Abstract
Metabolic dysfunction-associated steatohepatitis (MASH) is a progressive liver disease characterized by lipid metabolism dysregulation, insulin resistance, inflammation, and potential progression to severe liver conditions. Xiaochaihutang (XCHT), a traditional Chinese medicine, has been explored for its therapeutic potential. This study aimed to systematically investigate the role and mechanisms of XCHT in the treatment of MASH. Network pharmacology and ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) were used to identify active components and pathways. In vitro models using free fatty acids (FFAs) and an in vivo methionine-choline-deficient (MCD) diet-induced MASH mouse model were employed to evaluate hepatoprotective effects, metabolic improvements, and gut microbiota modulation. XCHT reduced FFA-induced injury, decreased aspartate aminotransferase (AST)/alanine aminotransferase (ALT) levels, inhibited lipid accumulation (72% reduction in triglyceride, TG), and alleviated oxidative stress (67% decrease in oxygen species, ROS; 92% restoration of glutathione, GSH). It also modulated gut microbiota, increasing beneficial bacteria, and activated the AMP-activated protein kinase (AMPK)/acetyl-CoA carboxylase (ACC) pathway (2.2-fold increase in phosphorylated AMPK, p-AMPK). Quercetin, kaempferol, β-sitosterol, and baicalin were predicted as key active components. XCHT ameliorates MASH through multi-target regulation of the AMPK pathway and gut–liver axis, supporting its potential as a complementary treatment for metabolic liver diseases.