基于CYP7A1/FXR/SHP途径探究大黄素对胆汁淤积性肝损伤小鼠的治疗作用

doi:10.12092/j.issn.1009-2501.2026.02.008

摘要/Abstract

摘要：

目的: 探讨大黄素对胆汁淤积性肝损伤小鼠的治疗作用。方法: 采用炔雌醇（ethinyl estradiol，EE）连续7 d皮下注射C57BL/6J小鼠建立胆汁淤积模型，检测小鼠体质量变化及血清谷丙转氨酶（ALT）、谷草转氨酶（AST）、谷氨酰转肽酶（γ-GT）、总胆红素（TBIL）水平变化，HE（hematoxylin-eosin）染色和MASSON染色评价对肝脏等脏器的影响。通过动物模型给药确定药效后，石胆酸（lithocholic acid，LCA）诱导人正常肝细胞L02构建胆汁淤积细胞模型，采用CCK-8法，通过LCA干预24 h检测半数致死量（IC₅₀）确定造模浓度；通过大黄素干预24 h，检测各组的存活率确定大黄素给药浓度；再检测评价大黄素对LCA造模后肝细胞的存活率。并通过检测各组L02细胞肝损伤指标相关酶ALT、AST水平及氧化应激相关指标谷胱甘肽（GSH）、血清超氧化物歧化酶（SOD）、丙二醛（MDA）和活性氧（ROS）水平的变化，评价大黄素是否通过改善氧化应激发挥抗胆汁淤积的作用机制；Western blot分别检测小鼠肝脏中限速酶-胆固醇7-羟化酶（cholesterol 7α-hydroxylase，CYP7A1）、核受体法尼醇X受体（farnesol X receptor，FXR）和小异二聚体伴侣（SHP）的蛋白表达情况。结果: 大黄素显著改善小鼠体质量变化、肝指数，降低了血清中ALT、AST、γ-GT和TBIL的水平，改善了肝脏的病变趋势，抑制了肝脏损伤。提升了L02细胞存活率，改善了细胞的生存状态；降低了ALT和AST水平的变化，改善了细胞损伤。还通过提升GSH和SOD水平，降低MDA和ROS水平改善了氧化应激反应水平。此外，大黄素明显增加了小鼠肝脏中FXR、SHP并降低了CYP7A1的蛋白表达水平。结论: 大黄素可以对胆汁淤积性肝损伤小鼠起到保护作用，其机制可能是通过抑制氧化应激损伤并通过FXR/CYP7A1/SHP途径调节胆汁酸合成代谢起到保护作用。

关键词: 大黄素, 胆汁淤积, 肝损伤, 氧化应激

Abstract:

AIM: To investigate the therapeutic effect of emodin on cholestatic liver injury in mice. METHODS: C57BL/6J mice were subcutaneously injected with ethinyl estradiol (EE) for 7 consecutive days to establish a cholestasis model. The changes of body weight and serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (γ-GT) and total bilirubin (TBIL) were detected. HE (hematoxylin-eosin) staining and MASSON staining were used to evaluate the effects on liver and other organs. After the efficacy was determined by animal model administration experiment, the human normal liver cell L02 was induced by lithocholic acid (LCA) to construct a cholestatic cell model. The CCK-8 method was used to detect the median lethal dose (IC₅₀) by LCA intervention for 24 hours to determine the modeling concentration. Through emodin intervention for 24 h, the survival rate of each group was detected to determine the concentration of emodin. The survival rate of emodin on liver cells after LCA modeling was detected and evaluated. By detecting the changes of liver injury index related enzymes ALT, AST levels and oxidative stress related enzymes glutathione (GSH), serum superoxide dismutase (SOD), malondialdehyde (MDA) and reactive oxygen species(ROS) levels in each group of L02 cells, the mechanism of emodin in anti-cholestasis by improving oxidative stress was evaluated. Western blot was used to detect the protein expression of Cholesterol 7α-hydroxylase (CYP7A1), Farnesol X receptor (FXR) and small heterodimer partner (SHP) in the liver of mice. RESULTS: Emodin significantly improved the body weight change and liver index of mice, reduced the levels of ALT, AST, γ-GT and TBIL in serum, improved the trend of liver lesions, and inhibited liver damage. Improving the survival rate of L02 cells improved the survival state of cells, reduced the changes of ALT and AST levels and improved cell damage. It also changed the level of oxidative stress by increasing GSH and SOD levels and reducing MDA and ROS levels. In addition, emodin significantly increased FXR and SHP in the liver of mice and decreased the protein expression level of CYP7A1. CONCLUSION: Emodin can play a protective role in cholestatic liver injury, and its mechanism may be through inhibiting oxidative stress injury and regulating bile acid synthesis and metabolism through FXR / CYP7A1 / SHP pathway.

Key words: emodin, cholestasis, liver injury, oxidative stress

中图分类号:

R965.2

王玮辰, 马雅萍, 王萌, 李庆林, 程卉. 基于CYP7A1/FXR/SHP途径探究大黄素对胆汁淤积性肝损伤小鼠的治疗作用[J]. 中国临床药理学与治疗学, 2026, 31(2): 204-212.

Weichen WANG, Yaping MA, Meng WANG, Qinglin LI, Hui CHENG. Therapeutic effect of emodin on cholestatic liver injury in mice based on CYP7A1 / FXR / SHP pathway[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(2): 204-212.

图/表 7

图 1

Fig.1 Effect of emodin on physiological and biochemical indexes of EE model mice ($ \overline{x} $±s, n=6) A-B: effect of body weight and liver index on C57BL/6J mice. C-F: effect of ALT, AST, γ-GT and TBIL levels in C57BL/6J mice. cP<0.01, compared with control group. eP<0.05, fP<0.01, compared with model group.

图 2

Fig.2 Pathological effects of HE staining sections of liver, heart, kidney, spleen and ileum in C57BL/6J mice The black arrow indicates severe inflammatory cell infiltration, liver, heart, kidney, spleen (×100), ileum(×200).

图 3

Fig.3 Pathological effects of MASSON staining sections of C57BL/6J mouse liver ($ \overline{x} $±s, n=6) The black arrow indicates severe inflammatory cell infiltration (×100).

图 4

Fig.4 Effect of emodin on LCA-induced L02 cell viability ($ \overline{x} $±s, n=6) A: effect of LCA on cytotoxicity. B: effect of emodin on cytotoxicity. C: effect of emodin on LCA-induced cell viability. bP<0.05, cP<0.01, compared with control group; eP<0.05,fP<0.01, compared with model group.

图 5

Fig.5 Effect of emodin on ALT and AST in LCA-induced L02 cells ($ \overline{x} $±s, n=3) cP<0.01, compared with control group; eP<0.05, fP<0.01, compared with model group.

图 6

Fig.6 Effect of emodin on GSH, SOD, MDA and ROS in LCA-induced L02 cells ($ \overline{x} $±s, n=3) cP<0.01, compared with control group; eP<0.05, fP<0.01, compared with model group.

图 7

Fig.7 Effect of emodin on protein expression of FXR, CYP7A1and SHP induced by LCA in CLD mouse ($ \overline{x} $±s, n=3) cP<0.01, compared with control group.

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