Abstract: Objective This experiment aimed to investigate the protective effect of Qingchang Ligan Formula(QCLGF), a herbal medicine, against α-naphthyl isothiocyanate(ANIT)-induced cholestatic liver injury(CLI)in mice and its possible mechanism. Methods Twelve mice were randomly divided into control, model and QCLGF intervention group, with 4 mice in each group. A mouse model of CLI was established by intragastric administration of ANIT, and mice in the intervention group were treated with QCLGF. Western blotting was conducted to detect expression of autophagy-related proteins in mouse liver tissues, including coiled-coil myosin-like BCL2-interacting protein(Beclin-1), selective autophagy receptor p62/SQSTM1 and microtubule-associated protein 1 light chain 3 beta(LC3B). Expression change of p62/SQSTM1 was further observed by immunofluorescence staining, and hepatocyte apoptosis was detected by TUNEL assay. Results Hepatocyte swelling and degeneration, inflammatory cell infiltration and bile duct epithelial cell proliferation were observed and serum ALT, AST, ALP, TBA and TBIL levels increased in the ANIT model group, suggesting model successfully established; in the QCLGF intervention group, liver pathological injury was markedly alleviated, and serum biochemical parameters reduced greatly compared to in the model group(P<0.05); hepatic Beclin-1 expression increased, p62/SQSTM1 expression decreased and conversion of LC3B-I to LC3B-II promoted in QCLGF intervention group; in addition, p62/SQSTM1 immunofluorescence intensity was reduced, and percentage of TUNEL-positive cells was decreased after QCLGF intervention. Conclusions QCLGF markedly alleviates ANIT-induced CLI in mice, and the possible protective mechanism might be associated with regulation of Beclin-1, p62/SQSTM1 and LC3B expression, as well as reduction of apoptosis in liver tissues.

Key words: Cholestatic liver injury, α-naphthyl isothiocyanate, Qingchang Ligan Formula, herbal medicine, Autophagy, Mice