目的 探讨茵栀黄颗粒治疗小鼠胆汁淤积症的作用及其机制。方法 将8周龄C57BL/6小鼠给予含1%胆酸的饲料2周，构建小鼠胆汁淤积症模型，将小鼠分为对照组、模型组、熊去氧胆酸（0.1 g/kg）组以及茵栀黄颗粒（5、10、20 g/kg）组，各组分别ig相应药物，对照组和模型组大鼠ig同体积生理盐水，连续给药4周。分别给予收集小鼠血清，全自动生化仪测定肝功能相关指标，超高效液相色谱串联质谱（UPLC-MS/MS）测定胆汁酸成分；收集小鼠肝组织，苏木精–伊红（HE）染色观察肝脏组织病理变化；RT-qPCR检测肝组织炎症因子和胆汁酸代谢关键基因的mRNA表达水平；Western blotting检测肝组织胆汁酸代谢关键蛋白的表达情况；收集小鼠盲肠内容物，Illumina Miseq测序平台对V₃～V₄可变区进行扩增和测序，对小鼠肠道菌群结构进行分析。结果 与模型组相比，茵栀黄颗粒各组小鼠血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、碱性磷酸酶（ALP）水平均显著降低（P＜0.05、0.01、0.001）；且能够显著改善小鼠肝组织损伤。与模型组相比，茵栀黄颗粒各剂量组小鼠肝脏白细胞介素-10（IL-10）、肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、单核细胞趋化蛋白-1（MCP-1）mRNA表达水平均显著降低，白细胞介素-1β（IL-1β）、胆固醇7-羟化酶（CYP7A1）、胆盐输出泵（Bsep）、牛磺胆酸钠共转运蛋白（Ntcp）、胆汁酸转运蛋白多药耐药相关蛋白2（Mrp2）、UDP葡糖醛酸基转移酶1家族多肽A1（Ugt1a1）mRNA表达水平均显著升高（P＜0.05、0.01、0.001），并影响胆汁酸代谢关键蛋白的mRNA和蛋白表达。16S rDNA测序显示，茵栀黄颗粒组小鼠肠道菌群中乳杆菌属相对丰度显著升高，罗斯氏菌属和Allobaculum属相对丰度显著降低。胆汁酸代谢组学发现，茵栀黄颗粒能够降低多种次级胆汁酸的含量。结论 茵栀黄颗粒改善胆汁淤积的作用机制可能与调控肠道菌群组成影响胆汁酸代谢有关。

Objective To investigate the effect and mechanism of Yinzhihuang Granules in treatment of cholestasis mice. Methods C57BL/6 mice at the age of 8 weeks were given a diet containing 1% cholic acid for 2 weeks to establish a mouse cholecystasis model, and the mice were divided into control group, model group, ursodeoxycholic acid (0.1 g/kg) group, and Yinzhihuang Granules (5, 10, 20 g/kg) groups, each group was given the corresponding drug intragaically, and the rats in the control group and model group were given the same volume of normal saline intragaically. The drug was administered continuously for 4 weeks. Serum was collected, liver function indexes were determined by automatic biochemical analyzer, and bile acid components were determined by UPLC-MS/MS. Liver tissues of mice were collected and stained with hematoxylin-eosin (HE) to observe the pathological changes of liver tissue. The mRNA expression levels of inflammatory factors and key genes of bile acid metabolism were detected by RT-qPCR. The expression of key proteins of bile acid metabolism in liver tissues was detected by Western blotting. The contents of mouse cecum were collected, and the V₃ — V₄ variable regions were amplified and sequenced by Illumina Miseq sequencing platform, and the structure of mouse intestinal flora was analyzed. Results Compared with model group, the serum levels of ALT, AST, and ALP in Yinzhihuang Granules group were significantly decreased (P < 0.05, 0.01, 0.001), and it can significantly improve the liver tissue damage of mice. Compared with model group, the mRNA expression levels of liver IL-10, TNF-α, IL-6, and MCP-1 in each dose group of Yinzhihuang Granules were significantly decreased. The mRNA expression levels of IL-1β, CYP7A1, Bsep, Ntcp, Mrp2, and Ugt1a1 were all positive significantly increased (P < 0.05, 0.01, 0.001), and affected the mRNA and protein expression of key proteins in bile acid metabolism. 16S rDNA sequencing showed that the abundance of Lactobacillus in gut microbiota of mice in Yinzhihuang Granules group was significantly increased, and the abundance of Roseburia and Allobaculum was significantly reduced. Bile acid metabolomics found that Yinzhihuang Granules could reduce the content of various secondary bile acid. Conclusion Mechanism of Yinzhihuang Granules on improving cholestasis liver injury may be related to regulating the composition of gut microbiota and affecting bile acid metabolism.

国家自然科学基金青年基金资助项目（81900468）；河南省医学科技攻关计划联合共建项目（LHGJ20220419）；郑州大学第一附属医院横向课题（K2019-0148）