Cholestasis is a pathological condition characterized by disruptions in bile flow, leading to the accumulation of
bile acids (BAs) in hepatocytes.
Allocholic acid (ACA), a unique fetal BA known for its potent choleretic effects, reappears during liver regeneration and
carcinogenesis. In this research, we investigated the protective effects and underlying mechanisms of ACA against mice with
cholestasis brought on by α-naphthylisothiocyanate (ANIT). To achieve this, we combined network pharmacology, targeted BA metabolomics, and molecular biology approaches. The results demonstrated that ACA treatment effectively reduced levels of serum AST, ALP, and DBIL, and ameliorated the pathological injury caused by
cholestasis. Network pharmacology analysis suggested that ACA primarily regulated BA and
salt transport, along with the signaling pathway associated with bile secretion, to improve
cholestasis. Subsequently, we examined changes in BA metabolism using UPLC-MS/MS. The findings indicated that ACA pretreatment induced alterations in the size, distribution, and composition of the liver BA pool. Specifically, it reduced the excessive accumulation of BAs, especially
cholic acid (CA),
taurocholic acid (TCA), and β-
muricholic acid (β-MCA), facilitating the restoration of BA homeostasis. Furthermore, ACA pretreatment significantly downregulated the expression of hepatic BA synthase
Cyp8b1, while enhancing the expression of hepatic efflux transporter Mrp4, as well as the renal efflux transporters Mdr1 and Mrp2. These changes collectively contributed to improved BA efflux from the liver and enhanced renal elimination of BAs. In conclusion, ACA demonstrated its potential to ameliorate ANIT-induced liver damage by inhibiting BA synthesis and promoting both BA efflux and renal elimination pathways, thus, restoring BA homeostasis.