清肠利肝方对ANIT诱导的胆汁淤积性肝损伤小鼠保护作用机制研究*

doi:10.3969/j.issn.1672-5069.2026.05.004

摘要/Abstract

摘要： 目的探讨清肠利肝方(QCLGF)对α-萘异硫氰酸酯(ANIT)诱导的小鼠胆汁淤积性肝损伤(CLI)的保护作用及其可能的机制。方法将12只小鼠随机分为对照组、ANIT模型组和QCLGF干预组,每组4只。采用ANIT灌胃法建立小鼠CLI模型,并给予QCLGF干预。采用Western blot法检测小鼠肝组织自噬相关蛋白肌球蛋白样BCL2结合蛋白(Beclin-1)、选择性自噬受体p62(p62/SQSTM1)和微管相关蛋白1轻链3β(LC3B)表达水平,采用p62/SQSTM1免疫荧光染色法观察蛋白表达变化,采用TUNEL法检测小鼠肝细胞凋亡。结果 ANIT模型组小鼠肝细胞肿胀变性、炎症细胞浸润及胆管上皮细胞增生,血清ALT、AST、ALP、TBA和TBIL水平升高,而QCLGF干预组小鼠肝组织病理损伤明显减轻,血清指标显著降低(P < 0.05); QCLGF干预组小鼠肝组织Beclin-1表达显著强于模型组,而p62/SQSTM1表达显著减弱;模型组细胞凋亡率显著高于对照组,而QCLGF干预组细胞凋亡率显著低于模型组(P < 0.05)。结论 QCLGF能够明显减轻ANIT诱导的小鼠CLI,其作用机制可能与调节Beclin-1、p62/SQSTM1和LC3B等自噬相关蛋白表达,并减少肝组织细胞凋亡有关。

关键词: 胆汁淤积性肝损伤, α-萘异硫氰酸酯, 清肠利肝方, 自噬, 小鼠

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

吴小雪, 王昕钰, 胡鸿阳, 李秀惠, 张向颖. 清肠利肝方对ANIT诱导的胆汁淤积性肝损伤小鼠保护作用机制研究^*[J]. 实用肝脏病杂志, 2026, 29(5): 653-656.

Wu Xiaoxue, Wang Xinyu, Hu Hongyang, et al. Protective mechanism of Qingchang Ligan Formula in mice with ANIT-induced cholestatic liver injury[J]. Journal of Practical Hepatology, 2026, 29(5): 653-656.

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清肠利肝方对ANIT诱导的胆汁淤积性肝损伤小鼠保护作用机制研究^*

Protective mechanism of Qingchang Ligan Formula in mice with ANIT-induced cholestatic liver injury

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