Smad泛素化调节因子2在肝纤维化过程中对结缔组织生长因子的作用研究  

The effect of Smad ubiquitination regulatory factor 2 on connective tissue growth factor during liver fibrosis

作　　者：蔡瑜[1] 徐奕[2] 沈锡中[1] 

机构地区：[1]复旦大学附属中山医院消化科,上海200032 [2]复旦大学附属中山医院护理部,上海200032

出　　处：《内科理论与实践》2012年第4期305-309,共5页Journal of Internal Medicine Concepts & Practice

基　　金：上海市自然科学基金(项目编号:12ZR1405300)

摘　　要：目的:在体内和体外研究Smad泛素化调节因子2(Smurf2)在肝纤维化过程中对结缔组织生长因子(CTGF)的作用。方法:建立四氯化碳诱导肝纤维化大鼠模型,收集人类肝硬化和正常肝组织样本,用逆转录-聚合酶链反应(RT-PCR)、蛋白质印迹(Western blot,WB)、免疫组织化学(IHC)方法检测Smurf2和CTGF的表达;培养人类肝星状细胞(LX-2),构建Smurf2表达质粒和小干扰RNA(siRNA),在转化生长因子β1(TGF-β1)作用后,通过WB检测Smurf2过表达和小干扰后对CTGF表达的影响。结果:在大鼠肝纤维化时,Smurf2表达增加,伴有CTGF水平增高;大鼠和人类肝硬化组织中,Smurf2表达反而减少,CTGF表达仍然增加。体外肝星状细胞用TGF-β1处理后,CTGF蛋白水平升高;过表达Smurf2可以阻断CTGF蛋白的合成;抑制Smurf2表达后,CTGF表达持续增加。TGF-β1处理LX-2后可以诱导Smurf2表达内源性轻度增加。结论:Smurf2可以由TGF-β1诱导产生,作为负反馈抑制子抑制肝纤维化过程中CTGF表达的增加。Objective To study the effect of Smad ubiquitination regulatory factor 2 （Smurf 2） on connective tissue growth factor （CTGF） during liver fibrosis. Methods CC14 induced rat liver fibrosis model was constructed, and fibrosis and cirrhosis liver samples in rat and normal and cirrhosis liver samples in human were collected. Levels of Smurf 2 and CTGF expression were assessed by reverse transcription polymerase chain reaction 0RT-PCR）, Western blotting （WB）and immuno- histochemistry （ IHC）. Human hepatic stellate cell line, LX-2, were cultured. Smurf 2 expression vector and small interfering RNA （siRNA）were constructed. After treated with transforming growth factor -β1 （TGF-β1）, the effect of overexpression of Smurf 2 and siRNA on expression of CTGF were examined by WB. Results The increase of Smurf 2 level was accompanied with increasing in CTGF levels in liver fibrosis stage. In cirrhosis stage, however, CTGF expression increased continuously while Smurf 2 expression was decreased. CTGF level in LX-2 increased after TGF-β1 treatment. Overexpression of Smurf 2 could inhibit the synthesis of CTGF; inhibition of Sumrf2 expression induced continuous increase of CTGF. TGF-β1 treatment could induce mild intrinsic increase of Smurf 2 expression. Conclusions Smurf 2 expression could be induced by TGF-β1 and act as a negative-feedback inhibitor of CTGF expression during the process of liver fibrosis.

关 键 词：Smad泛素化调节因子2 结缔组织生长因子 肝纤维化 泛素蛋白水解途径 转化生长因子Β 

分 类 号：R575.2[医药卫生—消化系统]