论文部分内容阅读
目的探讨低氧对大鼠视网膜Müller细胞低氧诱导因子(HIF)-1α和促红细胞生成素(EPO)蛋白表达的影响。方法体外传代培养大鼠视网膜Müller细胞,分为正常对照组(N组)、氯化钴浓度:50,100,150,200,300μmol/L(C50、C100、C150、C200、C300组),MTT检测不同浓度氯化钴对Müller细胞活力的影响。免疫细胞化学、细胞免疫荧光检测、Western印迹和酶联免疫吸附试验检测HIF-1α、EPO蛋白表达的情况。结果氯化钴浓度低于200μmol/L时,细胞活力不受氯化钴浓度的影响。从200μmol/L开始,细胞活力随氯化钴浓度增高而下降。氯化钴浓度高于50μmol/L时,HIF-1α、EPO蛋白免疫染色可见阳性表达。酶联免疫吸附试验检测,从50μmol/L开始HIF-1α蛋白的表达随氯化钴浓度升高而增加。Western印迹检测,EPO蛋白表达从50μmol/L开始增高,在150μmol/L时达到高峰,以后逐渐下降。结论低氧可引起细胞活性改变并诱导视网膜Müller细胞HIF-1α和EPO蛋白表达的变化。
Objective To investigate the effect of hypoxia on the protein expression of hypoxia inducible factor (HIF) -1α and erythropoietin (EPO) in retinal Müller cells in rats. METHODS: Rat retinal Müller cells were subcultured in vitro and divided into normal control group (N group), cobalt chloride concentration: 50,100,150,200,300μmol / L (C50, C100, C150, C200 and C300 group) Müller cell vitality. Immunocytochemistry, immunofluorescence assay, Western blot and enzyme-linked immunosorbent assay were used to detect the expression of HIF-1α and EPO protein. Results When the concentration of cobalt chloride was less than 200μmol / L, the cell viability was not affected by the concentration of cobalt chloride. From 200μmol / L, the cell viability decreased with the increase of cobalt chloride concentration. When cobalt chloride concentration was higher than 50μmol / L, positive expression of HIF-1α and EPO protein was observed by immunostaining. Enzyme linked immunosorbent assay showed that the expression of HIF-1α protein increased with the increase of cobalt chloride concentration from 50μmol / L. Western blot analysis showed that the expression of EPO protein increased from 50μmol / L to 150μmol / L, then decreased gradually. Conclusion Hypoxia can induce changes of cell activity and induce the expression of HIF-1α and EPO in retinal Müller cells.