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转化生长因子β1诱导皮肤成纤维细胞向肌成纤维细胞表型转分化的机制研究
熊杰,夏照帆*,吕开阳,王钰,韩姝
0 字体:加大+\|默认\|缩小-
(第二军医大学长海医院烧伤科，全军烧伤研究所，上海 200433)

摘要:

目的：探讨TGFβ1诱导皮肤成纤维细胞(FB)向肌成纤维细胞转分化的可能途径和调控机制。方法：将野生型和Smad3基因敲除(KO)型小鼠皮肤FB分为9组：野生型FB组、野生型FB+TGFβ1组、野生型FB+SB431542组、野生型FB+SB431542+TGFβ1组、Smad3 KO FB组、Smad3 KO FB+TGFβ1组、野生型FB+SB203580+TGFβ1组、野生型FB+PD98059+TGFβ1组和野生型FB+SP600125+TGFβ1组。各组细胞经同步化处理后，直接以TGFβ1刺激或经上述各激酶抑制剂预处理后再以TGFβ1刺激。收集细胞，一部分以单细胞RTPCR检测αSMA阳性表达百分比，另一部分细胞抽提总RNA后采用实时荧光定量RTPCR检测αSMA的表达水平变化。结果：Smad3 KO组与SB431542组的αSMA表达水平和阳性百分比显著升高（Smad3 KO FB组vs野生型FB组；野生型FB+SB431542+TGFβ1组vs 野生型FB+SB431542组；Smad3 KO FB+TGFβ1组vs Smad3 KO FB组，P<0.01），而SB203580组和SP600125组中αSMA表达水平和阳性百分比升高的作用则被显著抑制（野生型FB+SB203580+TGFβ1组、野生型FB+SP600125+TGFβ1组vs 野生型FB+TGFβ1组，P<0.05）。结论：在TGFβ1诱导成纤维细胞向肌成纤维细胞的转分化过程中，Smad3途径介导抑制作用，而p38/MAPK、JNK/MAPK途径则介导正向调节作用。

关键词: 转化生长因子β Smad3 成纤维细胞肌成纤维细胞转分化小鼠，基因敲除

DOI：10.3724/SP.J.1008.2007.01175

投稿时间：2007-06-18修订日期：2007-09-29

基金项目:科技部“973”项目子课题（2005CB522603）.

TGF-β1-induced transdifferentiation of derma fibroblasts into myofibroblasts:a study of mechanism

XIONG Jie,XIA Zhao-fan*,,LvKai-yang,WANG Yu,HAN Shu

(Department of Burns,Burn Institute of PLA,Changhai Hospital,Second Military Medical University,Shanghai 200433，China)

Abstract:

Objective：To explore the possible pathways and regulatory mechanism of TGFβ1induced transdifferentiation of derma fibroblasts(FB) into myofibroblasts. Methods: Mice Wildtype and Smad3 knockout(Smad3 KO) derma FB were divided into 9 groups,namely,A:Wildtype FB; B:Wildtype FB+TGFβ1; C:Wildtype FB+SB431542; D:Wildtype FB+SB431542+TGFβ1; E:Smad3 KO FB; F:Smad3 KO FB+TGFβ1; G:Wildtype FB+SB203580+TGFβ1; H:Wildtype FB+PD98059+TGFβ1; and I:Wildtype FB+SP600125+TGFβ1. After synchronization treatment,the cells were treated with TGFβ1 with or without pretreatment with above mentioned kinases inhibitors. Then the cells were collected for RNA extraction and the expression of αSMA was detected by real time quantitative RTPCR; some cells were analyzed by single cell RTPCR to test the positive expression rate of αSMA.Results: The expression and positive rate of αSMA in SB431542 group and Smad3 knockout group were significantly increased（group E vs. group A；group D vs. group C；group F vs. group E，P<0.01） and those in SP600125 group and SB203580 group were significantly inhibited（group G and I vs. group B，P<0.05）. Conclusion: In TGFβ1induced derma fibroblasts transdifferentiation into myofibroblasts,Smad3 pathway plays a negative regulatory role and p38/MAPK and JNK/MAPK pathway play a positive regulatory role.

Key words: transforming growth factor beta Smad3 fibroblast myofibroblast transdifferentiation mice,knockout