醛糖还原酶(AR)基因对大鼠肾系膜细胞的转染及AR抑制剂对其增殖的影响  

Aldose Reductase(AR) Gene Transfection and AR Inhibitors on the Proliferation of Rat Mesangial Cells

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：《复旦学报（医学版）》2005年第4期383-387,共5页Fudan University Journal of Medical Sciences

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)基因转染及AR抑制剂(ARI)对体外培养大鼠肾系膜细胞(MsC)增殖的影响。方法应用Westernblot检测转基因MsC及PDGF作用MsC后AR的表达。四唑蓝比色法(MTT法)检测细胞增殖,流式细胞术分析细胞周期,对比大鼠正常MsC和转基因MsC的生长情况,并观察ARI———Sorbinil和Zopolrestat对血小板源性生长因子(PDGF)和小牛血清(NBS)促细胞增殖作用的影响。结果①Westernblot鉴定转基因MsCAR表达较正常MsC明显增高;②转染组较正常组生长速度快;③PDGF作用MsC后可上调AR的表达;④PDGF和10%NBS均可显著刺激两组细胞的增殖,ARI可部分抑制PDGF对正常组和转染组的促增殖作用,对10%NBS,ARI可部分抑制其对转染组的促增殖作用,而对正常组无显著影响,抑制作用呈剂量依赖性。结论AR可能参与了MsC在病理状况下的过度增殖过程。Purpose To study the effects of aldose reductase (AR) gene transfection and AR inhibitors (ARI) on the proliferation of rat mesangial cells (MsC) in vitro. Methods The efficiency of transfection and the effects of platelet-derived growth factor (PDGF) on the expression of AR was assessed by Western blot.Cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)colorimetric assay.Cell cycle was analyzed by flow cytometry.The proliferation of normal MsC and transfected MsC used AR were compared.It were verified whether ARI--Sorbinil and Zopolrestat,could affect the proliferation induced by PDGF and 10% new bovine serum (NBS). Results The AR expression of transfected MsC was obviously higher than normal MsC.The growth of transfected MsC was quicker than normal MsC.Meanwhile,PDGF upregulated the AR expression of transfected and normal MsC.ARI could partially inhibit the proliferation stimulated by PDGF and 10%NBS except for the 10%NBS towards normal MsC. Conclusions AR might participate in the regulation of MsC proliferation in pathologenesis.

关 键 词：醛糖还原酶(AR) 大鼠肾系膜细胞 抑制剂 WESTERNBLOT 血小板源性生长因子 促细胞增殖作用 促增殖作用 PDGF BLOT检测 MsC 流式细胞术 剂量依赖性 转基因 ARI 正常组 体外培养 基因转染 MTT法 细胞周期 小牛血清 生长速度 

分 类 号：R284.1[医药卫生—中药学] R692.3[医药卫生—中医学]