Ethanol Extract of Wine-Simmered Rhubarb Attenuates Cholestasis Liver Injury via Bile Acid Metabolism and FXR Signaling Pathway

Objective: To investigate the mechanism of ethanol extract of wine-simmered rhubarb (EEWR) in alpha-naphthyl isothiocyanate (ANIT)-induced rats and chenodeoxycholic acid (CDCA)-induced HepG2 cells. Materials and Methods: The extracted components of EEWR were characterized using UPLC-MS. A rat cholestatic liver injury (CLI) model was constructed using ANIT and intervened with EEWR or ursodeoxycholic acid. Evaluation of the affinity between the active ingredients of EEWR and farnesoid X receptor (FXR) using molecular docking and dynamic simulation. Results: The treatment with EEWR demonstrated a notable ability to alleviate ANIT-induced CLI in rats. It led to a decrease in liver function-related biochemical markers, while also ameliorating liver tissue damage and lowering pro-inflammatory cytokine levels. Metabolomic analysis revealed that EEWR influences bile acid synthesis, secretion, and various metabolic pathways associated with bile acids, in addition to impacting other biological functions. In addition, the results of quantitative real-time polymerase chain reaction (qRT-PCR), immunofluorescence, and the results of Western blotting showed that EEWR participated in regulating the synthesis, transport, and metabolism of bile acids by regulating the expression levels of FXR and its downstream factors in vivo and in vitro . The main active ingredients of EEWR are anthraquinones, including chrysophanol, emodin, rhein, and aloe-emodin, all of which have good binding ability to FXR. Molecular dynamics simulation results show that chrysophanol binds to the FXR protein in a stable conformation. Conclusions: The findings of this research indicated that EEWR may mitigate cholestatic liver damage caused by ANIT by modulating the expression of proteins associated with the FXR signaling pathway, diminishing inflammation, and influencing bile acid metabolism.