Sex-specific effects of resveratrol and metformin on mitochondrial function and inflammatory activation in a TNF-α-driven human iPSC-derived endothelial dysfunction model

Abstract

Endothelial dysfunction and mitochondrial impairment are key early events in atherogenesis and exhibit sex-specific patterns. Resveratrol and metformin are cardiometabolic modulators with antioxidative and mitochondrial effects, but their sex-dependent actions in human iPSC-derived endothelial cells (iPSC-EC) remain unclear. iPSC-EC generated from peripheral blood mononuclear cells of six male and six female donors were stimulated with TNF-α to induce a pro-atherogenic phenotype and subsequently treated with resveratrol or metformin. Mitochondrial ROS production, mitochondrial content, endothelial permeability, mitochondrial respiration, inflammatory mediators and LDH release were quantified in a sex-stratified manner. In this TNF-α-driven human iPSC-EC inflammatory dysfunction model, male iPSC-EC displayed stronger mitochondrial superoxide signal, inflammatory activation and endothelial barrier disruption, whereas female iPSC-EC showed a trend towards impaired mitochondrial respiration and reduced IL-10. Resveratrol and metformin trend towards reduced selected TNF-α associated endothelial injury markers. Resveratrol and metformin attenuated TNF-α–associated oxidative stress, cytokine secretion, permeability and LDH release, with male cells mainly benefiting from decreased mitochondrial superoxide signal and inflammation, and female cells showing non-significant tendencies towards improved mitochondrial respiration and restoration of IL-10. These findings highlight cellular sex as a critical determinant of endothelial immunometabolic responses to inflammatory stress and suggest that resveratrol- and metformin-based strategies for vascular protection should be evaluated in a sex-aware manner.