【摘要】 目的研究实验性氧化损伤的人视网膜色素上皮(RPE)细胞中凋亡相关基因(bcl-2)表达的变化及促红细胞生成素(EPO)对bcl-2基因表达的影响。方法制备RPE细胞氧化损伤模型,加入40U/ml的重组人促红细胞生成素(rhEPO)共同孵育,采用免疫细胞化学法检测bcl-2蛋白的表达,RT-PCR测定bcl-2mRNA的表达量。结果bcl-2在正常RPE细胞有弱表达,在氧化损伤2、8、24h其表达水平依次降低,氧化损伤组在各时间点与正常组比较差异均有统计学意义(P<0.05)。EPO组bcl-2表达变化规律与氧化损伤组一致,但较氧化损伤组明显增加(P<0.05)。bcl-2mRNA表达量与bcl-2蛋白表达的变化趋势一致。结论bcl-2参与视网膜氧化损伤的机制,EPO促进bcl-2在氧化损伤的RPE细胞中的表达。更多还原

【Abstract】 Objective Retinal pigment epithelium(RPE)situate in the outer layer of retina and is easily injured by oxidation.This research was to investigate the effect of erythropoietin(EPO)on the expression of apoptosis-inhibiting gene,bcl-2,in human RPE cells induced by experimental oxidative injuries.MethodsThe RPE cells strain was take out from-70 ℃ refrigetator and put into water bath of 55 ℃-65 ℃.After resuscitation,the RPE cell was inoculated into culture flask and cultured in 5%CO2 incubator.H2O2 of 600 μmol/L was added into RPE cell culture medium to establish the model of oxidative injuries in human RPE.Meanwhile,EPO of 40 U/ml was also added into the culture medium and continued to culture for 2,8 and 24 hours.The expression of bcl-2 protein was studied by cellular immunochemical technique.The bcl-2 mRNA was studied by RT-PCR.ResultsThe brown particles in cytoplasm of normal cultured RPE cell were displayed,demonstrating the expression of bcl-2.bcl-2 expression at 2,8,24 hours after oxidative injuries was 161.83±4.83,156.32±6.40 and 142.10±5.52 respectively,indicating gradually decrease with time(P<0.05).The changes of bcl-2mRNA paralleled to the changes of its protein.The alterative trend of bcl-2 in EPO treatment groups was in coincidence with oxidative injury groups.ConclusionThe expression of bcl-2 is downregulated in human RPE cells of experimental oxidative injuries.Bcl-2 may have a pivotal role in apoptotic pathway of oxidative-damaged RPE cells.EPO can rescue RPE cells from retinal oxidative injuries through upregulation of bcl-2 expression and may represent an important mechanism for therapeutic method of retinal disease related to RPE cells.更多还原