RKIP在胆总管结扎肝纤维化大鼠肝脏组织表达.docVIP

RKIP在胆总管结扎肝纤维化大鼠肝脏组织的表达 　　作者：马俊骥 姜慧卿 李芳芳 崔东来 刘 丽 扈彩霞 张杰英2 作者单位：(河北医科大学第二医院消化内科，河北 石家庄 050000) 　　【摘要】 目的 研究RKIP在胆总管结扎肝纤维化大鼠肝脏组织中的表达。方法 SD大鼠60只，随机分为模型组(48只)、对照组(12只)。运用胆总管结扎方法制作大鼠肝纤维化模型，模型组分别于结扎后1、2、3、4 w麻醉动物，假手术组与4 w模型组同批麻醉，留取肝脏标本。组织切片经HE和Masson三色染色检测病理变化，Western印迹和免疫组织化学方法检测RKIP在肝组织的表达，用Western印迹方法检测RKIP和ERK的磷酸化水平。结果 HE和Masson三色染色结果证明胆总管结扎大鼠肝纤维化模型制作成功。随着肝纤维化程度增高RKIP表达下降，RKIP和ERK的磷酸化水平则相应增加。结论 胆总管结扎大鼠肝纤维化过程中肝脏组织Raf1/MEK/ERK1,2信号转导途径的激活与RKIP的蛋白表达下降和磷酸化增加有关。 　　【关键词】 RKIP;ERK;肝纤维化 　　【Abstract】 Objective To study Raf kinase inhibitor protein (RKIP) expression in liver of hepatic fibrosis rats induced by common bile duct ligation. Methods Hepatic fibrosis model was induced in SpragueDawley rats by common bile duct ligation (BDL). Livers in model group were harvested at 1, 2, 3 and 4 w after operation. Livers in sham operation group were harvested at 4 w after operation. Histopathological changes were evaluated by hematoxylin and eosin (HE) staining and by Masson′s trichrome method. RKIP expression and phosphorylation of RKIP and ERK in the livers were determined by Western blot, while the distribution of RKIP in the livers was assessed by immunohistochemistry. Results HE and Masson′s trichrome staining showed the successfully established hepatic fibrosis rats by common bile duct ligation. RKIP expression was decreased following the changes of hepatic fibrosis. On the contrary, phosphorylation of RKIP and ERK in the livers was increased. Conclusions The activation of Raf1/MEK/ERK1,2 signaling pathway is correlated with decreased RKIP expression and increased phosphorylated RKIP in the livers of bile duct ligation rats. 　　【Key words】 RKIP; ERK; Hepatic fibrosis 　　Raf激酶抑制蛋白(Raf kinase inhibitor protein，RKIP) 是Yeung等〔1〕发现的一种在信号通路Raf1/MEK/ERK1,2中起负向调节作用的因子。研究证明，肝纤维化(HF)的发生发展与Raf1/MEK/ERK1,2信号通路的激活有密切的关系〔2，3〕，但是RKIP作为此信号通路的抑制因子在肝纤维化进程中的作用机制目前尚未见报道。本实验采用Western印迹法和免疫组化法，研究RKIP在肝脏组织的表达情况，旨在为阐明肝纤维化形成的分子机制提供一定的理论依据。 　　1 材料与方法 　　1.1 动物 选用8周龄健康雄性SD大鼠