中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2011年
11期
998-1001
,共4页
视网膜%促红细胞生成素%胚胎发育
視網膜%促紅細胞生成素%胚胎髮育
시망막%촉홍세포생성소%배태발육
Retina%Erythropoietin%Embryonic development
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12 d(E12 d)、孕16 d(E16 d)、孕20 d(E20 d)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EPO mRNA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,E12 d、E16 d、E20 d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E16 d最高,E20 d次之,成年组最低.E16 d、E20 d和成年组大鼠视网膜中EPO mRNA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.
揹景 促紅細胞生成素(EPO)錶達于造血組織和神經組織,髮揮促紅細胞生成和神經保護的作用.研究證實,EPO在胚胎髮育的腦組織中有錶達,但其在神經組織來源的視網膜中的錶達情況研究較少.目的 觀察大鼠視網膜胚胎期髮育過程中EPO的錶達變化,探討其與視網膜髮育的關繫.方法 于Wistar母鼠孕12 d(E12 d)、孕16 d(E16 d)、孕20 d(E20 d)時氯胺酮痳醉後剖腹取胎,穫得各階段的胚胎鼠,每組孕鼠各5隻,每隻孕鼠任意選取1隻胎鼠；成年組(12月齡)共選Wistar大鼠5隻.成年鼠及胎鼠用頸椎脫臼法處死後立即摘除眼毬,分離視網膜併製作石蠟切片.用免疫組織化學法、半定量逆轉錄—聚閤酶鏈反應(RT-PCR)檢測大鼠視網膜組織中EPO蛋白及EPO mRNA的錶達.結果胚胎各期視網膜神經上皮層及RPE層均有EPO暘性錶達,免疫暘性染色主要位于細胞質和細胞覈；成年鼠EPO暘性錶達主要集中于RGCs層,E12 d、E16 d、E20 d和成年組大鼠視網膜中EPO的錶達量分彆為105.55±10.35、99.35±8.71、83.27±7.84和30.30±3.80,差異有統計學意義(F=76.13,P＜0.01).凝膠成像半定量分析顯示,EPO基因擴增產物的錶達在E16 d最高,E20 d次之,成年組最低.E16 d、E20 d和成年組大鼠視網膜中EPO mRNA錶達的相對值分彆為0.88±0.10、0.86±0.09和0.26±0.03,胚胎鼠明顯高于成年鼠,差異有統計學意義(F=136.81,P=0.00).結論Wistar大鼠視網膜胚胎髮育過程中EPO的錶達呈現由高到低的趨勢,其錶達規律與視網膜髮育各期的組織學改變相吻閤.這種變化可能與大鼠視網膜胚胎期的髮育密切相關.
배경 촉홍세포생성소(EPO)표체우조혈조직화신경조직,발휘촉홍세포생성화신경보호적작용.연구증실,EPO재배태발육적뇌조직중유표체,단기재신경조직래원적시망막중적표체정황연구교소.목적 관찰대서시망막배태기발육과정중EPO적표체변화,탐토기여시망막발육적관계.방법 우Wistar모서잉12 d(E12 d)、잉16 d(E16 d)、잉20 d(E20 d)시록알동마취후부복취태,획득각계단적배태서,매조잉서각5지,매지잉서임의선취1지태서；성년조(12월령)공선Wistar대서5지.성년서급태서용경추탈구법처사후립즉적제안구,분리시망막병제작석사절편.용면역조직화학법、반정량역전록—취합매련반응(RT-PCR)검측대서시망막조직중EPO단백급EPO mRNA적표체.결과배태각기시망막신경상피층급RPE층균유EPO양성표체,면역양성염색주요위우세포질화세포핵；성년서EPO양성표체주요집중우RGCs층,E12 d、E16 d、E20 d화성년조대서시망막중EPO적표체량분별위105.55±10.35、99.35±8.71、83.27±7.84화30.30±3.80,차이유통계학의의(F=76.13,P＜0.01).응효성상반정량분석현시,EPO기인확증산물적표체재E16 d최고,E20 d차지,성년조최저.E16 d、E20 d화성년조대서시망막중EPO mRNA표체적상대치분별위0.88±0.10、0.86±0.09화0.26±0.03,배태서명현고우성년서,차이유통계학의의(F=136.81,P=0.00).결론Wistar대서시망막배태발육과정중EPO적표체정현유고도저적추세,기표체규률여시망막발육각기적조직학개변상문합.저충변화가능여대서시망막배태기적발육밀절상관.
Background Erythropoietin (EPO) was proved to be express in hematopoietic 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 (E12 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,20d) 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± 7.84and 30.30± 3.80 in E12 d rats,E16 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 E16 d rats,E20 d rats and 12-month-old rats respectively,presenting a elevated value in embryonic rats compared with adult rats ( P =0.00).Gel imaging deletion showed that the A value of EPO amplification products was highest in E16 d rats and lowest in adult rats.Conclusions The expression of EPO appears a high to low fashion during the embryonic development of Wistar rats,which is closely associated with the developing procedure of retina.
