胆总管结扎胆汁淤积性肝硬化动物模型的建立与评价  

Establishment and evaluations of cholestatic cirrhosis model induced by bile duct ligation

作　　者：肖程 康权[2] 罗庆[2] Xiao Cheng;Kang Quan;Luo Qing(Second Affiliated Hospital,Army Military Medical University,Xinqiao Hospital,Chongqing,400037;Children's Hospital Affiliated to Chongqing Medical University,Stem Cell Laboratory of Pediatric Research Institute,Key Research Laboratory of Pediatric Developmental Diseases of Ministry of Education,National Key International Laboratory of Children's Developmental Diseases,Chongqing Key Laboratory of Pediatrics,Chongqing 400014)

机构地区：[1]陆军军医大学第二附属医院(新桥医院),重庆市400037 [2]重庆医科大学附属儿童医院,儿科研究所干细胞实验室,儿童发育疾病研究教育部重点实验室,儿童发育重大疾病国家国际科技合作基地,儿科学重庆市重点实验室,重庆市400014

出　　处：《临床小儿外科杂志》2020年第1期74-80,共7页Journal of Clinical Pediatric Surgery

基　　金：国家自然科学基金资助项目(编号:NSFC81172545);重庆市科委基础与前沿研究计划资助项目(编号:cstc2013jcyA10026)

摘　　要：目的探讨胆总管结扎手术致胆汁淤积性肝硬化小鼠模型的建立价值,并对模型进行评价。方法将84只balb/c小鼠随机分为实验组(采用胆总管结扎术)和对照组(采用假手术),术后观察小鼠一般情况和肝胆大体结构;采用全自动生化分析仪检测术后1 d、3 d、5 d、7 d、14 d、21 d、28 d总胆红素(total bilirubin,TBIL)、直接胆红素(direct bilirubin,DBIL)、谷氨酸转氨酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST);采用H&E染色评估肝脏组织形态学变化;采用masson染色和免疫组织化学检测术后28 d小鼠肝脏组织切片,评估两组胶原纤维增生和a-SMA、CK-19蛋白表达情况。结果实验组与对照组小鼠相比,术后一般情况相对更差,胆汁淤积,胆囊、胆总管体积明显增大,各项血清生化指标明显增高(P<0.05);HE染色可见肝小叶结构改变,胆管增生;术后28 d实验组小鼠masson染色可见胶原纤维形成,免疫组织化学染色提示a-SMA、CK-19表达量显著增多(P<0.05)。结论经腹中线小切口行胆总管结扎手术可成功构建小鼠胆汁淤积性肝硬化模型。Objective To explore the establishment and evaluations of cholestatic cirrhosis induced by bile duct ligation in mice.Methods A total of 84 balb/c mice were randomly divided into experimental group(bile duct ligation)and control group(sham operation).General status and hepaticobiliary structures were observed postoperatively.And serum levels of total bilirubin(TBIL),alanine transaminase(ALT),direct bilirubin(DBIL)and aspartate transaminase(AST)were detected by an automatic biochemistry analyzer at Day 1/3/5/7/14/21/28 post-operation.And the histological changes of hepatic tissue were assessed by hematoxylin&eosin(H&E)stain while the hepatic expressions of collagen fiber,a-SMA and CK-19 protein at Day 28 was assessed by Masson's stain and immunohistochemistry for evaluating the hepatic expressions of collagen fiber,a-SMA and CK-19 protein at 28 day post-operation.Results Compared with control group,mice in experimental group had worse general status,cholestasis,gallbladder&common bile duct expansion and cholestasis.Serum biochemical parameters significantly increased(P<0.05).H&E stain showed changes in hepatic lobule and bile duct proliferation.Collagen fiber formation was observed by Masson stain in experimental group at Day 28 day post-operation and immunohistochemical staining indicated that a-SMA and CK-19 increased markedly.Conclusion A murine model of cholestatic cirrhosis is established successfully by bile duct ligation through small incision of abdominal midline.

关 键 词：胆总管 结扎术 肝硬化 胆汁性 模型 动物 

分 类 号：R726[医药卫生—儿科] R575.7[医药卫生—临床医学]