高糖对大鼠肾系膜细胞Iκ B-α表达影响的泛素化研究  

Expression of IκB-α in cultured rat glomerular mesangial cell induced by high glucose via the ubiquitin-proteasome pathway

作　　者：徐玲[1] 蒋岚[1] 马红艳[1] 徐勇[1] 杨军[1] 

机构地区：[1]泸州医学院附属医院内分泌代谢科,四川泸州646000

出　　处：《中国现代医学杂志》2010年第24期3691-3693,3697,共4页China Journal of Modern Medicine

基　　金：国家自然科学基金资助项目(No:30670980);四川省杰出青年基金项目(08ZQ026-366)

摘　　要：目的探讨高糖对大鼠肾小球系膜细胞Iκ B-α表达的影响及其可能的作用途径。方法将大鼠HBZY-1肾小球系膜细胞进行体外培养,设立正常对照组(5.6 mmol/L葡萄糖)、高糖组(包括10、20和30mmol/L葡萄糖)、甘露醇组、30 mmol/L高糖加特异性泛素蛋白酶体阻断剂MG132组及5.6 mmol/L葡萄糖加MG132组,作用48 h,用Western Blotting法检测各组IκB-α蛋白的表达,实时定量PCR法检测各高糖组IκB-α及泛素mRNA的表达。结果各高糖组Iκ B-α蛋白的表达下降(P<0.05),具有浓度依赖效应;各高糖组Iκ B-αmRNA的表达差异无统计学意义(P>0.05);各高糖组泛素mRNA的表达增加(P<0.05);30mmol/L高糖加MG132组较30 mmol/L高糖组Iκ B-α蛋白的表达下降被明显逆转(P<0.01)。结论高糖可增加大鼠肾系膜细胞泛素表达,上调泛素蛋白酶体系统活性,增加Iκ B-α蛋白泛素化降解。【Objective】To study the effect of the expression of IκB-α and the role of ubiquitin-proteasome system in cultured Rat Glomerular Mesangial Cells（GMC） induced by high glucose.【Methods】Cultured HBZY-1 rat GMC were divided into 7 groups： normal glucose（5.6 mmol/L） group,high glucose（10,20,30 mmol/L） group,mannito（24.4 mmol/L） group,30 mmol/L glucose plus MG132 group,（MG132,a proteasome inhibitor）,5.6 mmol/L glucose plus MG132 group.The expression of IκB-α protein were measured by Western-Blotting,the expression of IκB-α mRNA and ubiquitin mRNA were measured by real-time Quantitative PCR.【Results】①Compared with normal glucose group,the expression of IκB-α protein was decreased significantly in a dose dependent manner in high glucose groups（P 0.05）.② The expression of IκB-α mRNA had no significant change in each group（P 0.05）.③ Compared with normal glucose group,the expression of ubiquitin mRNA was increased obviously in high glucose group（P 0.05）.④ Compared with 30 mmol/L high glucose groups,the expression of IκB-α protein could be reverted mostly by adding the MG132（P 0.01）.【Conclusions】High glucose can increase the expression of ubiquitin mRNA and upregulate ubiquitin-proteasome system,increase the degradation of IκB-α in cultured GMC,which is independent of the osmotic pressure.

关 键 词：糖尿病肾病 高糖 肾小球系膜细胞 Iκ B-α 泛素 

分 类 号：R587.1[医药卫生—内分泌]