肝纤维化模型中Nogo-参与转化生长因子-β1／Smad2信号通路的研究  

Study on Nogo-B participating in transforming growth factor-β1/Smad2 signaling pathway in mice modelsof hepatic fibrosis

作　　者：刘真真[1] 陈永平[1] 沈宇娟[1] 王雅琴[1] 宋梅[1] 林镯[1] 

机构地区：[1]温州医学院附属第一医院感染科,325000

出　　处：《中华传染病杂志》2013年第5期269-273,共5页Chinese Journal of Infectious Diseases

摘　　要：目的探讨在小鼠肝纤维化模型中Nogo-B与转化生长因子β1（TGF-β1）／Smad2信号通路的关系。方法将24只健康雄性ICR小鼠分为正常对照组6只和肝纤维化模型组18只，模型组再按造模后不同时间点分为4、8和12周3个亚组。肝纤维化模型组皮下注射四氯化碳（CCl4）诱导小鼠肝纤维化模型，肝组织经HE和Masson染色后，光学显微镜下观察各组肝组织病理变化；RTPCR方法检测各组肝脏Nogo-B、Smad2和TGF-β1 mRNA表达；Western印迹法、免疫组织化学法检测各组肝脏NogoB、Smad2、TGF-β1蛋白表达。组间均数比较采用单因素方差分析。结果CCl4成功诱导肝纤维化模型。Nogo-B两种亚型Nogo-B1、Nogo-B2 mRNA在正常肝组织表达为0．140±0．050和0．104±0．023，在肝纤维化模型组肝脏中表达显著增加，造模12周时为1．054±0．040和0．500±0．057（F=431．41、135．46，均P〈0．01）；Nogo-B蛋白主要表达于肝脏间质中，肝细胞仅少量表达；且Nogo-B mRNA及蛋白的表达强度与参与肝纤维化信号通路相关因子Smad2、TGF-β1mRNA及蛋白均呈正相关（均P〈0．01）。结论NogoB可能通过参与TGF-β1／Smad2信号通路在肝纤维化发展过程中起重要作用，促进肝纤维化的发展。Objective To study the relationship between Nogo-B and transforming growth factor-β1 （TGF-β1）/Smad2 signaling pathway in mice models of hepatic fibrosis. Methods Twenty four healthy male ICR mice were divided into two groups, with 6 in the control group and 18 in the model group. Mice in the model group were further divided into three subgroups according to different time points： subgroups of 4, 8 and 12 weeks, with 6 mice in each subgroup. Hepatic fibrosis of mice was induced by subcutaneous injection of carbon tetrachloride （CCl4）. The histopathologic changes of the liver were observed by optical microscope using hematoxylin eosin and Masson trichrome stainings of the liver tissues. Expressions of Nogo-B, Smad2 and TGF-β1 mRNA and proteins in liver were detected by reverse transcription-polymerase chain reaction （ RT PCR ）, Western blot and immunohistochemistry assays, respectively. Means among groups were compared by univariate analysis of variance. Results The hepatic fibrosis models were successfully induced by CC14 injection. The expressions of two subtypes of Nogo B, Nogo-B1 and Nogo-B2 mRNA in normal livers were 0. 140±0. 050 and 0. 104±0. 023, but both significantly increased in the livers of mice in the 12 weekmodel subgroup （1. 054±0. 040 and 0. 500±0. 057, F=431.41 and 135.46, respectively; both P〈 0.01）. The Nogo-B protein was mainly expressed in nonparenchymal cells of the liver, and was hardly expressed in hepatocytes. Linear correlation analysis showed that the expressions of Nogo B mRNA and proteins were positively correlated with Smad2 and TGF-β1 mRNA and proteins （all P〈0.01）, which were considered to participate in the signaling pathway of hepatic fibrosis. Conclusion Nogo-B might play a role in the development and progression of hepatic fibrosis by participating in TGF-β1/ Smad2 signaling pathway.

关 键 词：髓磷脂蛋白质类 转化生子因子β1 肝硬化 DNA结合蛋白质类 疾病模型 动 物 动力传导 细胞 

分 类 号：R575[医药卫生—消化系统] R-332[医药卫生—内科学]