Ginsenoside Rg1 alleviates MASH by targeting GLS2 to enhance PINK1/Parkin-mediated mitophagy and improve mitochondrial function.

Metabolic dysfunction-associated steatohepatitis (MASH) is characterized by profound metabolic dysregulation and hepatic inflammation and represents a major global health burden with limited effective therapeutic options. Increasing evidence suggests that improving mitochondrial function and enhancing mitophagy may offer promising strategies for MASH treatment. Ginsenoside Rg1 (G-Rg1) has been reported to exert potent anti-inflammatory and antioxidant effects; however, its precise molecular mechanisms and targets in MASH remain unclear. In this study, we investigated whether G-Rg1 ameliorates diet-induced MASH by promoting mitophagy and sought to identify its direct molecular target. Mice treated with G-Rg1 were evaluated using histological, biochemical, and indirect calorimetric analyses to assess hepatic steatosis, fibrosis, inflammation, and energy metabolism. Transcriptomic profiling and transmission electron microscopy revealed enhanced mitophagy and improved mitochondrial ultrastructure following G-Rg1 treatment. Virtual screening and molecular docking identified glutaminase 2 (GLS2) as a potential target of G-Rg1, which was subsequently confirmed by drug affinity-responsive target stability, cellular thermal shift, and binding assays. Mechanistically, G-Rg1 activated the GLS2/PINK1/Parkin pathway, leading to increased mitophagy, reduced hepatocellular lipid accumulation, restoration of mitochondrial function, and attenuation of oxidative stress. Notably, GLS2 overexpression recapitulated the protective effects of G-Rg1 both in vitro and in vivo. Collectively, these findings demonstrate that G-Rg1 alleviates MASH by targeting GLS2 to activate PINK1/Parkin-mediated mitophagy, highlighting GLS2-regulated mitophagy as a potential therapeutic target for MASH.