Unraveling the potential mechanisms of Xiaochaihu decoction alleviates metabolic associated fatty liver disease (MAFLD) by integrated transcriptomics, metabolomics, and network pharmacology

BACKGROUND: Metabolic associated fatty liver disease (MAFLD) has currently emerged as the predominant form of chronic liver diseases nowadays, with a high morbidity. Nevertheless, the general remission rate of current treatments for MAFLD remains unsatisfactory. The traditional Chinese medicine formulation Xiaochaihu decoction (XCHD) alleviates triglyceride (TG) and total cholesterol (TC) in patients with fatty liver disease, but the precise mechanisms remain to be determined. PURPOSE: This study aimed to investigate the effects and underlying mechanisms of XCHD on MAFLD. METHODS: The components of XCHD and XCHD-containing serum were characterized by UPLC/MS. High-fat diet and glucose-fructose water-induced MAFLD model in mice was established to evaluate the effects of XCHD. Non-targeted metabolomics, RNA-sequencing, and Network pharmacology were performed and integrated in the mice liver. Molecular biology experiments, like Western blot, were taken to investigate its potential mechanisms. Finally, the effects of PPARγ, perilipin 2 (PLIN2), and perilipin 3 (PLIN3) were detected by over-expressed PPARγ or suppressed PPARγ. RESULTS: XCHD markedly alleviates MAFLD by reducing lipid droplets, TC, and TG accumulation in MAFLD mice and free fatty acid induced HepG2 and AML12 cells. Non-targeted metabolomics suggested that XCHD reduced hepatic lysophospholipid, and RNA-sequencing revealed that the underlying mechanism would be related to lipid droplet formation. Molecular biology experiments indicated that XCHD decreased PLIN2 and PLIN3 in vivo and in vitro. Network pharmacology analysis revealed that the mechanism of XCHD against MAFLD involves the PPARγ signaling pathway. Molecular biology experiments indicated that XCHD decreased PPARγ in vivo and in vitro. Overexpression of PPARγ indicated that XCHD exert regulatory effects through PPARγ-mediated inhibition of PLIN2 and PLIN3. However, the inhibitory effects of XCHD on PLIN2 and PLIN3 are not solely dependent on this pathway. CONCLUSION: XCHD alleviates MAFLD by decreasing lysophospholipid, regulating liver lipid droplets formation, and targeting PLIN2/PLIN3. PPARγ was one of the cross in for PLIN2/PLIN3 pathway, which provides novel insights for MAFLD therapy.