Hu Gan Tang Ameliorates Lipid Deposition of Metabolic‐Associated Fatty Liver Disease by Promoting Autophagy Through the AMPK/mTOR Pathway

Metabolic-associated fatty liver disease (MAFLD) is the most common chronic liver disease in the world, which lacks effective therapies. Hu Gan Tang (HGT), a modified formulation of the traditional Chinese prescription Si Jun Zi Tang based on the principle of "strengthening the spleen, resolving dampness, and purging turbidity," has been clinically used for MAFLD, but the underlying mechanism remains unclear. This study investigated whether HGT mitigates hepatic lipid deposition by enhancing autophagy via the AMPK/mTOR signaling pathway in vivo and in vitro. HGT significantly ameliorated hepatic injury, reduced lipid accumulation, and upregulated autophagy, evidenced by increased Beclin-1, Atg5, LAMP1, and LC3B-II/I ratio alongside decreased p62 expression in both MAFLD animal models and free fatty acid (FFA)-induced HepG2 cells. Notably, HGT suppressed p-mTOR while increasing p-AMPK levels. Mechanistically, co-treatment with Compound C (an AMPK inhibitor) or 3-MA (an autophagy inhibitor) abolished HGT-induced p-AMPK elevation, LC3B-II/I upregulation, and lipid reduction. Notably, HGT counteracted MHY1485 (a selective mTOR activator)-induced p-mTOR elevation and lipid accumulation. Collectively, these findings demonstrate that HGT alleviates hepatic lipid deposition by activating autophagy through the AMPK/mTOR pathway, highlighting its potential as a therapeutic strategy for treating MAFLD.