Adipocyte extracellular vesicles (AdEVs) promote a proinflammatory and profibrotic profile in human renal and endothelial cells in vitro

OBJECTIVES: In obesity, white adipose tissue (WAT) undergoes hypertrophic and hyperplastic changes that are driven by phenotypical changes in preadipocytes and adipocytes. WAT also causes a chronic inflammatory state that modifies gene expression and the secretome, including the shedding of adipose-derived extracellular vesicles (AdEVs) into the circulation, which may influence distant cell types and modulate their phenotypes. AIM: To evaluate the effects of AdEVs on renal and endothelial cells and determine their impact on gene expression related to inflammation, fibrosis, and endothelial function. METHODS: Human SW872 preadipocytes were differentiated into adipocytes and subsequently characterized. AdEVs were isolated by ultracentrifugation and analyzed according to ISEV guidelines. Recipient human renal (HCD) and endothelial (EA.hy926) cells were exposed to AdEVs for 24 hours. Gene expression of adipokines, cytokines (IL-6, IL-1B), fibrosis markers, neutrophil gelatinase-associated lipocalin (NGAL), and eNOS was assessed by RT-qPCR and western blotting. RESULTS: Differentiated SW872 cells displayed typical adipocyte morphology and accumulated abundant lipid droplets. Isolated AdEVs showed a donut-like morphology, characteristic size distribution, and expression of CD9 and TSG101, consistent with canonical EV markers. Both renal and endothelial cells internalized PKH67-labeled AdEVs and exhibited increased IL-6 and IL-1B expression (p < 0.05). Renal cells demonstrated increased NGAL mRNA levels, whereas endothelial cells showed reduced eNOS mRNA expression following AdEV exposure (p < 0.05). CONCLUSION: AdEVs from SW872 upregulated cytokine (e.g., IL-6) and NGAL expression in renal cells, while reducing eNOS expression in endothelial cells. These findings suggest that AdEVs may influence early inflammatory and vascular responses in obesity.