Buyanghuanwu decoction (BYHWD) attenuates hepatic fibrosis in CCl4-induced mice by regulating bile acid metabolism through activating PPARα  期刊论文  

Ethnopharmacological relevance: Buyanghuanwu decoction (BYHWD), a classical Traditional Chinese Medicine (TCM) formula, has been widely used for treating cardiovascular and cerebrovascular diseases in China. Accumulating evidence confirms its efficacy in ameliorating organ fibrosis (e.g., pulmonary, renal and myocardial fibrosis), providing a solid pharmacological basis for exploring its potential therapeutic value in liver fibrosis. Aim of the study: This study aimed to investigate the anti-liver fibrotic effect of BYHWD in mice and elucidate its underlying molecular mechanisms. Materials and methods: Male C57BL/6J mice were intraperitoneally injected with 15% CCl4 for 6 weeks to establish a hepatic fibrosis model, and BYHWD (6.575 g/kg and 26.3 g/kg) or sorafenib were administered by gavage from week 4. Serum biochemical indicators, histopathological staining (H&E, Masson and Sirius red), and hepatic hydroxyproline content were used to assess hepatic injury and fibrosis. Hepatic stellate cell (HSC) activation and TGF-beta/Smad signaling pathway, and hepatic inflammation were detected by qPCR, Western blot, immunohistochemistry, immunofluorescence, and arachidonic acid metabolomics assay. Non-targeted metabolomics, quantitative bile acid (BA) profiling, and tandem mass tag (TMT)-labeled quantitative proteomics were employed to explore metabolic and molecular mechanisms. Furthermore, the liver-distributed components of BYHWD were identified using UHPLC-Q-Orbitrap HRMS, followed by molecular docking analysis and validation experiments. Results: BYHWD significantly attenuated liver injury, as evidenced by reduced the serum levels of ALT and AST, and mitigated hepatic fibrosis, characterized by decreased collagen deposition and reduced Hyp content. Additionally, BYHWD inhibited HSC activation, lowered the TGF-beta/Smad pathway, alleviated hepatic inflammation, and regulated arachidonic acid metabolism. Metabolomic and BA profiling analyses revealed that BYHWD reversed CCl4-induced BA metabolism dysregulation, including primary/secondary and conjugated/ unconjugated BAs. Proteomic analysis showed the PPAR signaling pathway is a key enriched pathway, and BYHWD upregulated PPAR alpha expression and modulated the levels of BA synthesis-related enzymes (CYP7A1 and CYP8B1). A total of 84 liver-distributed components of BYHWD were identified in mouse liver tissues. Moreover, molecular docking and validation experiments showed that Safflospermidine B, Astragaloside III, Kaempferol-3O-beta-sophoroside, Genistein, Formononetin, Naringenin, and Astragaloside I are potential peroxisome proliferator-activated receptor alpha (PPAR alpha) -targeting active components of BYHWD. Conclusions: Our study demonstrates that BYHWD exerts an anti-liver fibrotic effect by activating PPAR alpha to regulate BA homeostasis through its multiple active components in mouse liver fibrosis model. The present