医学论文-波动性高糖对大鼠肾小球系膜细胞氧化应激的影响.docVIP

医学论文-波动性高糖对大鼠肾小球系膜细胞氧化应激的影响 ????????????????? 作者：田晓玲，郑景晨，章海凌，沈飞霞，刘芳 【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(5.5 mmol/L，NG)、稳定高糖组(25 mmol/L,HG)、波动性高糖组（5.5 mmol/L或25 mmol/L，每24 h交替，IHG），培养GMC 6 d，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。 【关键词】? 波动性高糖；肾小球系膜细胞；氧化应激 ??? Abstract:? 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（5.5 mmol/l alternating with 25 mmol/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 ma1ondialdehyde（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 HG group，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＜0.01），the level of GSH increased（P＜0.05）.Conclusion：Intermittent high glucose causes a more signific