Supplementary file 1_Jianpi-Huayu-Jiedu formula improves metabolic dysfunction-associated steatohepatitis through ameliorateing hepatic oxidative stress and lipid accumulation in vivo and in vitro.docx

Hepatic oxidative stress and lipid accumulation are two key features of metabolic dysfunction-associated steatohepatitis (MASH). Jianpi-Huayu-Jiedu Formula (JPD) has been shown to ameliorate oxidative stress and lipid accumulation, suggesting its potential as an effective treatment for MASH. The current study aimed to explore the therapeutic effects of JPD on MASH. Mice fed a methionine- and choline-deficient (MCD) diet and HepG2 cells induced by oleic acid and palmitic acid (OA&PA) or lipopolysaccharide (LPS) were used as in vivo and in vitro models, respectively. Network pharmacology, molecular biology, and transcriptomics were employed to investigate the efficacy and mechanisms of JPD. Network pharmacology analysis indicated that the PI3K-AKT signaling pathway is a potential target of JPD in MASH. Molecular docking also revealed the binding of active components to AKT1 or PPARG. In vitro, JPD significantly reduced LPS-induced inflammation and OA&PA-induced lipid accumulation. In vivo, JPD inhibited MCD-induced liver injury, inflammation, oxidative stress, and lipid accumulation. Transcriptomic analysis further suggested that the PI3K/AKT and PPAR signaling pathways are involved. An AKT inhibitor significantly attenuated the protective effect of JPD. JPD ameliorates hepatic oxidative stress and lipid accumulation by activating both the PI3K/AKT and PPAR pathways. Our study provides a novel strategy for treating MASH with traditional Chinese medicine.