加味茵陈蒿汤对α⁃萘异硫氰酸酯诱导的胆汁淤积小鼠肝损伤作用机制研究  

Study on mechanism of modified Yinchenhao Decoction on liver injury induced byα⁃naphthyl isothiocyanate in mice with cholestasis

作　　者：徐韵 陈影[2] 许媛媛 蔡静雯 叶青艳[1] 陈建杰[1] 凌琪华[1] 赵钢[1] XU Yun;CHEN Ying;XU Yuanyuan;CAI Jingwen;YE Qingyan;CHEN Jianjie;LING Qihua;ZHAO Gang(Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Ruijin Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China;Shanghai Medical Group Joint Management and Development Center,Shanghai 200001,China)

机构地区：[1]上海中医药大学附属曙光医院,上海201203 [2]上海交通大学医学院附属瑞金医院,上海200025 [3]上海市医事团体联合管理发展中心,上海200001

出　　处：《上海中医药大学学报》2023年第6期41-48,共8页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(82004321);上海市“科技创新行动计划”医学创新研究专项项目(21Y11920500);上海市进一步加快中医药传承创新发展三年行动计划项目(ZY〔2021-2023〕-0101-02)。

摘　　要：目的:探讨加味茵陈蒿汤治疗α-萘异硫氰酸酯(ANIT)诱导的胆汁淤积小鼠肝损伤的作用机制。方法:24只雄性SPF级C57BL/6小鼠随机分为正常对照组(Normal组)、ANIT造模组(ANIT组)、加味茵陈蒿汤组(JWYCH组)、熊去氧胆酸组(UDCA组),每组6只。Normal组与ANIT组灌胃给予饮用水,持续10 d;JWYCH组灌胃给予加味茵陈蒿颗粒剂水溶液,UDCA组灌胃给予熊去氧胆酸悬液,持续10 d。第7天,小鼠用ANIT油溶液(50 mg/kg)造模。末次给药后,采用全自动生化仪检测小鼠血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、总胆汁酸(TBA)及总胆红素(TBiL)水平;苏木素-伊红(HE)染色观察肝组织形态学变化;采用RT-qPCR检测肝组织法尼醇X受体(FXR)、成纤维细胞生长因子15(FGF15)、FGF受体4(FGFR4)mRNA表达;Western blot检测肝组织FGF15、FXR、磷酸化FXR(p-FXR)、FGFR4、外排型转运体[胆盐输出泵(BSEP)、多药耐药相关蛋白2(MRP2)]、摄取型转运体[钠离子-牛磺胆酸共转运蛋白(NTCP)、有机阴离子转运多肽2(OATP2)]蛋白表达。结果:血清肝功能生化学指标显示,与Normal组相比,ANIT组ALT、ALP、AST、TBA、TBiL水平明显升高(P<0.05);JWYCH组和UDCA组ALT、ALP、AST、TBA、TBiL水平明显低于ANIT组(P<0.05);与UDCA组相比,JWYCH组的TBA水平明显下降(P<0.05)。肝组织病理结果显示,光镜下ANIT组可见胆管内胆栓形成,其汇管区以及胆管周围可见大量炎性细胞浸润,提示造模成功;JWYCH组、UDCA组病理损伤较ANIT组有明显改善。Western blot结果显示,与Normal组比较,ANIT组FGF15、FXR、p-PXR、FGFR4、BSEP、MRP2蛋白表达均明显减少(P<0.05),提示造模成功;与ANIT组比较,JWYCH组和UDCA组FGF15、FXR、p-PXR、FGFR4、BSEP、MRP2蛋白表达明显增加(P<0.05,P<0.01),而NTCP、OATP2表达明显减少(P<0.05)。RT-qPCR结果显示,与Normal组比较,ANIT组小鼠肝组织FXR、FGF15、FGFR4的mRNA表达明显降低(P<0.05);与ANIT组比较,UDCA组和JWYCH�Objective:To investigate the mechanism of modified Yinchenhao Decoction(JWYCH)in the treatment ofα-naphthyl isothiocyanate(ANIT)-induced hepatic injury in mice with cholestasis.Methods:Twenty-four male SPF grade C57BL/6 mice were randomly divided into blank group(Normal group),ANIT modeling group(ANIT group),JWYCH group and ursodeoxycholic acid group(UDCA group),with 6 mice in each group.Normal and ANIT groups were given drinking water by gavage for 10 d,JWYCH and UDCA groups were given JWYCH granules aqueous solution and ursodeoxycholic acid by gavage for 10 d,respectively.On the 7th day,mouse model were set up by ANIT oil solution(50mg/kg).After the last administration,the serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),total bile acid(TBA)and total bilirubin(TBiL)were detected by automatic biochemical analyzer;Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of liver;Farnisol X receptor(FXR),fibroblast growth factor 15(FGF15)and FGF receptor 4(FGFR4)mRNA expressions were detected by RT-qPCR;protein expressions of FGF15,FXR,phosphorylated FXR(p-FXR),FGFR4,effluent transporters[bile salt export pump(BSEP),multidrug resistance associated Protein 2(MRP2)],uptake transporters[sodium-taurocholic acid cotransporter(NTCP),organic anion transporter polypeptide 2(OATP2)]were detected by Western blot.Results:①Serum liver function indexes showed that compared with the Normal group,the levels of ALT,ALP,AST,TBA and TBiL in the ANIT group were significantly increased(P<0.05),the levels of ALT,ALP,AST,TBA and TBiL in JWYCH and UDCA groups were significantly lower than those in the ANIT group(P<0.05);Compared with the UDCA group,TBA level in the JWYCH group was significantly decreased(P<0.05).②The histopathological results showed that the formation of bile duct thrombi was observed in the ANIT group under light microscopy,and a large number of inflammatory cells could be seen in the portal area and around the bile duct,indicating succe

关 键 词：加味茵陈蒿汤 原发性胆汁性胆管炎 胆汁淤积 肝损伤 

分 类 号：R285.5[医药卫生—中药学]