不同来源糖化终末产物对INS-1细胞氧化应激的影响张月成林宁秦利.DOCVIP

基金项目：国家自然科学基金；上海交通大学医学院院校基金（2008XJ020）。 作者单位：200092，上海交通大学医学院附属新华医院内分泌科。第一作者现在江苏大学附属昆山市第一人民医院 通讯作者：秦利，电子信箱：qinli@ 不同来源糖化终末产物对INS-1细胞氧化应激的影响 张月成 林宁 秦利 苏青 摘 要： 目的：研究不同来源糖化终末产物(AGEs)对INS-1细胞氧化应激的影响。方法：制备不同浓度AGE-1、AGE-2和AGE-3，干预INS-1细胞一定时间后，MTT比色法检测细胞活力变化。运用活性氧(ROS)捕获剂双氢-乙酰乙酸二氯荧光黄(DCFH-DA)孵育细胞，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。结果：AGE-2和AGE-3处理组INS-1细胞活力降低(P0.05)，细胞活力与AGE-2和AGE-3浓度呈负相关(r=-0.871/-0.912，P0.05),胞内ROS水平升高(P0.05)，但AGE-1处理组差异无显著性(P0.05)。AGE-2和AGE-3对INS-1细胞活力和胞内ROS水平的影响具有浓度依赖性和时间依赖性：浓度越高、时间越长INS-1细胞活力的降低和胞内ROS水平升高越显著。结论：AGE-2和AGE-3可诱导INS-1细胞氧化应激和细胞损伤。 关键词：糖化终末产物；胰岛素?细胞；氧化应激；糖尿病 Effects of different source of advanced glycation end products on oxidative stress in INS-1 cells Abstract: Objective: To explore the effects of different source of advanced glycation end products(AGEs) on the oxidative stress in INS-1 cells. Methods: After being cultured with AGEs for some time, cell viability was detected by MTT; 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. Results: Viability of INS-1 was inhibited by AGE-2 and AGE-3 in a dose and time-dependent manner (P0.05). The cell viability was negatively correlated with dose of AGE-2 and AGE-3 (r=-0.871/-0.912，P0.05). Intracellular fluorescent intensity of DCF was markedly elevated in a dose and time-dependent manner in the AGE-2 and AGE-3 treated groups than the AGE-1 treated group. Conclusion: AGE-2 and AGE-3 could induce oxidative stress and cell damage in INS-1 cells. Key words： advanced glycation end products; ? cells; oxidative stress; diabetes 糖化终末产物(advanced glycosylation end-products, AGEs)是葡萄糖等碳水化合物和含氨基生物分子（主要为蛋白质，也包括核酸和脂类）通过非酶糖化（non-enzymatic glycation）缓慢作用而形成的一类结构复杂的化合物[1]。虽然早年人们是通过研究葡萄糖和蛋白质的非酶糖化作用而认识AGEs的，但现已清楚除葡萄糖外其它碳水化合物也能与蛋白质形成AGEs。Takeuchi等根据形成AGEs的碳水化合