Wuling San Ameliorates Lipid Accumulation in Nonalcoholic Fatty Liver Disease by Improving Mitochondrial Dynamics via Interaction with the Protein Kinase B

Abstract Objective: This study aimed to investigate the effects of Wuling San (WLS) on nonalcoholic fatty liver disease (NAFLD) progression and to elucidate its underlying mechanisms, with particular emphasis on mitochondrial dynamics and protein kinase B (AKT)-mediated signaling. Materials and Methods: Ultra-performance liquid chromatography tandem mass spectrometry and network pharmacology were employed to identify the major bioactive components of WLS and predict their potential molecular targets. Both in vivo (db/db mice) and in vitro (free fatty acid-induced alpha mouse liver 12 hepatocyte) models were used to evaluate the effects of WLS. A series of biochemical assays, histological evaluations, metabolomic analyses, mitochondrial function assessments, and molecular docking studies were performed to elucidate the mechanisms of action. Results: WLS significantly reduced hepatic lipid accumulation, improved liver function, and attenuated inflammatory responses. Metabolomic profiling revealed substantial modulation of lipid metabolites, whereas mitochondrial assays confirmed restored dynamics and structural integrity. AKT was identified as a key regulatory target, and inhibition by MK2206 abolished the beneficial effects of WLS and ursolic acid. Conclusions: WLS alleviates NAFLD by enhancing mitochondrial dynamic function through AKT activation, suggesting that it may represent a promising therapeutic strategy for metabolic liver disease.