雌激素对蓝光及过氧化氢诱导的ARPE-19细胞内活性氧生成的影响

　　雷少波，唐罗生，朱晓华，段宣初
 
　　作者单位：（410011）中国湖南省长沙市，中南大学湘雅二医院眼科

　　作者简介：雷少波，男，博士研究生，主要从事眼底病临床、科研、教学工作。
 
　　通迅作者：唐罗生，男，教授，博士生导师，主要从事视网膜病和白内障的临床和基础研究.tangls57@ vip.sina.com

　　收稿日期：2008-04-07修回日期：2008-06-01

　　Effects of estrogen on ROS generation induced by blue light and hydrogen peroxide in human retinal epithelial cell ARPE-19
Shao-Bo Lei,Luo-Sheng Tang,Xiao-Hua Zhu,Xuan-Chu Duan
Department of Ophthalmology, the Second Xiangya Hospital, Central South University, Changsha 410017, Hunan Province, China

　　Correspondence to:Luo-Sheng Tang. Department of Ophthalmo-logy, the Second Xiangya Hospital, Central South University, Changsha 410017, Hunan Province,[email protected]

　　Received:2008-04-07Accepted:2008-06-01

　　Abstract

　　AIM: To determine whether estrogen suppresses intracellular ROS generation induced by blue light illumination and hydrogen peroxide in human retinal epithelial cell ARPE-19.

　　METHODS: Cultured human retinal epithelial cell line ARPE-19 was pretreated with 17β-estradiol (at concen-tration of 10nmol/L, 100nmol/L, 1μmol/L ) for 2 hours, before given blue light illumination（470±20nm，2 000±500lx）or 200μmol/L H2O2 treatment for a variety of time. Intracellular ROS generation was detected by DCFH-DA fluorescent probe and flow cytometry before oxidant treatment and 30 minutes, 1, 2, 4 hours afterward.

　　RESULTS: DCFH-DA fluorescence intensity was significantly increased after blue light illumination or H2O2 treatment, indicating that ROS generation was success-fully induced in ARPE-19 cell, the increase in DCFH-DA fluorescence intensity was significantly suppressed by 10nmol/L, 100nmol/L, 1μmol/L 17β-E2 in a dose-dependent manner.

　　CONCLUSION: 17β-E2 attenuates blue light and H2O2- induced ROS generation in ARPE-19 cell

　　KEYWORDS: estrogen; retinal pigment epithelium; oxidative stress; reactive oxygen species; phototoxicity
Lei SB, Tang LS, Zhu XH, et al. Effects of estrogen on ROS generation induced by blue light and hydrogen peroxide in human retinal epithelial cell ARPE-19. Int J Ophthalmol(Guoji Yanke Zazhi)2008;8(6):1133-1136

　　摘要

　　目的：观察17-β雌二醇（E2）对蓝光及过氧化氢（H2O2）诱导的人视网膜色素上皮ARPE-19细胞内活性氧（reactive oxygen species, ROS）生成的影响

　　方法：对培养的ARPE-19细胞予以浓度为10nmol/L, 100nmol/L,1μmol/L的E2预处理2h后接受蓝光照射（470±20nm，2000±500lx）或200μmol/L H2O2处理，在处理前、处理后30min；1，2，4h行2′, 7′－二氢二氯荧光黄双乙酸钠（DCFH-DA）细胞内ROS荧光染色，以490nmol/L荧光显微镜观察照相，收集细胞后以流式细胞仪检测荧光强度。

　　结果：蓝光照射和过氧化氢均可显著增强ARPE-19细胞内的ROS荧光，提示蓝光照射和过氧化氢处理均成功诱导氧化应激，10nmol/L、100nmol/L、1μmol/L 17β-雌二醇剂量依赖性地减弱蓝光照射和过氧化氢处理后ARPE-19细胞内的ROS荧光增强的幅度。

　　结论：蓝光照射及过氧化氢处理可诱导ARPE-19细胞内活性氧水平增高；17β－雌二醇可剂量依赖性地抑制蓝光照射诱导的ARPE-19细胞内活性氧水平增高。
 
　　关键词：雌激素；视网膜色素上皮细胞；氧化应激；活性氧；光毒性

　　雷少波，唐罗生，朱晓华，等.雌激素对蓝光及过氧化氢诱导的ARPE-19细胞内活性氧生成的影响.国际眼科杂志2008;8(6):1133-1136