间歇性高糖对胰岛微血管内皮细胞凋亡和氧化应激损伤的影响  

Effects of intermittent high concentration of glucose on rat islet microvascular endothelial cells apoptosis and damage of oxidative stress

作　　者：孙嘉[1] 陈新[1] 陈宏[1] 蔡德鸿[1] 

机构地区：[1]南方医科大学珠江医院内分泌科,广州510282

出　　处：《中华损伤与修复杂志（电子版）》2012年第3期24-27,共4页Chinese Journal of Injury Repair and Wound Healing(Electronic Edition)

基　　金：广东省医学科学技术研究基金项目(B2011207)

摘　　要：目的探讨间歇性高糖对胰岛微血管内皮细胞凋亡和氧化应激损伤的影响。方法以体外不同葡萄糖浓度DMEM培养基培养的大鼠胰岛微血管内皮细胞为研究对象,分为正常葡萄糖组、恒定性高糖组和间歇性高糖组,连续培养7d。采用DNA片段化检测及流式细胞仪测定胰岛微血管内皮细胞的凋亡,连续光谱荧光仪检测细胞凋亡蛋白酶Caspase-3活性。通过测定细胞活性氧族水平、线粒体膜电位及黄嘌呤氧化酶评估不同培养条件下对胰岛微血管内皮细胞氧化应激的影响。结果间歇性高糖组较正常葡萄糖组和恒定性高糖组的细胞凋亡明显增多,差异具有统计学意义(P<0.001);间歇性高糖组细胞内活性氧簇、黄嘌呤氧化酶水平显著高于恒定性高糖组和正常葡萄糖组差异具有统计学意义(P<0.001)。间歇性高糖组线粒体膜电位水平显著低于恒定性高糖组和正常葡萄糖组差异具有统计学意义(P<0.001)。结论间歇性高糖较恒定性高糖能够促进胰岛微血管内皮细胞氧化应激损伤及增加细胞凋亡。Objective To investigate the effects of intermittent high concentration of glucose on rat islet microvascular endothelial cells （IMECs） apoptosis and damage of oxidative stress. Methods The rat IMECs cultured in vitro, as experiment subjects, were randomly divided into three groups, i.e. （1）normal concentration of glucose group （NG, n=9）, （2）constant high concentration of glucose group （CHG, n=9）, and （3）intermittent high concentration of glucose group （IHG, n=9）. The IMECs of each group were cultured for 7 days respectively. The apoptosis of IMECs was measured by DNA fragmentation and flow cytometry, while the activity of apoptotic protease Caspase-3 was detected by continuous spectrum fluorescence spectrometer to find apoptosis pathway. The levels of reactive oxygen species （ROS）, mitochondrial membrane potential （MMP）, and intracellular xanthine oxidase （XOD） were used to evaluate the oxidative stress of IMECs in three different groups. Results There was a significantly higher apoptosis of IMECs in IHG group than in NG group and CHG group （P0.001）, while the levels of ROS and XOD in IHG group were much higher than in NG group and CHG group （P0.001）. However, there was a significant reduction of MMP in IHG group than in CHG group （P0.001）. Conclusions Intermittent high concentration of glucose can induce much more severe apoptosis and damage of oxidative stress of IMECs in vitro than constant high concentration of glucose.

关 键 词：胰岛微血管内皮细胞 间歇性高糖 氧化应激 凋亡 

分 类 号：R587.1[医药卫生—内分泌]