可见光对视网膜色素上皮细胞氧化损伤的影响  

Visible light induce oxidative damage to human retinal pigment epithelial cells

作　　者：傅敏[1] 栾洁[1] 张忠红[1] 

机构地区：[1]东南大学附属中大医院眼科,南京210009

出　　处：《中华眼底病杂志》2011年第5期466-470,共5页Chinese Journal of Ocular Fundus Diseases

基　　金：江苏省卫生厅医学重点学科开放课题（7690000011）

摘　　要：目的观察可见光对体外培养视网膜色素上皮（RPE）细胞内活性氧（ROS）、8-羟基一2L脱氧鸟瞟呤（8-OHdG）和8一羟基鸟嘌呤DNA糖苷酶l（h（）GGl）表达的影响。方法人RPE-19细胞按1：4至1：6传代培养。取第4～6代80％融合细胞用于实验。细胞分为白光、红光、蓝光照射组和对照组，细胞平面光照强度为600Lux。白光，红光分别照射6、12、24、48h；蓝光照射1、3、6、12h；对照组以锡纸包裹避光培养。2’，7＇-二氯荧光素二乙酸酯（DCFHDA）法检测细胞内ROS表达量，免疫细胞化学（ICG）法检测细胞内8—0HdG表达量，蛋白免疫印迹法（Western blot）检测细胞内h0GGl表达情况。结果DCFH—DA法检测结果显示，白光、红光照射组分别照射12、24、48h，蓝光照射组照射1、3、6、12h，细胞内ROS表达量均升高且随时间延长而增加，与对照组细胞内ROS表达量比较，差异均有统计学意义（F白光=11．6jl，F红光=6．706，F蓝光=23．259；P〈O．05）。ICG法检测结果显示，白光、红光照射组分别照射12、24、48h，细胞胞浆8～OHdG表达量增高，照射后12、24h表达量随时间延长而增加，48h有所降低，与对照组细胞胞浆8-OHdG表达量比较，差异均有统计学意义（F白光=16．032，F红光=6．378；P〈0．05）；蓝光照射组照射后1、3、6、12h，细胞胞浆8-OHdG表达量均增高，照射后6、12h细胞胞浆8-OHdG表达量较1、3h细胞胞浆8—OHdG表达量有所降低，但与对照组细胞胞浆8-OHdG表达量比较，差异均有统计学意义（F蓝光=19．484，Pd0．01）。Western blot检测结果显示，白光、红光照射组照射12、24、48h，蓝光照射组照射6、12h，细胞内hOGGl表达量均增高，与对照组细胞内hOGGl表达量比较，差异均有统计学意义（F白光=15．121，F红光=21．041；F蓝光=12．479，P〈o．05）。结论白光、红光、蓝光能以时间依赖性方式诱导RPE细胞ROObjective To observe the effect of visible light （white light, red light, blue light） on the expression of reactive oxygen species （ROS）, 8 OHdG and hOGG1 in cultured human retinal pigment epithelial （RPE） cells. Methods Cultured human RPE-19 cells （4th - 6th generations） were divided into white light, red light, blue light and control group. The illumination was 600 Lux. The cells of experimental groups were exposed to white light or red light for 6, 12, 24 and 48 hours, and exposed to blue light for 1, 3, 6 and 12 hours, while cells of the control group were cultured in foil packaged dishes to avoid light. The levels of ROS expression were detected by 2＇, 7＇- dichlorofluorescin-diacetate （DCFH-DA）, the levels of 8-OHdG protein expression were observed by immunocytochemistry （ICC）, and the levels of hOGG1 were measured by western blot. Results Compared to the control group, the ROS expression in RPE cells were increased in white and red light group after 12, 24 and 48 hours and in blue light group after 1, 3, 6 and 12 hours （Fwhite ligh=11. 611, Fred light=6- 706, Fblue light=23. 259; P〈0.05 ）. Additionally, the ROS expression had a tendency to increase gradually along with exposure time. Compared to the control group, the 8-OHdG expression in RPE cells were increased significantly in both white and red light group after 12, 24 and 48 hours and in blue light group after 1, 3, 6 and 12 hours （Fwhite light = 16. 032, Fred light = 6. 378, Fblue light = 19. 484; P〈0.05）. Additionally, the 8-OHdG expression in white and red light group were increased gradually with exposure time but decreased when exposure time was up to 48 hours, while that in blue light group was increased firstly though it started to decrease when exposure time was up to 6 hours. Compared to the control group, the hOGG1 expression in RPE ceils were increased in white and red light group after 12, 24 and 48 hours and in blue light group after 6 and 12 hours （Fwhite light =15. 121, Fred light = 21, 041, Fblu

关 键 词：色素上皮 眼/细胞学 活性氧 脱氧鸟嘌呤核苷酸类 细胞培养技术 

分 类 号：R774.1[医药卫生—眼科]