骨形成蛋白-7对肝纤维化表皮生长因子受体表达的影响  

Effect of bone morphogenetic protein-7 intervention on epidermal growth factor receptor expression in liver fibrosis

作　　者：王丽萍[1] 董进中[1] 曹肃婷[1] 张赛男[1] 林镯[1] 王晓东[1] 陈永平[1] 

机构地区：[1]温州医科大学附属第一医院感染科,325000

出　　处：《中华传染病杂志》2014年第2期89-93,共5页Chinese Journal of Infectious Diseases

基　　金：王宝恩肝纤维化研究基金（xjs001）;“艾滋病和病毒性肝炎等重大传染病防治”科技重大专项--中医药辨证论治方案阻断、逆转乙型肝炎相关肝纤维化的临床疗效评价研究“十二五”（2013ZX10005002-001）

摘　　要：目的观察肝纤维化小鼠模型中表皮生长因子受体（EGFR）的动态表达，以及骨形成蛋白-7（BMP-7）对其表达的影响，探讨治疗纤维化的新靶点。方法30只健康雄性ICR小鼠随机数字表法分为正常对照组（6只）、肝纤维化模型组（18只，包括4、8、12周3个亚组）和BMP-7干预组（6只），皮下注射四氯化碳制备肝纤维化小鼠模型，BMP-7干预组注射四氯化碳8周后腹腔注射人重组BMP-7，持续4周。采用HE和Manson染色观察肝组织病理变化，并进行肝纤维化半定量评分；下腔静脉采血检测ALT、AST和白蛋白；采用实时荧光定量PCR和Western印迹法分别检测各组TGF-β1 mRNA和TGF-β1、EGFR、pEGFR蛋白表达情况。计量资料比较采用方差分析，相关性采用Pearson相关分析。结果成功建立小鼠肝纤维化模型，肝纤维化模型组血清ALT、AST均逐渐升高，12周时最高，白蛋白逐渐降低，12周时最低，BMP-7干预组各指标均有所缓解[ALT：（153．9±18．1）U／L比（191．3±24．5）U／L；AST：（177．8±19．2）U／L比（206．6±25．0）U／L；白蛋白：（25．4±0．9）g／L比（22．2±1．2）g／L；均P〈0．053。肝纤维化组随着纤维化的进展，TGF-β1、EGFR及pEGFR的蛋白表达量均逐渐升高，于12周时达到高峰，BMP-7干预后，上述3种蛋白的表达水平均明显降低（TGF-β1：0．256士0．006比0．287±0．014；EGFR：1．061±0．017比1．094±0．014；pEGFR：0．855±0．053比1．007±0．063；均P〈0．05）。直线相关分析显示，TGF-β1与EGFR及pEGFR表达量均呈正相关（rs值分别为0．895、0．859，均P〈0．05）。结论BMP-7能改善小鼠肝纤维化，其作用机制可能是调节EGFR和TGF-β1的表达。Objective To investigate the dynamic expressions of epidermal growth factor receptor （EGFR） in mice with liver fibrosis and the effect of bone morphogenetic protein-7 （BMP-7） intervention on the expression of EGFR, and to explore a new therapy target for fibrosis. Methods A total of 30 healthy male ICR mice were randomly divided into three groups.. 6 mice in control group, 18 mice in hepatic fibrosis group and 6 mice in BMP-7 intervention group. The model of mice with liver fibrosis was established by subcutaneous injection of carbon tetrachloride （CC14） for 12 weeks. After administration of CC14 for 8 weeks, human recombinant BMP-7 was given into mice in intervention group by intraperitoneal injection for 4 weeks. Hematoxylin-Eosin and Masson staining of liver tissues were employed to observe the pathological changes, and the semi-quantitative analysis of liver fibrosis was performed. Blood withdrawn from inferior vena cava was detected for levels of alanine aminotransferase （ALT）, aspartate aminotransferase （AST） and albumin （Alb）. The expressions of transforming growth factor-β1 （TGF-β1） mRNA and TGF-β1, EGFR, phosphorylation EGFR （pEGFR） protein in each group were detected using quantitative real-time polymerase chain reaction and Western blot. Measurement date was compared using analysis of variance and Pearson correlation analysis. Results The model of mice with liver fibrosis was successfully established. In model group, the serum levels of ALT and AST increased, while the level of Alb decreased gradually. All these biochemical index improved after intervention of BMP-7 （ALT： [153.9±18.1] U/L vs [191.3±24.5] U/L;AST：[177.8±19. 2] U/L vs [206.6±25.0] U/L;Alb： [25.4±0.9] g/L vs [22.2±1.2] g/L; all P〈0.05）. With the progress of fibrosis, TGF-β1, EGFR and pEGFR protein expressions increased gradually in model group and reached a peak at week 12, which was significantly different compared to the control group （all P〈0.05）. In BMP-7 intervention group,

关 键 词：肝硬化 骨形态发生蛋白质类 受体 表皮生长因子 转化生长因子B1 疾病模型 动物 

分 类 号：R575.2[医药卫生—消化系统]