【摘要】 目的研究促红细胞生成素(EPO)及其受体(EPOR)在人视网膜色素上皮(RPE)细胞化学低氧损伤中的表达及意义。方法原代培养人胚胎RPE细胞,应用不同浓度氯化钴造成RPE细胞化学缺氧,应用四甲基偶氮唑蓝(MTT)比色法检测RPE细胞增生活力变化,采用免疫组织化学法检测EPO和EPOR在RPE细胞中的表达,应用W esternb lot法检测EPO和EPOR蛋白的表达变化。结果不同浓度的CoC l2均促进细胞生长。正常人胚胎RPE细胞检测到EPO和EPOR的弱表达,CoC l2处理组检测到EPO和EPOR的强表达。结论化学缺氧促进人胚胎RPE细胞的增生,EPO及其受体EPOR可能在这个过程中起重要作用。更多还原

【Abstract】 ObjectiveRecent studies demonstrated that erythropoietin（EPO） is a multifunctional biological reactive material.This study aimed to assess the expression of EPO and erythropoietin receptor（EPOR） in human retinal pigment epithelial（RPE）cells exposed to Cobalt Chloride(CoCl₂) in vitro.MethodsCultured human fetal RPE cells were exposed to different concentration（50,100,150,200,300μmol/L） of CoCl₂in vitro.The 3-（4,（5-dimethylthiazole-2y1））-2,5-diphenyl tetrazolium bromide（MTT） cell viability assay was used to assess the effects of hypoxic injury caused by CoCl₂on primary human fetal RPE cell cultures.Immunocytochemical staining and Western blot were used to determine the expression of EPO and its receptor EPOR in human fetal RPE cells.ResultsImmunohistochemical staining showed specific brown yellow staining of EPO and EPOR at cytoplasm and membrane in human fetal RPE cells.The quantity of EPO positive cells was 102.2±3.0 and that of EPOR was 124.2±1.7 in normal RPE cells and was gradually increased after exposure to CoCl₂ with the most cell number at（200μmol/L） group（422.75±10.31,457.75±7.18 respectively）.The A value was（0.0882±0.0010） in the control group and the A value was enhanced with the increased concentration of CoCl₂ exposure and reached the peak value（（0.1167±0.0037）） in the（200μmol/L） group and began to decrease in the concentration of 300μmol/L CoCl₂,but was still higher than that of the normal group,showing that hypoxia caused by CoCl₂ promoted viability and proliferation of RPE cells.The protain expression of EPO and EPOR was 0.08±0.02 and 0.10±0.03 in the control group and was gradually enhanced with the increase of CoCl₂ concentration with the peak value at 200μmol/L CoCl₂,and the value for EPO and EPOR was 0.22±0.08,0.28±0.091 respectively.ConclusionHypoxia induced by CoCl₂ increase viability of RPE cells.Expression of EPO and EPOR in human fetal RPE cells has important effects on cells viability.更多还原