[1]俞永珍,徐哲,邹秀兰,等. 蓝光诱导氧化应激反应参与视网膜色素上皮细胞凋亡机制研究[J].眼科新进展,2015,35(6):520-524.[doi:10.13389/j.cnki.rao.2015.0141]
　YU Yong-Zhen,XU Zhe,ZOU Xiu-Lan,et al. Role of oxidative stress induced by blue light in human retinal pigment epithelium cells apoptosis[J].Recent Advances in Ophthalmology,2015,35(6):520-524.[doi:10.13389/j.cnki.rao.2015.0141]
点击复制

 蓝光诱导氧化应激反应参与视网膜色素上皮细胞凋亡机制研究
分享到：

《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
35卷
期数:
2015年6期
页码:
520-524
栏目:
实验研究
出版日期:
2015-06-05

文章信息/Info

Title:
 Role of oxidative stress induced by blue light in human retinal pigment epithelium cells apoptosis
作者:
 俞永珍; 徐哲; 邹秀兰; 张楚; 王观峰; 邹玉平
 510010　广东省广州市，广州军区广州总医院眼科
Author(s):
 YU Yong-Zhen;  XU Zhe;  ZOU Xiu-Lan;  ZHANG Chu;  WANG Guan-Feng ; ZOU Yu-Ping
 Department of Ophthalmology, Guangzhou Ceneral Hospital of Guangzhou Military Command, Guangzhou 510010 , Guangdong Province , China
关键词:
 ; 凋亡; 活性氧; 氧化应激; 人视网膜色素上皮细胞
Keywords:
 ; apoptosis ;  reactive oxygen species ;  oxidative stress;  human retinal pigment epithelium cell
DOI:
10.13389/j.cnki.rao.2015.0141
文献标志码:
A
摘要:
 目的　研究氧化应激反应参与蓝光诱导体外培养的人视网膜色素上皮细胞（ｈｕ-ｍａｎｒｅｔｉｎａｌｐｉｇｍｅｎｔｅｐｉｔｈｅｌｉｕｍ，ｈＲＰＥ）凋亡的作用机制及其与时间的关系。方法　用ＬＥＤ蓝光光源建立蓝光损伤体外培养的ｈＲＰＥ细胞模型，蓝光辐射强度为（４．０±０．５）ｍＷ?ｃｍ－２，并将细胞分为光照时间０ｈ（正常对照组）、０．５ｈ、１ｈ、２ｈ、３ｈ、４ｈ、５ｈ、６ｈ、１２ｈ组。透射电镜观察细胞的超微结构，流式细胞术检测细胞凋亡率及细胞内活性氧水平，ＷｅｓｔｅｒｎＢｌｏｔ技术检测Ｂａｘ、Ｂｃｌ-２、Ｃａｓｐａｓｅ-３蛋白的表达情况。结果　各光照组细胞均有不同程度的胞浆减少、空泡状改变、线粒体肿胀、微绒毛减少或脱落。正常对照组、光照１ｈ、２ｈ、３ｈ、４ｈ、５ｈ、６ｈ、１２ｈ的细胞凋亡率分别为（５．３０±０.６４）％、（５．９０±０．８９）％、（６．８０±０.９８）％、（７．７０±０．７６）％、（９．６０±１．１０）％、（１１．４５±２.６０）％、（１４．９０±１．６０）％及（２３.９０±１．２０）％，与正常对照组比较，光照４ｈ后细胞凋亡率明显升高，差异均有统计学意义（均为Ｐ＜０．０５）。正常对照组、０．５ｈ、１ｈ、２ｈ、３ｈ、４ｈ、５ｈ、６ｈ组ｈＲＰＥ细胞生成的ＲＯＳ相对量分别为（１２．９０±１．１８）％、（２０．０９±０．６３）％ 、（２３．９１±１．４７）％、（２９．１４±１．９４）％、（３４．３０±１．８４）％、（３８．９７±１.６８）％、（４４．０８±０．８３）％、（５７．７６±２．８０）％，光照０．５ｈ后细胞产生的ＲＯＳ明显升高，与正常对照组比较，差异均有统计学意义（均为Ｐ＜０．０５）。Ｃａｓｐａｓｅ-３及Ｂａｘ在３ｈ后轻微上调，而５～６ｈ后表达上调较明显，Ｂｃｌ-２的表达则在３ｈ后下调明显，与正常对照组比较，差异均有统计学意义（均为Ｐ＜０．０５）。结论　蓝光可通过产生的氧化应激反应激活细胞凋亡系统导致细胞损伤，光照持续３ｈ时细胞可能出现了损伤但不明显，光照５～６ｈ后出现了较为明显的细胞凋亡。
Abstract:
 Objective To research the mechanism of oxidative stress mediating the cultured human retinal pigment epithelium cell ( hRPE) apoptosis induced by blue light and the relationship with time. Methods The blue light damage model of cultured hRPE cells in vitro with LED blue light density of (4. 0 +0. 5) mW . cm -2 adjusted by FL-ID blue light illumination meter was established, and the illununation time was sat as o hour ,0. 5 hour . I hour ,2 hours ,3 hours ,4 hours ,5 hours ,6 hours , 12 hours , then the cells are grouped according to illumination time. Cell ultrastructures were observed by transnussion electron microscope , the rates of total cell apoptosis and the levels of reactive oxygen species( ROS) were measured with flow cytometry , and the expressions of Bax,Bcl-2 and Caspase-3 were detected by Western Blot. Results The cells and michondria became vacuolar or swollen, cytoplasm and microvilli were dinunished in each illununation group. The total apoptosis rates at o hour, I hour ,2 hours , 3 hours ,4 hours .5 hours.6 hours. 12 hours were ( 5. 30 + 0. 64 ) % , (5. 90 + 0. 89 ) % . ( 6. 80 + 0. 98 ) % , ( 7. 70 + 0. 76 ) % . ( 9. 60 + 1. 10) % , ( 11. 45 + 2. 60 ) % . ( 14. 90 + 1. 60 ) % and (23. 90 + 1. 20 )% , respectively. The apoptosis rate increased after illumination for 4 hours compared with control group , there were significant differences ( all P < 0. 05 ) . The levels of ROS at o hour .0. 5 hour . I hour .2 hours .3 hours ,4 hours .5 hours .6 hours were ( 12. 90 + 1. 18) % . ( 20. 09 +0. 63 ) % . ( 23. 91 + 1. 47 ) % . (29. 14 + 1. 94) % , ( 34. 3 + 1. 84 ) % . ( 38. 97 + 1. 68 ) % . ( 44. 08 + 0. 83 ) % , ( 57. 76 + 2. 80 ) % , respectively , the levels of ROS increased sigruficantly after illumination for 0. 5 hour compared with control group ( all P < 0. 05 ) . The expressions of Caspase-3 and Bax were up-regulated slightly after illumination for 3 hours and up-regulated significantly after illununation for 5 hours t0 6 hours.while the expression of Bcl-2 after cell was down-regulated after illununated for 3 hours. there were significant differences compared with control group ( all P < 0. 05 ) . Conclusion Blue light can cause cell apoptosis in hRPE probably motivated by oxidative stress, the apoptosis phenomenon is not obvious after exposing for 3 hours , but obvious after exposing for 5 hours t0 6 hours.

相似文献/References:

[1]李晓艳 张卯年.褪黑素对糖尿病大鼠早期视网膜神经细胞凋亡的影响[J].眼科新进展,2012,32(6):000.
[2]钟晓东 罗卫民 刘越峰.白藜芦醇对糖尿病性白内障大鼠晶状体上皮细胞凋亡及 bcl-2和 bax表达的影响[J].眼科新进展,2012,32(7):000.
[3]周海燕 高晨 周丽雅 张景科 王小梅 田蔓男.依达拉奉对大鼠视神经钳夹伤后视网膜细胞的保护作用[J].眼科新进展,2012,32(12):000.
[4]杜安杰 任兵 高晓唯 付燕 赵旭东 郭继华.氨基胍对氧诱导的视网膜病变小鼠视网膜神经细胞的保护作用[J].眼科新进展,2012,32(12):000.
[5]梁静 万新顺.川芎嗪对慢性高眼压大鼠视网膜神经节细胞凋亡的影响[J].眼科新进展,2013,33(8):000.
[6]李艳 潘斌 李筱荣 袁佳琴 张杰.糖尿病大鼠视网膜PEDF表达与神经退行性改变[J].眼科新进展,2012,32(8):000.
[7]刘建伟 赵桂秋 杜兆东 张丽娜 张振华.大鼠眼钝挫伤后晶状体上皮细胞的凋亡及Survivin基因对凋亡的影响[J].眼科新进展,2012,32(11):000.
[8]张千帆 刘瑞敏 穆红梅 郭洋 申飞 刘莹.白藜芦醇对视网膜母细胞瘤Y79细胞增殖的影响及其机制[J].眼科新进展,2012,32(11):000.
[9]付馨余 邹文进 黄明汉 赵静博.多西环素对碱烧伤大鼠角膜组织中NF-κB和bcl-2表达的影响[J].眼科新进展,2013,33(4):000.
[10]陈莲 石晶明.青光眼患者视网膜神经节细胞凋亡机制的研究进展[J].眼科新进展,2013,33(4):000.
[11]董凯 柯根杰 武立云 朱子诚 许迅 孙晓东. 视网膜脱离后光感受器细胞死亡规律的实验研究[J].眼科新进展,2014,34(2):144.
[12]周俊,何湘珍,肖启国. 金樱子通过增加Bcl-2/Bax表达比抑制大鼠糖尿病性白内障晶状体上皮细胞的凋亡[J].眼科新进展,2014,34(4):314.[doi:10.13389/j.cnki.rao.2014.0085]
[13]冯冬梅,朱鸿,施彩虹. CXCR3、IP10对高糖诱导的人脐静脉内皮细胞凋亡的影响[J].眼科新进展,2014,34(7):628.[doi:10.13389/j.cnki.rao.2014.0172]
[14]马岚,张奕霞,练海东,等. 右美沙芬对糖尿病视网膜神经病变中Caspase-3表达的影响[J].眼科新进展,2015,35(5):432.[doi:10.13389/j.cnki.rao.2015.0117]
　MA Lan,ZHANG Yi-Xia,LIAN Hai-Dong,et al. Effects of dextromethorphan on expression of Caspase-3 in retinal nerve cell of rats with diabetic retinopath[J].Recent Advances in Ophthalmology,2015,35(6):432.[doi:10.13389/j.cnki.rao.2015.0117]

备注/Memo

备注/Memo:
 广东省科技计划项目（编号：２０１１Ｂ０３１８００２０２）；广东省科学自然基金项目（编号：Ｓ２０１３０１００１２０４５）
更新日期/Last Update: 2015-06-01