Mitochondria-targeted antioxidant AntiOxBEN2 prevents metabolic dysfunction-associated steatotic liver disease (MASLD) by enhancing fatty acid oxidation and mitochondrial bioenergetics.

Metabolic dysfunction-associated steatotic liver disease (MASLD) affects approximately 30 % of the global population. Its progression is commonly linked to excessive hepatic fat accumulation, elevated oxidative stress, and impaired mitochondrial function. Given the central role of mitochondria in cellular energy metabolism and redox balance, mitochondria-targeted bioactive molecules have emerged as a promising strategy for the prevention and treatment of MASLD. To this end, we develop AntiOxBEN2, a mitochondria-targeted compound generated by conjugating the antioxidant moiety of gallic acid with the lipophilic triphenylphosphonium cation. This design enables selective accumulation of AntiOxBEN2 in the mitochondrial matrix, taking advantage of the organelle's negative membrane potential. In multiple in vitro disease models, AntiOxBEN2 has demonstrated remarkable antioxidant properties, effectively mitigating oxidative stress and preserving mitochondrial function. However, effects on cellular and mitochondrial energy metabolism in vivo remain unexplored. In the present study, we tested whether chronic peripheral administration of AntiOxBEN2 (0.5 or 2.5 mg/kg, 3x/week) could prevent MASLD development in male and female C57BL/6 J mice fed with a 30 % high-fat, 30 % high-sucrose (Western Diet, WD) diet for 16 weeks. Our results demonstrate that AntiOxBEN2 treatment significantly reduced hepatic lipid accumulation in both sexes without affecting body weight. This reduction was accompanied by improvements in mitochondrial function, including enhanced fatty acid oxidation (FAO) and increased activities of mitochondrial electron transport chain (ETC) complexes. Moreover, AntiOxBEN2 administration lowered circulating levels of hepatic damage markers (ALT and AST), as well as insulin and leptin. Notably, a clear sexual dimorphism was observed, with female mice displaying a more pronounced improvement in mitochondrial parameters. Collectively, these findings highlight the therapeutic potential of AntiOxBEN2 for the prevention and/or treatment of MASLD.