过氧化氢诱导视网膜色素上皮细胞氧化损伤模型的建立  

Development of a model of oxidative damage of human retinal pigment epithelial cells induced by hydrogen peroxide

作　　者：杨影[1] 曲超[1] YANG Ying QU Chao(Department of Ophthalmology, Sichuan Academy of Medi- cal Sciences ＆ Sichuan Provincial People＇s Hospital, Chengdu ,610072, China)

机构地区：[1]四川省医学科学院.四川省人民医院眼科,四川成都610072

出　　处：《实用医院临床杂志》2017年第4期18-21,共4页Practical Journal of Clinical Medicine

基　　金：四川省科技厅应用基础研究项目(编号:30504010256)

摘　　要：目的探讨视网膜色素上皮细胞氧化损伤模型的构建方法。方法将培养的人视网膜色素上皮细胞分为对照组和实验组,实验组加入50、100、200、300、400μmol/LH_2O_2,对照组不加入H_2O_2,采用MTT法检测细胞活力,在培养箱内分别培养2、8、24 h,采用CCK-8检测细胞抑制率,用比色法观察每组中超氧化物歧化酶(SOD)及丙二醛(MDA)含量的变化,流式细胞术检测细胞凋亡。结果人RPE细胞经200μmol/LH_2O_2处理2、8、24 h后,与对照组比较细胞的活力、抗氧化物酶SOD活性均出现显著降低(P<0.05),MDA含量均有显著升高(P<0.01),细胞凋亡率较对照组显著升高,差异有统计学意义(P<0.01)。结论 200μmol/LH_2O_2作用处理人视网膜色素上皮细胞是体外模拟视网膜色素上皮细胞氧化损伤的良好模型。Objective To develop a model of oxidative damage of retinal pigment epithelial （RPE） cells. Methods Cultured human RPE cells were divided into normal group and experimental group. The experimental group was treated with 50,100,200,300 and 400 μmol/L H_2O_2while the control group was not treated with H_2O_2. The cell growth inhibition rate was detected with MTT assay, The incubator was cultured for 2,8, and 24 h and the cell inhibition rate was detected by using CCK-8. The changes in superoxide dis- mutase （SOD） and malondialdehyde （MDA） were measured with colorimetry method. Flow cytometry was used to detect the cell apop- tosis rate. Results Compared to the control group, the vitality of humna RPE cells and the activity of antioxidant enzyme SOD were sig- nificantly reduced after 2,8 and 24 hours of treatment （P 〈 0. 05 ）, but MDA content was significantly increased （P 〈 0. 0l ）. Apopto- sis rate were significantly higher in the control group when compared to the experimental group .（ P 〈 0. 01 ）. Conclusion Human RPE ceils treated with 200 μomol/L H_2O_2is an appropriate model of oxidative damage of RPE cell in vitro.

关 键 词：过氧化氢 视网膜色素上皮细胞 氧化损伤 

分 类 号：R774.1[医药卫生—眼科]