蒿芩清胆汤调控枯否细胞M1/M2极化对原发性胆汁性胆管炎的机制研究  

Experimental study of regulating M1/M2 polarization of Kupffer cell by Haoqin Qingdan Decoction to improve intrahepatic bile duct injury in the treatment of primary biliary cholangitis

作　　者：徐俊 郭晟 周莉涵 尹谢添 谭张奎 XU Jun;GUO Sheng;ZHOU Li-han(College of Basic Medicine,Hubei University of Traditional Chinese Medicine(Wuhan Hubei,430065),China.;不详)

机构地区：[1]湖北中医药大学基础医学院,湖北武汉430065 [2]湖北时珍实验室 [3]湖北中医药大学附属湖北省中医院

出　　处：《中西医结合肝病杂志》2024年第12期1096-1100,共5页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：湖北省自然科学基金联合项目(No.2024AFD328)。

摘　　要：目的:基于枯否细胞M1/M2极化探讨蒿芩清胆汤对原发性胆汁性胆管炎(PBC)肝内胆管的保护作用及机制。方法:60只小鼠随机分为对照组、模型组、蒿芩清胆汤(HQQD)不同剂量组、熊去氧胆酸(UDCA)组,利用2-辛炔酸-牛血清白蛋白(2-OA-BSA)联合聚肌胞苷酸诱导的PBC小鼠模型,相应药物灌胃干预12周。苏木精伊红(HE)染色法观察肝组织病理学改变,全自动生化分析仪检测血清γ-谷氨酰转肽酶(γ-GT)、总胆汁酸(TBA)、总胆红素(TBil)、天门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)改变,实时荧光定量(RT-PCR)检测一氧化氮合成酶(iNOS)、肿瘤坏死因子α(TNF-α)、甘露糖受体1(CD206)、白介素10(IL-10)mRNA的表达变化,蛋白免疫印记法(WB)检测iNOS、TNF-α、CD206、IL-10蛋白表达变化情况。结果:病理学结果显示,对照组小鼠肝组织门管区结构清晰,小叶间胆管周围无异常改变,模型组小鼠门管区可见多发炎症细胞聚集,炎症细胞聚集范围内可见肉芽肿、淋巴细胞浸润,HQQD高、中、低剂量组与UDCA组门管区炎症细胞数量较模型组减轻。生化分析结果显示,与对照组相比,模型组小鼠血清γ-GT、TBA、TBil、AST、ALT水平均不同程度升高;与模型组比较,UDCA组和HQQD高、中、低剂量组小鼠血清γ-GT、TBA、TBil、AST、ALT表达均明显下调。RT-PCR和WB结果显示,与对照组相比,模型组小鼠肝组织iNOS、TNF-αmRNA及蛋白表达水平明显上调,CD206、IL-10 mRNA及蛋白表达明显下调;与模型组比较,UDCA组和HQQD高、中、低剂量组iNOS、TNF-αmRNA及蛋白表达水平明显下调,CD206、IL-10 mRNA及蛋白表达水平明显上调。结论:蒿芩清胆汤可以使PBC模型小鼠枯否细胞M1极化减弱,M2极化增强,调控M1/M2极化状态,改善胆汁淤积及肝内胆管损伤。Objective:Exploring the protective effects and mechanisms of Haoqin Qingdan Decoction on intrahepatic bile ducts in primary biliary cholangitis(PBC)via regulation of Kupffer cell M1/M2 polarization.Methods:60 mice were randomly divided into control group,model group,Haoqinqingdan Decoction(HQQD)high-dose,medium-dose and low-dose groups,ursodeoxycholic acid(UDCA)group,the PBC mouse model induced by 2-octyneic acid and bovine serum albumin(2-OA-BSA)in combination with polyinosine acid(PolyI:C)was subjected to treatment with the corresponding drug for a duration of 12 weeks.The liver histopathological changes were observed by HE staining,the serumγ-GT,TBA,Tbil,AST,ALT were detected by automatic biochemical analyzer,and the expression of iNOS,TNF-α,CD206 and IL-10 mRNA were detected by RT-PCR.The expression of iNOS,TNF-α,CD206 and IL-10 were detected by WB.Results:The pathological results indicated that the hepatic tissue in the control group exhibited a clear portal area structure without any abnormal changes around the interlobular bile ducts.In the model group,multiple inflammatory cell clusters were observed in the portal area,accompanied by granuloma formation and lymphocyte infiltration.Compared to the model group,the number of inflammatory cells in the portal area was reduced in the high-medium-,and low-dose HQQD groups as well as in the UDCA group.The biochemical analysis results showed that,compared to the control group,the serum levels ofγ-GT,TBA,TBil,AST,and ALT were elevated to varying degrees in the model group.In contrast,the UDCA group and the high-,medium-,and low-dose HQQD groups exhibited significantly decreased serum levels ofγ-GT,TBA,TBil,AST,and ALT compared to the model group.The RT-PCR and Western blot(WB)results demonstrated that,compared to the control group,the hepatic expression levels of iNOS and TNF-αmRNA and proteins were significantly upregulated in the model group,while the expression levels of CD206 and IL-10 mRNA and proteins were significantly downregulated.However,in the UDCA

关 键 词：蒿芩清胆汤 原发性胆汁性胆管炎 肝内胆管损伤 枯否细胞极化 

分 类 号：R259[医药卫生—中西医结合]