波动性高糖对大鼠肾小球系膜细胞氧化应激的影响  

Effects of intermittent high glucose on oxidative stress in rat glomerular mesangial cells in vitro

作　　者：田晓玲[1] 郑景晨[1] 章海凌[1] 沈飞霞[1] 刘芳[1] 

机构地区：[1]温州医学院第一附属医院内分泌科,浙江温州325027

出　　处：《温州医学院学报》2008年第5期422-424,共3页Journal of Wenzhou Medical College

摘　　要：目的:对比研究波动性与稳定性高糖对肾小球系膜细胞(GMC)氧化应激的影响。方法:体外培养大鼠肾小球系膜细胞株,分正常糖对照组(5.5mmol/L,NG)、稳定高糖组(25mmol/L,HG)、波动性高糖组(5.5mmol/L或25mmol/L,每24h交替,IHG),培养GMC6d,分别以二氢二氯荧光素(DCFH-DA)标记细胞,通过流式细胞仪检测细胞内二氯荧光黄(DCF)的荧光强度而测得细胞内ROS水平,比色法检测细胞上清液中的总超氧化物歧化酶(T-SOD)活性和谷胱甘肽(GSH)、丙二醛(MDA)含量的变化。结果:与NG组相比,HG组与IHG组细胞内DCF平均荧光强度均显著升高(均P<0.01),总SOD活力均下降(均P<0.01),GSH含量均下降(P<0.05,P<0.01),MDA均升高(P<0.05,P<0.01)。与HG组相比,IHG组细胞内DCF平均荧光强度显著升高(P<0.01),总SOD活力下降(P<0.01),GSH含量下降(P<0.01),MDA升高(P<0.05)。结论:波动性高糖较稳定性高糖可能对GMC有更强的氧化损伤效应。Objective：To compare the effects of intermittent and constant high glucose on oxidative stress in cultured Rat Glomerular Mesangial Cells （GMC）. Methods： GMC were cultured in normal glucose control group （5.5 mmol/l glucose, NG）, high glucose group （25 mmol/l glucose, HG） and intermittent high glucose group （5.5mmol/1 alternating with 25mmol/l glucose on a daily basis, IHG） in vitro for 6 days. 2＇, 7＇ -dichlorofluorescein diacetate （DCFH-DA） was used as a reactive oxygen species（ROS） capture agent. The fluorescent intensity of 2＇, 7＇ -dichlorofluorescein （DCF）, which was the product of cellular oxidation of DCFH-DA, was detected by flow cytometry, and the level of ROS was thus measured. The activity of total superoxide dismutase （T-SOD） and the levels of malondialdehyde （MDA）, glutathione （GSH）in supernatant were assayed using chromatometry. Results： Compared with NG group, the intracellular fluorescent intensity of DCF increased in HG group and IHG group （P〈 0.01, respectively）, and the activity of T-SOD decreased （P〈 0.01, respectively）, the level of GSH decreased （P〈 0.05, P〈 0.01）, the level of GSH increased （P〈 0.05, P〈 0.01）. Compared with HGgroup, the intracellular fluorescent intensity of DCF increased in IHG group （P〈0.01）, and the activity of T-SOD decreased （P〈0.01）, the level of GSH decreased （P〈0.01l）, the level of GSH increased （P〈0.05）.Conclusion： Intermittent high glucose causes a more significant damage for oxidative stress to GMC.

关 键 词：波动性高糖 肾小球系膜细胞 氧化应激 

分 类 号：R33[医药卫生—人体生理学]