促红细胞生成素在大鼠视网膜胚胎发育中的表达变化  

Changes of erythropoietin expression during the embryonic development period of rat retina

作　　者：袁春燕[1] 孟旭霞[1] 牛膺筠[1] 

机构地区：[1]青岛大学医学院附属医院眼科,266003

出　　处：《中华实验眼科杂志》2011年第11期998-1001,共4页Chinese Journal Of Experimental Ophthalmology

基　　金：青岛市公共领域科技支撑计划资助项目（09-1-1-18-nsh）

摘　　要：背景促红细胞生成素（EPO）表达于造血组织和神经组织，发挥促红细胞生成和神经保护的作用。研究证实，EPO在胚胎发育的脑组织中有表达，但其在神经组织来源的视网膜中的表达情况研究较少。目的观察大鼠视网膜胚胎期发育过程中EPO的表达变化，探讨其与视网膜发育的关系。方法于Wistar母鼠孕12d（E12d）、孕16d（E16d）、孕20d（E20d）时氯胺酮麻醉后剖腹取胎，获得各阶段的胚胎鼠，每组孕鼠各5只，每只孕鼠任意选取1只胎鼠；成年组（12月龄）共选Wistar大鼠5只。成年鼠及胎鼠用颈椎脱臼法处死后立即摘除眼球，分离视网膜并制作石蜡切片。用免疫组织化学法、半定量逆转录-聚合酶链反应（RT—PCR）检测大鼠视网膜组织中EPO蛋白及EPOmRNA的表达。结果胚胎各期视网膜神经上皮层及RPE层均有EPO阳性表达，免疫阳性染色主要位于细胞质和细胞核；成年鼠EPO阳性表达主要集中于RGCs层，E12d、E16d、E20d和成年组大鼠视网膜中EPO的表达量分别为105．55±10．35、99．35±8．71、83．27±7．84和30．30±3．80，差异有统计学意义（F=76．13，P〈0．01）。凝胶成像半定量分析显示，EPO基因扩增产物的表达在E16d最高，E20d次之，成年组最低。E16d、E20d和成年组大鼠视网膜中EPOmRNA表达的相对值分别为0．88±0．10、0．86±0．09和0．26±0．03，胚胎鼠明显高于成年鼠，差异有统计学意义（F=136．81，P=0．00）。结论Wistar大鼠视网膜胚胎发育过程中EPO的表达呈现由高到低的趋势，其表达规律与视网膜发育各期的组织学改变相吻合。这种变化可能与大鼠视网膜胚胎期的发育密切相关。Background Erythropoietin （EPO） was proved to be express in hematopoietie tissue and nervous system and play the effects of stimulating blood cell production and protecting nervous tissue. Researches showed that EPO is expressed in the embryon brain of animal. However,whether EPO exist in nervous-derived retina and its action on retina with the development is concerned. Objective This research was to investigate the expression of EPO during the embryonic development period of rat retina and explore the role of EPO in retina development process. Methods Clean Wistar rats with pregnancy for 12 days,16 days and 20 days were collected, and the embryonic 12-day rats （El2 d,5 rats） ,embryonic 16-day rats （E16 d,5 rats） and embryonic 20-day rats （ E20 d,5 rats） were obtained by caesarean operation, and 5 12-month W istar rats were used as controls. The rats were sacrificed by cervical dislocation and the retinal sections were prepared in the different-embryo-phase （12 d, 16 d, 20 d） and growth phase. The expression of EPO protein and mRNA in rat retina was detected by immunohistochemistry and reverse transcription-polymerase chain reaction （RT-PCR）,respectively. The feed and use of the animals followed the Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. Results EPO was positively expressed in the cytoplasm and nuclei in the neuroepithelial layer and pigment epithelium of every-embryo-phase rats but only in retinal ganglion cell layer in 12- month-old rats. The gray scale values of EPO expression in retina were 105.55 ± 10.35,99.35± 8.71,83.27 ± 7.84 and 30.30± 3.80 in El2 d rats, El6 d rats, E20 d rats and 12-month-old rats respectively with a statistically significant difference （F = 76. 13, P〈0.01 ）. RT-PCR revealed that the relative values of EPO mRNA expression in retina were 0. 876±0. 10,0. 861±0.09 and 0. 256±0.03 in El6 d rats,E20 d rats and 12-month-old rats respectively,presenting a elevated value in emb

关 键 词：视网膜 促红细胞生成素 胚胎发育 

分 类 号：R774.1[医药卫生—眼科]