胆总管结扎胆汁淤积性肝硬化动物模型的建立与评价

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。

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

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.