缺氧条件下维生素E对视网膜色素上皮细胞生长的影响  

Effect of Vitamin E on the damage of retinal pigment epithelium cells induced by hypoxia

作　　者：马健[1] 袁志兰[1] 帅捷[1] 曹鎏[1] 洪瑾[1] 

机构地区：[1]南京医科大学第一附属医院眼科,江苏省南京市210029

出　　处：《中国组织工程研究与临床康复》2007年第23期4527-4530,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30000207)~~

摘　　要：目的:自行设计细胞模型并观察缺氧条件下维生素E对培养视网膜色素上皮细胞生长的影响。方法:实验于2006-10/2007-03在南京医科大学生理实验室完成。①实验操作:取对数生长期的细胞,以正常条件为对照(常规培养和体积分数为0.95的空气,体积分数为0.05的CO2),建立人视网膜色素上皮细胞的缺氧模型(缺氧培养,体积分数为0.95的N2和体积分数为0.05的CO2混合气体,测氧仪测氧浓度<1%,24h),作用于细胞的维生素E浓度分别为0.1,0.2,0.3mol/L。②实验评估:四甲基偶氮唑盐法检测24h细胞活力(以吸光度A值表示);激光共聚焦显微镜检测细胞内活性氧水平;黄嘌呤氧化酶法测定细胞内超氧化物歧化酶活性。结果:①四甲基偶氮唑盐检测细胞生长活力:缺氧组细胞活力低于对照组(P<0.01),缺氧+维生素E组细胞活力有所提高,与缺氧组相比,差异显著(P<0.05)。随着维生素E浓度的提高,活力逐渐升高。②细胞内活性氧水平:与对照组比较,缺氧组细胞内活性氧的产生明显增多;与缺氧组比较,缺氧+维生素E组抑制视网膜色素上皮细胞内活性氧的产生。③超氧化物歧化酶活力检测:与对照组比较,缺氧组超氧化物歧化酶的活力降低;与缺氧组比较,缺氧+维生素E组超氧化物歧化酶活力升高,差异均具有显著性意义(P<0.05)。结论:缺氧引起视网膜色素上皮细胞氧化应激损伤,维生素E抑制了缺氧情况下视网膜色素上皮细胞的损伤作用,维生素E主要通过提高超氧化物歧化酶活性来发挥抗氧化损伤作用,其抗氧化损伤作用有剂量依赖性。AIM： To investigate the effect of Vitamin E on retinal pigment epithelium （RPE） cells exposed to hypoxia in self-designed cell model. METHODS： The experiment was performed in the Physiological Laboratory of Nanjing Medical University from October 2006 to March 2007.①With the normal condition as control （routine culture in 0.95 volume fraction air and 0.05 volume fraction CO2）, RPE cells at logarithmic phase were exposed to hypoxia （0.95 volume fraction N2 and 0.05 volume fraction CO2, oxygen concentration was 〈 1%） for 24 hours to establish hypoxia models. Different concentrations of Vitamin E （0.1, 0.2, and 0.3 mol/L） were added into the anoxic RPE cells. ②MTT assay was used to evaluate the vital force （indicated by value A）. The intracellular reactive oxygen species （ROS） was detected by confocal microscopy with fluorescent probe CM-H2DCFDA.The activity of superoxide dismutase （SOD） was measured by xanthine oxidase method. RESULTS：①MTT assay results showed that the viability of RPE cells exposed to hypoxia was lower than that of control group （P 〈 0.01 ）, and the cell viability was markedly increased by hypoxia and Vitamin E compared with hypoxia group （P 〈 0.05）. Moreover, the validity was gradually elevated with the increase of Vitamin E concentration. ②Compared with control group, the level of intracellular ROS was markedly increased in hypoxia group, but it was inhibited by Vitamin E and hypoxia.③Compared with control group, there was a decrease in the activity of SOD in RPE cells exposed to hypoxia, but an increase in cells in Vitamin E and hypoxia group compared with hypoxia group （P 〈 0.05）. CONCLUSION： Hypoxia leads to the oxidative stress injury of RPE cells. Vitamin E could inhibit this damage in a dose-dependent manner through enhancing the activity of SOD.

关 键 词：老年性黄斑变性 维生素E 缺氧 活性氧 超氧化物歧化酶 

分 类 号：R773[医药卫生—眼科]