结缔组织生长因子增加TGFβ活化成肌纤维细胞的作用  

CTGF enhances TGFβ_1-mediated myofibroblast activation

作　　者：杨敏[1] 黄海长[1] 李惊子[1] 王海燕[1] 

机构地区：[1]北京大学第一医院肾脏内科暨肾脏病研究所,北京100034

出　　处：《基础医学与临床》2005年第1期25-29,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金 (30 2 70 6 12 );教育部教育振兴行动计划专项基金 (985工程 );教育部留学回国人员科研启动基金

摘　　要：探讨结缔组织生长因子 (CTGF)对转化生长因子 β1(TGFβ1)诱导成肌纤维细胞的影响及其与Smad信号通路的关系。细胞免疫组化检测成纤维细胞表型转化。Western免疫印迹法检测细胞α 平滑肌肌动蛋白 (α-SMA)及磷酸化Smad2的水平 ,免疫共沉淀检测Smad2 / 3与Smad4复合物的水平 ,提取核蛋白后再用Western免疫印迹法检测Smad2在细胞核内的聚积。结果显示CTGF不能使细胞表达α-SMA ,TGFβ1显著促进α- SMA表达 (p <0.0 1) ,CTGF加TGFβ1共同刺激又明显高于TGFβ1(p <0. 0 5 )。TGFβ1可明显诱导Smad2磷酸化、Smad2 / 3,4复合物形成以及Smad2在细胞核内的聚集 ,CTGF加TGFβ1联合刺激与TGFβ1相比没有明显变化。提示CTGF可以增强TGFβ1诱导的成肌纤维细胞生成 。To investigate whether connective tissue growth factor (CTGF) has an effect on transforming growth factor-β 1 (TGFβ 1)-mediated myofibroblastic activation, and on the Smad signaling pathway which accounts for TGFβ 1's action. Immunocytochemistry was used for the assay of the production of myofibroblasts, α-smooth muscle actin and phospho-Smad2 protein level was detected by Western-blot. Immunoprecipitation was performed to measure the association of Smad2/3 with Smad4. Nuclear protein was isolated and then subjected to detect activated Smad2 nuclear accumulation. CTGF had no prominent effect on the induction of myofibroblast and expression of α-SMA in renal interstitial fibroblast NRK-49F cells, whereas TGFβ1 significantly stimulated myofibroblastic activation and the expression of α-SMA (P<0.05). Simultaneous incubation of CTGF enhanced TGFβ 1-induced myofibroblast and de novo α-SMA expression significantly(P<0.05). CTGF neither affected Smad2 phosphorylation and their association with Smad4, nor affect activated Smad2 nucleus accumulation in cells. It suggests that CTGF synergistically enhanced TGFβ 1-innitiated myofibroblastic activation in renal interstitial fibroblasts, this action seems to be independent of the potentiation on the cascade of TGFβ 1-triggered Smad signal transduction.

关 键 词：TGFΒ1 SMAD2 CTGF 成肌纤维细胞 Α-SMA 结缔组织生长因子 诱导 细胞核 磷酸化 信号传导 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]