逆转素对胆管结扎诱导大鼠肝损害的影响  

Effect of reversine on liver damage induced by bile duct ligation in rats

作　　者：黄迪[1,2,3] 黄宇 黄子圣[1] 翁杰锋 李佩霖 张帅[1,2,3] 古维立 HUANG Di;HUANG Yu;HUANG Zisheng;WENG Jiefeng;LI Peilin;ZHANG Shuai;GU Weili(Department of General Surgery,Guangzhou First People s Hospital,Guangzhou 510180,China;Department of General Surgery,Guangzhou First People s Hospital,School of Medicine,South China University of Technology,Guangzhou 510180,China;Guangzhou Digestive Disease Center,Guangzhou 510180,China)

机构地区：[1]广州市第一人民医院普通外科,广州510180 [2]华南理工大学附属第二医院普通外科,广州510180 [3]广州消化疾病中心,广州510180

出　　处：《广州医药》2021年第3期9-17,共9页Guangzhou Medical Journal

基　　金：贵州省中医药民族医药科技研究项目(QZYY-2020-013);广州市医药卫生科技一般引导项目(20171A011025,20171A011248,20181A011002);广州市中医药科技项目(20172A011004);广东省卫生厅项目(A2020624);广州市第一人民医院红棉项目(M2019003)。

摘　　要：目的探究小分子化合物逆转素(reversine,Rev)对胆管结扎(BDL)诱导的大鼠胆汁淤积性肝损害、纤维化、上皮细胞-间充质转化以及胆管反应的影响。方法雄性Lewis大鼠随机分成三组,每组各5只。按照如下处理:BDL组大鼠行2周的胆管结扎;BDL+Rev组行胆管结扎同时给予腹腔注射逆转素;对照采用假手术(Sham)。2周后获取血液和肝组织。血指标检测总白蛋白(TP)、总胆红素(TBIL)、谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、γ谷氨酰转肽酶(GGT)。H&E染色检测肝组织病理。Azan染色检测组织胶原蛋白。免疫组化检测肝组织α平滑肌肌动蛋白(α-SMA)、结蛋白(Desmin)、波形蛋白(Vimentin)、细胞角蛋白(CK7,CK19)、β-连环蛋白(β-Catenin)以及上皮细胞粘附分子(EpCAM)蛋白的表达情况。结果胆管结扎导致肝脏合成的总白蛋白量下降,总胆红素(TBIL)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、γ谷氨酰转肽酶(GGT)水平明显上升,逆转素处理使下降的总白蛋白上升,使上升的总胆红素(TBIL)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、γ谷氨酰转肽酶(GGT)水平向正常水平回复。逆转素可以缓解胆汁淤积引起的肝纤维化,表现为下调BDL引起的胶原蛋白和α-SMA蛋白沉积。逆转素可以抑制胆汁淤积引起的上皮细胞-间充质转化表现为逆转素明显降低BDL导致的Desmin和Vimentin的表达。逆转素可以抑制胆汁淤积引起的胆管反应表现为明显减少CK7和CK19阳性胆管的表达含量。逆转素抑制胆汁淤积引起的胆管反应与调节β-Catenin和EpCAM的表达有关。结论逆转素可以缓解胆汁淤积引起的大鼠肝损害,具有一定的保护作用。逆转素可以成为一种潜在治疗药物。Objective To investigate the effect of reversine(REV)on bile duct ligation(BDL)-induced hepatic damage,fibrosis,epithelial-mesenchymal transformation,and ductular reaction in rats.Methods Male Lewis rats were randomly divided into three groups with 5 rats in each group.Bile duct ligation was performed in the BDL group for two weeks.BDL+REV group was treated with bile duct ligation and intraperitoneal injection of reversine.The control group was Sham operation(Sham).Blood and liver tissue were obtained after 2 weeks.Blood indexes were determined for total albumin,total bilirubin,alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase andγ-glutamyl transpeptidase.Hepatic histopathology was detected by H&E staining.Azan staining was used to detect tissue collagen deposition.Immunohistochemistry was used to detect the expression ofα-SMA,desmin,vimentin,cytokeratin,β-catenin and epithelial cell adhesion molecule(EpCAM)protein.Results Bile duct ligation resulted in the decrease of total albumin synthesis in liver,and the increase of total bilirubin,glutamic-oxalacetic transaminase,alkaline phosphatase andγ-glutamyltranspeptidase.The levels of total bilirubin,aspartate transaminase,alkaline phosphatase andγ-glutamyltranspeptidase returned to the normal level with reversine treatment.Reversine could alleviate cholestasis-induced liver fibrosis by downregulating BDL-induced deposition of collagen andα-SMA protein.Reversine inhibited cholestasis-induced epithelial-mesenchymal transformation by significantly reducing BDL-induced desmin and vimentin expression.Reversine could inhibit cholestasis-induced ductular reaction by significantly reducing the expression of CK7 and CK19 positive biliary cells.Inhibition of cholestasis induced ductular reaction by reversine was associated with regulation ofβ-catenin and EpCAM expression.Conclusion Reversine can alleviate liver damage caused by cholestasis in rats and have a protective effect.Reversine may be a potential treatment that need further investigation.

关 键 词：逆转素 胆管结扎 胆汁淤积 胆管反应 肝纤维化 

分 类 号：R575.2[医药卫生—消化系统]