缺氧诱导体外培养人视网膜色素上皮细胞凋亡

目的:了解缺氧条件下体外培养人视网膜色素上皮(retinal pigment epithelium,RPE)细胞的凋亡情况。方法:将培养的人RPE细胞置于含10mL/LO2、50mL/LCO2和940mL/LN2的培养箱内建立缺氧模型。于缺氧后1,3,6,12,24h,利用扫描电镜、透射电镜、脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(terminal-deoxynucle-otidyl transferase mediated nick end labeling,TUNEL)技术和流式细胞仪测定RPE细胞凋亡水平。结果:常氧状态下RPE细胞生长良好,几乎没有凋亡(TUNEL法测凋亡指数为1.2)。缺氧条件下,RPE细胞发生了不同程度的凋亡,缺氧后3h凋亡水平达峰值(凋亡指数为34.43)。结论:缺氧可导致体外培养的人RPE细胞凋亡,提示RPE凋亡可能参与了某些缺血缺氧性眼病的发生。

Hypoxia induces apoptosis of the cultured human retinal pigment epithelial cells

AIM:To evaluate the effect of hypoxia on apoptosis of cultured human retinal pigment epithelial(RPE)cells.METHODS:In the hypoxia model,human REP cells were cultured in a special incubator containing volume fraction of 10mL/L O2,50mL/L CO2 and 940mL/L N2 for 0 hour,1 hour,3,6,12 and 24 hours.The level of apoptosis was individually measured with scanning and transmis-sion electron microscopy,terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)and flow cytometry.RESULTS:RPE cells grew well under normoxic conditions.The apoptosis was significantly activated and peaked at 3 hours in the cultured RPE cells under hypoxia.CONCLUSION:Hypoxia can induce apoptosis in the cultured RPE cells,suggesting that RPE apoptosis might play a role in ischemic ocular diseases.