醛糖还原酶基因转染对体外培养大鼠肾脏系膜细胞增殖的影响  

Effects of aldose reductase transfection on the proliferation of rat mesangial cells in vitro

作　　者：车祺[1] 蒋涛[1] 林伊凤[1] 李慧[1] 张农[1] 

机构地区：[1]复旦大学上海医学院病理学系,上海200032

出　　处：《中华病理学杂志》2005年第7期417-420,共4页Chinese Journal of Pathology

基　　金：国家自然科学基金资助项目(30170430)

摘　　要：目的观察醛糖还原酶(AR)基因转染对体外培养大鼠肾脏系膜细胞(MsC)增殖的影响,并对其机制进行初步探讨。方法脂质体转染法将AR基因转入大鼠MsC,蛋白印迹鉴定转染效率。四甲基偶氮唑盐(MTT)比色法检测细胞增殖,流式细胞术分析细胞周期与凋亡,对比正常MsC和AR转染MsC的生长速度及AR抑制剂(ARI)Sorbinil和Zopolrestat对血小板源性生长因子-BB(PDGF-BB)和小牛血清(NBS)促MsC增殖作用的影响。蛋白印迹检测PDGF-BB作用后AR和c-Jun表达的变化,EMSA检测转录因子AP-1的活性。结果(1)转染MsCAR表达较正常MsC明显增高;(2)AR转染MsC较正常MsC生长速度快(P<0.01);(3)ARI可部分抑制PDGF-BB对正常MsC、AR转染MsC以及NBS对AR转染MsC的促增殖作用(P<0.01,P<0.05),而对NBS促正常MsC的增殖无显著影响;(4)PDGF-BB刺激MsC后可上调AR和c-Jun蛋白的表达,ARI可减弱c-Jun的这种表达增高;(5)PDGF-BB可活化转录因子AP-1,而ARI可削弱此作用。结论AR可能参与了MsC在病理情况下的过度增殖过程,其作用机制与AP-1活化有一定关系。Objective To study the effects of aldose reductase (AR) on the proliferation of rat mesangial cells (MsC) in vitro and to investigate its mechanism. Methods Cell proliferation was assessed by MTT colorimetric assay. Cell cycle and apoptosis were analyzed by flow cytometry. The growth of normal MsC and AR transfected MsC was compared. The proliferation of PDGF-BB and cellular growth stimulation by 10% NBS were investigated using AR inhibitors (ARI) Sorbinil and Zopolrestat. The effects of PDGF-BB on the expression of AR,p65 and c-Jun were assessed by Western blot. Activation of AP-1 was measured by EMSA.Results AR expression of transfected MsC was distinctly higher than that of the control. Transfected MsC grew quicker than normal cells. ARI partially inhibited the proliferation of transfected MsC under the stimulation of PDGF-BB and 10%NBS, whereas 10%NBS had no effect on normal MsC. PDGF-BB upregulated the expression of AR and c-Jun, but had no effect on p65. The upregulation of c-Jun and the activation of AP-1 could be attenuated by ARI. Conclusion AR may participate in the pathological proliferation of MsC through the pathway related to the activation of AP-1.

关 键 词：体外培养 基因转染 系膜细胞增殖 大鼠 转录因子AP-1 PDGF-BB 醛糖还原酶(AR) 血小板源性生长因子 四甲基偶氮唑盐 c-Jun表达 c-Jun蛋白 肾脏系膜细胞 脂质体转染法 AP-1活化 MsC 蛋白印迹 生长速度 流式细胞术 促增殖作用 

分 类 号：R692[医药卫生—泌尿科学]