccharide (LPS) activates Toll-like receptor 4 (TLR4), triggering NF-κB and worsening these disturbances. Previously, we showed that H3K27 epigenetic modifications affect WAT gene expression in high-fat diet mice, identifying key pathways in adipose-derived stem cells (ASCs). This study explores whether NF-κB influences H3K27 modifiers in human ASCs and evaluates fish oil (FO) as a modulator. Methods: Human visceral WAT ASCs were stimulated with LPS and treated with FO enriched with eicosapentaenoic acid (EPA). Flow cytometry, PCR array, RT-PCR, and Western blot assays were used. Results: LPS increased NF-κB activity, elevating KDM6B demethylase levels and H3K27 acetylation. These epigenetic modifications in LPS-stimulated ASCs were associated with persistent changes in the expression of genes involved in adipogenesis, metabolic regulation, and inflammation, even after LPS removal and cell differentiation. FO mitigated these effects, reducing H3K27 acetylation and promoting methylation. Conclusions: FO demonstrates potential in modulating inflammation-induced epigenetic changes and preserving adipocyte function.

Pyriproxyfen (PPF) and diflubenzuron (DFB) are widely used pesticides with metabolic toxicity in humans underexplored. White adipose tissue (WAT) is a potential target for endocrine-disrupting chemicals. We investigated the effects of PPF and DFB on human adipose-derived stem cells (hASCs) redox balance, epigenetic regulation, and adipogenic differentiation. Visceral WAT hASCs were exposed to PPF or DFB (0.01-2 mg/L). Cytotoxicity was observed at ≥1.5 mg/L, with 1 mg/L selected for further experiments. 8-day exposure to PPF or DFB reduced catalase and superoxide dismutase activities while increasing glutathione peroxidase. This was accompanied by 74% increase in mRNA expression of H3K27 demethylase KDM6B and elevated secretion of CCL2 in PPF-exposed cells. During adipogenic differentiation, PPF and DFB upregulated early transcription factors and enhanced lipid accumulation. Differentiated adipocytes exhibited higher proportion of saturated fatty acids and increased leptin secretion, while adiponectin levels remained unchanged. In mature primary adipocytes, PPF suppressed the secretion of leptin and adiponectin, and significantly increased basal lipolysis. our results show endocrine and metabolic disruption induced by non-cytotoxic concentrations of PPF and DFB. PPF upregulated the epigenetic modulator KDM6B and promoted dysregulated adipogenic programming in hASCs, favouring lipid accumulation and a pro-inflammatory, metabolically compromised phenotype.