转化生长因子β1对乳鼠心肌细胞转化生长因子结合蛋白2表达的影响  

Effect of transforming growth factor beta 1 on neonatal rat latent transforming growth factor beta binding protein 2 in cardiomyocytes

作　　者：白媛媛[1] 魏英杰[1] 崔传珏[1] 李君[1] 刘晓艳[1] 史强[1] 

机构地区：[1]北京协和医学院中国医学科学院阜外心血管病医院心血管病研究所心血管疾病国家重点实验室,北京100037

出　　处：《基础医学与临床》2012年第2期143-148,共6页Basic and Clinical Medicine

基　　金：国家重点基础性研究项目(973项目)(2010CB529506);"重大新药创制"科技重大专项-"创新药物临床研究技术平台建设"(2008ZX09312-018);教育部高等学校博士学科点专项科研基金(20091106110018)

摘　　要：目的探讨转化生长因子β1(TGF-β1)在诱导心肌细胞表达转化生长因子结合蛋白2(LTBP2)中的作用及信号传导通路。方法培养乳鼠心肌细胞;实时定量聚合酶链式反应、蛋白质印迹和免疫细胞化学方法检测不同时间和不同浓度的TGF-β1对大鼠乳鼠心肌细胞LTBP2基因及蛋白表达的影响;用TGF-β1相关信号通路阻断剂探讨TGF-β1调节LTBP2表达改变的信号传导机制。结果 LTBP2基因表达随着TGF-β1浓度增加(0、2、5及10μg/L)而明显升高,在5μg/L时刺激最强(P<0.05);5μg/L的TGF-β1刺激下心肌细胞内LTBP2基因和蛋白表达的升高呈时间依赖性,均在12 h最高,24 h开始呈下降趋势(P<0.05或P<0.01);免疫细胞化学结果显示TGF-β1明显升高LTBP2的表达。信号传导通路研究显示TGF-β1在心肌细胞内主要通过ERK信号通路和PI3K信号通路诱导LTBP2的表达。结论 TGF-β1在乳鼠心肌细胞内通过ERK信号通路和PI3K信号通路上调LTBP2的表达。Objective To explore transforming growth factor beta 1 （TGF-β1） induced latent transforming growth factor beta binding protein 2 in neonatal rat cardiomyocytes with its underlying signal transduction pathway. Meth- ods Primary cultured eardiomyocytes were obtained from neonatal rats by enzymatic digestion method. LTBP2 gene and its protein expression were examined by real-time PCR, Western blot analysis and immunocytochemistry. In or- der to further study the signal transduction pathway of TGF-β1 to inhibit the effect of TGF-β1 and to observe the expression c , inhibitors of TGF-β1 signaling pathways were used hange of LTBP2. Results The stimulation effect of TGF-β1 was dose-dependent, LTBP2 gene expression began to increase after 2 ug/L （P 〈 0. 05 ） of stimulation, peaked at 5 ug/L （P 〈 0. 05） and reached a plateau after 10 ug/L （P 〈 0.05 ）. The stimulation effect of TGF-β1 was also time - dependent, with the stimulation time extension, the expression of LTBP 2 was increased, peaked at12 h and declined at 24 h （P 〈 0. 05 ）. Immunocytochemistry results showed that TGF-β1 increased LTBP2 levels. Signaling pathway study demonstrated that TGF-131 induced expression of LTBP2 in cardiomyocytes was via ERK pathway and PI3K pathway. Conclusion TGF-β1 upregulates LTBP2 expression in cardiomyocytes via ERK path- way and PI3K pathway.

关 键 词：转化生长因子Β1 心肌细胞 转化生长因子结合蛋白2 

分 类 号：R541[医药卫生—心血管疾病]