上海交通大学学报（医学版）

›› 2012, Vol. 32 ›› Issue (2): 137-.doi: 10.3969/j.issn.1674-8115.2012.02.003

• 论著（基础研究） • 上一篇    下一篇

促红细胞生成素及其受体在大鼠视网膜光损伤后的表达

宫媛媛, 童念庭, 顾 青, 吴星伟, 孙晓东   

  1. 上海交通大学附属第一人民医院眼科, 上海 200080
  • 出版日期:2012-02-28 发布日期:2012-02-28
  • 通讯作者: 孙晓东, 电子信箱: xdsun@sjtu.edu.cn。
  • 作者简介:宫媛媛（1973—）, 女, 副主任医师, 博士；电子信箱: gyydr@126.com。
  • 基金资助:

    国家重点基础研究发展计划（“九七三”计划）子课题（2011CB707506）；上海市眼底病重点实验室开放课题基金（07Z22911）

Expression of erythropoietin and erythropoietin receptor after light-induced retinal injury in rats

GONG Yuan-yuan, TONG Nian-ting, GU Qing, WU XING-wei, SUN Xiao-dong   

  1. Department of Ophthalmology, the First People's Hospital, Shanghai Jiaotong University, Shanghai 200080, China
  • Online:2012-02-28 Published:2012-02-28
  • Supported by:

    National Key Basic Research Development Program of China, 973 Program, 2011CB707506；Shanghai Key Laboratory for Ocular Fundus Diseases Foundation, 07Z22911

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摘要：

目的 观察大鼠视网膜光损伤后促红细胞生成素（EPO）及其受体（EPOR）的表达变化。方法 选用32只雄性SD大鼠，其中29只建立视网膜光损伤大鼠模型（光损伤组），另3只作为正常对照组。取正常对照组大鼠视网膜组织以及光损伤组大鼠光损伤后0、6、12、24、48、72 h和7、14 d的视网膜组织，分别采用Western blotting和RT-PCR法检测EPO、EPOR蛋白和mRNA的表达，并采用免疫组织化学法定位观察EPO和EPOR蛋白表达。结果 Western blotting检测显示：视网膜光损伤后，EPO和EPOR蛋白表达增加，光损伤后48 h达到高峰；光损伤组EPO蛋白的相对表达量在光损伤后12、24、48、72 h显著高于正常对照组（P<0.05或P<0.01），光损伤组EPOR蛋白的相对表达量在光损伤后6、12、24、48、72 h和7、14 d显著高于正常对照组（P<0.05或P<0.01）。RT-PCR检测显示：视网膜光损伤后，EPO和EPOR mRNA表达增加，分别于光损伤后24 h和48 h达到高峰；光损伤组EPO mRNA的相对表达量在光损伤后12、24、48、72 h和7、14 d显著高于正常对照组（P<0.05或P<0.01），光损伤组EPOR mRNA的相对表达量在光损伤后6、12、24、48、72 h和7、14 d显著高于正常对照组（P<0.05或P<0.01）。免疫组织化学定位观察显示：视网膜光损伤后24 h，光感受器内外段EPOR蛋白表达略增强，EPO蛋白表达相对较弱。结论 大鼠视网膜光损伤后EPO和EPOR蛋白及mRNA表达增加，有一定的时效性，提示内源性EPO和EPOR参与视网膜光损伤的修复机制。

关键词: 促红细胞生成素, 受体, 视网膜, 光损伤

Abstract:

Objective To investigate the expression of erythropoietin (EPO) and erythropoietin receptor (EPOR) after light-induced retinal injury in rats. Methods Thirty-two male SD rats were selected, light-induced retinal injury models were established in 29 rats (light-induced retinal injury group), and the other 3 rats were served as normal control group. The retinal tissues of rats in normal control group and those of rats in light-induced retinal injury group at the time points of 0 h, 6 h, 12 h, 24 h, 48 h, 72 h, 7 d and 14 d after light-induced retinal injury were obtained, the expression of EPO and EPOR protein and mRNA was detected by Western blotting and RT-PCR respectively, and the location of expression of EPO and EPOR protein was determined with immunohistochemical method. Results Western blotting detection revealed that the expression of EPO and EPOR protein increased after light-induced retinal injury, and reached the peak 48 h after light-induced retinal injury. The relative expression of EPO protein in light-induced retinal injury group was significantly higher than that in normal control group at the time points of 12 h, 24 h, 48 h and 72 h after light-induced retinal injury (P<0.05 or P<0.01), and the relative expression of EPOR protein in light-induced retinal injury group was significantly higher than that in normal control group at the time points of 6 h, 12 h, 24 h, 48 h, 72 h, 7 d and 14 d after light-induced retinal injury (P<0.05 or P<0.01). RT-PCR detection demonstrated that the expression of EPO and EPOR mRNA increased after light-induced retinal injury, and reached the peak 24 h and 48 h after light-induced retinal injury respectively. The relative expression of EPO mRNA in light-induced retinal injury group was significantly higher than that in normal control group at the time points of 12 h, 24 h, 48 h, 72 h, 7 d and 14 d after light-induced retinal injury (P<0.05 or P<0.01), and the relative expression of EPOR mRNA in light-induced retinal injury group was significantly higher than that in normal control group at the time points of 6 h, 12 h, 24 h, 48 h, 72 h, 7 d and 14 d after light-induced retinal injury (P<0.05 or P<0.01). Immunohistochemical observation indicated that 24 h after light-induced retinal injury, the expression of EPOR protein was slightly enhanced and that of EPO protein was relatively lower in the outer and inner segment of photoreceptor. Conclusion The expression of EPO and EPOR protein and mRNA in retinal tissues may time-dependently increase after light-induced retinal injury in rats, which indicates that endogenous EPO and EPOR may participate in the restoration mechanism of light-induced retinal injury.

Key words: erythropoietin, receptor, retina, light-induced injury