过氧化氢对人视网膜色素上皮细胞ARPE-19中20S蛋白酶体表达的上调作用

　　闫焱1，雷霞2，王玲1，徐国彤2

　　作者单位：1（200025）中国上海市，上海交通大学医学院附属瑞金医院眼科；2（200025）中国上海市，中国科学院上海生命科学研究院/上海交通大学医学院健康科学研究所临床视觉研究组

　　作者简介：闫焱，住院医师，在读博士研究生。
 
　　通讯作者：王玲，博士，主任医师，博士生导师，教授[email protected]；徐国彤，博士，研究员，博士生导师，教授[email protected]

　　稿日期：2008-04-29修回日期：2008-05-29

　　Up-regulation effect of hydrogen peroxide on expression of 20S proteasome in ARPE-19

　　Yan Yan1, Xia Lei2, Ling Wang1, Guo-Tong Xu2
1Department of Ophthalmology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China;2Institute of Health Sciences, Shanghai Institutes of Biological Sciences Chinese Academy of Sciences and Shanghai Jiaotong University School of Medicine, Shanghai 200025, China
Correspondence to: Ling Wang. Department of Ophthalmology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China.

　　[email protected];Guo-Tong Xu.Institute of Health Sciences, Shanghai Institutes of Biological Sciences Chinese Academy of Sciences and Shanghai Jiaotong University School of Medicine, Shanghai 200025, [email protected]

　　Received:2008-04-29Accepted:2008-05-29

　　Abstract

　　AIM: To investigate the effect of hydrogen peroxide on 20S proteasome expression induced by oxidative stress in ARPE-19 cells in vitro.

　　METHODS: ARPE-19 were treated with 0, 100 or 300μmol/L hydrogen peroxide for 4 hours, respectively, then the expression and distribution of 20S proteasome in cells were detected by Western Blot and immunofluorescence staining assay respectively.
 
　　RESULTS: 20S proteasome increased significantly in ARPE-19 when it treated by hydrogen peroxide at 100 and 300μmol/L (100μmol/L hydrogen peroxide vs 0, 300μmol/L hydrogen peroxide , P﹤0.05）, and the up-regulation change was dose-dependent. Intracellular distribution of 20S proteasome in ARPE-19 cultured under normal circumstances was around the nuclear membrane. 20S proteasome expression in ARPE-19 increased after treated by hydrogen peroxide at 100 and 300μmol/L, mainly in the nucleus and around the nuclear membrane. 

　　CONCLUSION: 20S proteasome is up-regulated in ARPE-19 treated by hydrogen peroxide. Ubiquitin-proteasome pathway is induced in ARPE-19 by oxidative stress in vitro.

　　KEYWORDS: retinal pigment epithelial cell; oxidative stress; proteasome; hydrogen peroxide Yan Y,Lei X,Wang L, et al. Up-regulation effect of hydrogen peroxide on expression of 20S proteasome in ARPE-19. Int J Ophthalmol(Guoji  Yanke Zazhi) 2008;8(6):1156-1158

　　摘要

　　目的：探讨过氧化氢诱导的氧化应激对人视网膜色素上皮细胞ARPE-19中20S蛋白酶体表达的影响。

　　方法：分别选择0，100，300μmol/L过氧化氢作用ARPE-19细胞4h后收集样本，Western blot方法检测细胞中20S蛋白酶体表达水平，细胞免疫荧光方法检测20S蛋白酶体在细胞中的分布。

　　结果： 100和300μmol/L过氧化氢溶液处理的细胞中20S蛋白酶体的表达和对照组相比明显增加（100μmol/L过氧化氢处理组vs0和300μmol/L过氧化氢处理组，P<0.05），并呈现剂量依赖性。正常培养情况下，细胞内20S蛋白酶体在核周微量表达，100和300μmol/L过氧化氢溶液处理的细胞内20S蛋白酶体和对照组相比明显增加，其分布集中在核内及核膜周围。

　　结论：过氧化氢对ARPE-19的20S蛋白酶体表达有上调作用。

　　关键词：过氧过氢；视网膜色素上皮细胞；氧化应激；蛋白酶体

　　闫焱，雷霞，王玲，等.过氧化氢对人视网膜色素上皮细胞ARPE-19中20S蛋白酶体表达的上调作用.国际眼科杂志2008;8(6):1156-1158