胆汁淤积性肝硬化大鼠模型的改良

摘要

Objective To establish a rat model of liver fibrosis induced by bile duct ligation ( BDL ) , and to improve the classical BDL method .Methods Eighty adult healthy male Sprague-Dawley rats were randomly divided into groups A and B , 40 rats each .The two groups of rats were modeling by classical BDL method and modified hepatic duct ligation method , respectively .Sham operation was performed in 10 rats of each group and were taken as control sham group.Hepatic biochemical indicators AST , ALT, ALP, TBIL, DBIL, GGT, and A/G in the rat serum at one week after operation were detected with an automatic biochemistry analyzer .Pathological changes of the rat liver at 4 weeks after operation were observed with HE staining .Cell proliferation-related markers α-SMA and CK-19 in the liver were detected by immunohistochemistry .Results Both two rat models of obstructive cholestasis were successfully established .Serum biochemical and liver pathological changes indicated impaired liver function in the bile duct ligated rats .After cholestasis caused by bile duct ligation , mitotic figures of hepatocytes and the expression of α-SMA and CK-19 in hepatocytes and biliary epithelial cells was increased , and hyperplasia of bile canaliculi and hepatic pseudolobule formation were observed . These changes indicated early hepatic cirrhosis .The mortality rate of rats was 66.7%in the BDL group and 26.7%in the modified BDL group.Conclusions Liver fibrosis can be successfully induced by hepatic duct ligation in rats .In addition, this modified ligation method can reduce the mortality and improve the model quality and efficacy of experiment .%目的构建肝纤维化大鼠模型，并对经典结扎胆总管（ BDL）复制模型方法进行适当改进。方法80只SD雄性成熟大鼠，按随机数字表法分为A组40只、B组40只，分别运用胆总管结扎法和肝门部肝总管缝扎法先后两次造模，各取其中10只为假手术组进行对照，术后1周眼眶取血采用酶联免疫吸附法检测血清谷草转氨酶（AST）、谷丙转氨酶（ALT）、碱性磷酸酶（ALP）、总胆红素（TBIL）、直接胆红素（DBIL）、γ-谷氨酰转肽酶（GGT）、白蛋白/球蛋白（ A/G），术后4周HE染色观察大鼠肝组织病理学变化，采用免疫组化染色分析肝脏组织α-SMA和CK-19表达水平。结果两种方法均表现出明显肝功能损害；标本胆小管增生明显，肝脏假小叶形成，达到早期肝硬化，肝脏α-SMA和CK-19表达水平明显升高，胆总管结扎组死亡率为66.7％，肝门部肝总管缝扎组死亡率为26.7％。结论肝门缝扎法可成功建立胆汁淤积性肝硬化大鼠模型，能够明显降低模型动物死亡率，提高模型质量及实验效率。