高糖对大鼠系膜细胞肝细胞生长因子受体表达的影响及其机制研究  

Impact of high glucose on hepatocyte growth factor receptor expression in rat mesangial cells

作　　者：李慧[1] 蒋涛[1] 林伊凤[1] 赵仲华[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：《中华肾脏病杂志》2007年第6期372-376,共5页Chinese Journal of Nephrology

基　　金：国家自然科学基金（30570857）

摘　　要：目的观察高糖作用下大鼠系膜细胞（MsC）肝细胞生长因子（HGF）受体c-Met的表达,并探讨其机制和意义。方法用RT-PCR和Western印迹方法检测高糖作用大鼠MsC的不同时间点（0、12、24、48、96h）c-Met的表达。分别用光辉霉素A（mithramycin A）和SU11274抑制转录因子Sp1的DNA结合活性和阻断c-Met。用电泳迁移率改变实验（EMSA）观察Sp1与c-Met基因启动子的结合活性。以荧光探剂二氯双氢荧光素二乙酸酯（DCFH-DA）捕获细胞内活性氧。结果大鼠MsC的c-Met表达在高糖作用12、24和48h都明显上升,96 h开始下降。光辉霉素A呈浓度依赖性抑制高糖作用下大鼠MsC的c-Met表达上调。大鼠MsC内Sp1与c-Met基因启动子的结合活性在高糖作用下明显增强。HGF及c-Met显著抑制高糖诱导的大鼠MsC内活性氧的增多。结论高糖作用下大鼠MsC的c-Met表达增强,其机制可能是通过Sp1介导。HGF-c-Met信号通路激活能抑制高糖所致大鼠MsC内的氧化应激反应。Objective To observe hepatocyte growth factor receptor c-Met expression in high glucose-treated rat mesangial cells （MsC） and explore its significance and relevant mechanism. Methods RT-PCR and Western blot analysis were performed to detect c-Met expression in MsC cultured under high glucose conditions for different time （0, 12, 24, 48, 96 h）. Spl-binding inhibitor mithramycin A and c-Met inhibitor SU11274 were used to block Spl binding DNA and c-Met respectively. With electrophoretic mobility shift assay （EMSA）, binding of Sp1 to c-Met promoter was evaluated. Intracellular reactive oxygen species was examined by loading MsC with dichlorodihydrofluorescein diacetate, a redox-sensitive fluorescent dye. Results Exposure MsC to high glucose resulted in an increase in c-Met mRNA and protein expression at first 48 h then a following decrease at 96 h. Mithramycin A suppressed high glucose-increased c-Met expression in a dose-dependent manner. Enhanced Spl binding to c-Met promoter was shown in high glucosetreated MsC. HGF/c-Met markedly reduced intracellular reactive oxygen species in MsC cultured in high glucose. Conclusions High glucose may activate c-Met expression in MsC through Sp1. HGF/c-Met can attenuate high glucose-mediated oxidative stress.

关 键 词：原癌基因蛋白质C-MET 肝细胞生长因子 糖尿病肾病 系膜细胞 转录因子SP1 氧化应激 

分 类 号：R686[医药卫生—骨科学]