高糖对脑微血管内皮细胞抗氧化酶系统的影响  

Effects of Hyperglycemia on Antioxidant Enzymes in Brain Microvessel Endothelial Cells

作　　者：陈峰[1] 刘志民[2] 赵瑛[1] 

机构地区：[1]第二军医大学长征医院神经内科,上海200003 [2]第二军医大学长征医院内分泌科,上海200003

出　　处：《标记免疫分析与临床》2010年第5期309-312,共4页Labeled Immunoassays and Clinical Medicine

摘　　要：探讨高糖刺激时脑微血管内皮细胞锰超氧化物歧化酶及其他抗氧化酶的基因表达改变。通过体外培养小鼠脑微血管内皮细胞系bEnd.3;H2DCF荧光法检测高糖刺激下脑微血管内皮细胞短期内氧自由基(ROS)生成,RT-PCR检测高糖刺激下内皮细胞铜锌超氧化物歧化酶(SOD1)、锰超氧化物岐化酶(MnSOD)、脂质过氧化物酶(GPX)及触酶(CAT)的表达,并通过werstern blot法检测内皮细胞内MnSOD蛋白水平。结果表明:高糖刺激下脑微血管内皮细胞短期内氧自由基(ROS)产生明显增多并持续增高;高糖刺激10小时抗氧化酶MnSOD,GPX,CAT的mRNA水平无明显变化,SOD1 mRNA水平略有增高;SOD1、MnSOD的mRNA水平在高糖刺激24小时后有明显增加,但是MnSOD蛋白水平在高糖刺激48小时内并无明显增加;抗氧化剂白藜芦醇或表没食子儿茶素没食子酸酯(EGCG)不能上调MnSOD蛋白水平。以上数据表明,高糖刺激能诱导细胞内多种抗氧化酶包括MnSOD基因表达上调,但也能抑制MnSOD蛋白水平的增加。To explore the effects of hyperglycemia on MnSOD and other antioxidant enzymes in brain microvessel endothelial cells under hyperglycemic conditions,mouse brain microvessel endothelial cells line bEnd.3 was used.Reactive oxygen species（ROS） production was measured using 2＇7＇-dichlorofluorescin diacetate（DCFH-DA）;SOD1,MnSOD,GPX and CAT gene expression were determined in bEnd.3 under high glucose（25mM） or normal glucose（5.5mM） by reverse transcription PCR.The protein level of MnSOD and the effect of Resveratrol or epigallocatechin gallate（EGCG） on protein level of MnSOD were also determined.The results showed that hyperglycemia can significantly increased the mRNA levels of MnSOD and CuZnSOD,while there was no increase in protein levels of MnSOD under hyperglycemia condition.Resveratrol and EGCG can not be used to increase the MnSOD protein levels.The present study suggests that hyperglycemia can inhibit the increase of MnSOD protein level which maybe exacerbates oxidative stress.

关 键 词：糖尿病 脑微血管内皮细胞 锰超氧化物歧化酶 氧化应激 

分 类 号：R743.3[医药卫生—神经病学与精神病学]