Integrative transcriptomic and metabolomic analyses of puerarin against cholestatic liver fibrosis

Cholestatic liver fibrosis is a common pathological manifestation of various biliary tract diseases. Given the limited efficacy of current pharmacotherapies, there is a clear need to develop novel and effective treatment strategies. This study aims to evaluate the protective effects of puerarin against cholestatic liver fibrosis. The therapeutic efficacy of puerarin was assessed in a bile duct ligation rat model. Collagen deposition was evaluated using picrosirius red staining and Masson’s trichrome staining. Immunohistochemistry for cytokeratin 7 (CK7) and CK19 was performed to assess bile duct proliferation. Hepatic mRNA expression profiles and metabolic profiles were analyzed by transcriptome sequencing and untargeted metabolomics, respectively. In parallel, in vitro experiments were conducted using HepaRG hepatocyte cultures exposed to a bile acid mixture, with cell proliferative capacity assessed via EdU assays. Puerarin treatment ameliorated serum biochemical abnormalities, histological injury, and bile duct proliferation in the cholestatic liver fibrosis rat model. Mechanistically, puerarin modulated extracellular matrix deposition and regulated the PI3K-Akt signaling pathway. Furthermore, metabolomic analysis indicated that puerarin altered the hepatic bile acid profile, promoting the conjugation of hydrophobic bile acids. In vitro studies demonstrated that puerarin attenuated bile acid mixture-induced hepatotoxicity in hepatocytes. The present study suggests that puerarin may alleviate cholestatic liver fibrosis, potentially by reducing bile duct proliferation, modulating extracellular matrix deposition, and promoting the conjugation of toxic bile acids. These findings indicate that puerarin could be a candidate therapeutic agent for cholestatic liver fibrosis. Further studies are required to fully elucidate its net protective effect on bile acid homeostasis.