目的:通过观察胆汁酸转运蛋白胆盐输出泵(BSEP)、钠离子-牛磺胆酸共转运蛋白(NTCP)在熊果酸(UA)干预酒精肝损害(ALD)小鼠后的表达情况,研讨UA对ALD小鼠的保护作用。方法:10周龄SPF级C57BL/6J小鼠18只,随机数字表法分为3组(正常对照组、酒精模型组以及UA+酒精组),连续灌胃干预7周,记录小鼠体质量变化,HE染色法观察肝组织病理学变化,微板法检测血清总胆汁酸(TBA)、谷草转氨酶(AST)、谷丙转氨酶(ALT),Western blotting检测肝组织BSEP、NTCP蛋白表达量。结果:酒精模型组小鼠平均体质量较正常对照组明显降低(P＜0.05);正常对照组小鼠肝小叶的结构完整,无炎性细胞浸润,而酒精模型组肝小叶结构模糊,且伴有明显的脂肪变性,NAS评分显著增高(P＜0.05);与酒精模型组相比,经UA干预后,小鼠肝小叶结构损伤有所改善、肝细胞脂肪空泡与炎性细胞数目减少,NAS评分明显降低(P＜0.05)。此外,与正常对照组相比,酒精模型组小鼠肝脏指数显著增高(P＜0.05),而UA+酒精组小鼠肝脏指数较酒精模型组明显降低差异(P＜0.05)。同时,经UA干预后,可明显抑制因酒精摄入诱发的TBA、AST与ALT含量升高(P＜0.05)。Western blotting检测结果显示,与正常对照组相比,酒精模型组小鼠肝组织中转运蛋白BSEP、NTCP表达水平明显降低(P＜0.05);经UA干预后,UA+酒精组小鼠转运蛋白BSEP、NTCP蛋白表达水平明显升高(P＜0.05)。结论:UA的补充可以明显的改善因酒精所致的肝脏损害,作用机理可能与其上调BSEP、NTCP蛋白表达,进而调节胆汁酸稳态有关。
Objective: To investigate the protective effect of ursolic acid (UA) on alcoholic liver damage (ALD) mice by observing the expression of bile salt export pump (BSEP) and sodium taurocholate cotransporting polypeptide (NTCP) in mice after UA intervention. Methods: Eighteen 10-week-old SPF C57BL/6J mice were randomly divided into three groups (normal control group, alcohol model group, and UA+alcohol group). The mice were treated with intragastric administration for 7 weeks. The changes of body weight were recorded. The pathological changes of liver tissue were observed by HE staining. The serum total bile acid (TBA), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were detected by microplate method. The expression of BSEP and NTCP protein in liver tissue was detected by Western blotting. Results: The average body weight of mice in the alcohol group was significantly lower than that in the normal control group (P＜0.05). The structure of hepatic lobules in the normal control group was intact and there was no inflammatory cell infiltration, while the structure of hepatic lobules in the alcohol model group was blurred, accompanied by obvious steatosis, and the NAS score was significantly increased (P＜0.05). Compared with the alcohol model group, after UA intervention, the damage of hepatic lobule structure in mice was improved, the number of fat vacuoles and inflammatory cells in hepatocytes was reduced, and the NAS score was significantly reduced (P＜0.05). In addition, compared with the normal control group, the liver index of the alcohol model group was significantly increased (P＜0.05), while the liver index of the UA+alcohol group was significantly lower than that of the alcohol model group (P＜0.05). At the same time, after UA intervention, the increase of TBA, AST and ALT induced by alcohol intake was significantly inhibited (P＜0.05). The results of Western blotting showed that compared with the normal control group, the expression levels of BSEP and NTCP in the liver tissue of the alcohol model group were significantly decreased (P＜0.05). After UA intervention, the expression levels of transporter BSEP and NTCP protein in UA+alcohol group were significantly increased (P＜0.05). Conclusion: UA supplementation can significantly improve liver damage caused by alcohol, and the mechanism may be related to the up-regulation of BSEP and NTCP protein expression, thereby regulating bile acid homeostasis.
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