过表达TRAP-1-LikeProtein基因对人皮肤成纤维细胞的影响  

Effects of TRAP-l-like protein gene on human dermal fibroblasts

作　　者：王雪[1] 陈骏[1] 钱云良[1] 沃雁[1] 王琛[1] 王丹茹[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院整复外科,200011

出　　处：《中华实验外科杂志》2012年第11期2144-2147,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30400467）;上海市科委基金资助项目（09ZRl416900）

摘　　要：目的通过在人皮肤成纤维细胞（HSFs）中过表达TRAP-1-LikeProtein（TLP），观察其对细胞生物学性状的影响。方法通过慢病毒途径在人正常皮肤成纤维细胞中获得稳定过表达TLP基因后，检测细胞增殖、收缩活性以及TLP、Ⅰ、Ⅲ型胶原的表达变化。结果慢病毒介导外源性TLP基因在细胞中稳定表达。TLP可协同转化生长因子-β1（TGF-β1）促进细胞活力升高60％（48h）（P〈0．05）；纤维凝胶体积变化差异有统计学意义（P〈0．05）；实时定量聚合酶链反应（Real-timePCR）和Westernblot结果都显示L／Ⅲ型胶原的合成与TLP的表达量呈正相关（P〈0．05），细胞因子TGF-β1刺激下此反应更明显。结论TLP可促进人成纤维细胞增殖和收缩，并促进其合成Ⅰ／Ⅲ型胶原。Objective To investigate the biological effects of overexpression of TRAP-1-like protein （TLP） on human dermal fibroblasts. Methods Lentiviral vectors encoding TLP was constructed to transfect fibroblasts derived from normal human skin. After stable transfection, the variation of Cell viability, contraction activity and the expression levels of TLP and COL Ⅰ/Ⅲ genes in various groups induced by transforming growth factor-β1（TGF-β1） after TLP gene treatment were detected. Results Exogenetic TLP gene delivery system mediated by lentivirus vector could be expressed stably in cells. The cell viability of skin fibroblasts was increased by up to 60% after TLP treatment cooperated with TGF-β1 （ P 〈 0. 05 ）. TLP could facilitate the contraction of collagen gel induced by fibroblasts down to 70% of its previous vol- ume, Real-time PCR and Western blotting revealed that the synthesis of COL Ⅰ/Ⅲ was positively correlated with the expression of TLP. These variation trends became obvious under the stimulation of TGF-~I.Con- clusion TLP was able to improve the overall cell viability of skin fibroblasts, as well as, to promote scar contracture. And the synthesis of COL Ⅰ/Ⅲ was positively correlated with the expression of TLP, especially under the stimulation of TGF-β1.

关 键 词：成纤维细胞 胶原 转化生长因子-Β1 

分 类 号：R285.5[医药卫生—中药学]