重组人促红细胞生成素对高糖培养大鼠视网膜 Müller细胞的影响

摘要

Objective Present study was to investigate the effect of recombinant human erythropoietin on retinal Müller cells cultured in high glucose.Methods Serial subcultivated neonatal rats’ retinal Müller cells were divided into normal control group, G25(25mmol/Lglucose) group, G50(50mmol/Lglucose), EG25(4 ×104 IU/L recombinant human erythropoietin+25mmol/L glucose) group and EG50 (4 ×104 IU/L recombinant human erythropoietin +50mmol/Lglucose) group.MTT assay was applied to detect cell viability in five groups.The expression of glutamine synthetase was measured by immunofluorescence and enzyme linked immunosorbent assay.Results Cell viability of G25 group, G50 group, EG25 group and EG50 group was lower than that of normal group.Cell viability of EG25 group was higher than that of G25 group, and EG50 group was higher than that of G50 group.Cells expressed glutamine synthetase in five groups.The expression of glutamine synthetase in normal group was higher than that in other four groups where cells were cultured in high glucose. The expression of glutamine synthetase in EG25 group and EG50 group were higher than that in G25 group and G50 group re-spectively.Conclusion Recombinant human erythropoietin could increase the cell viability and the expression of gluta-mine synthetase in retinal Müller cells cultured in high glucose.%目的：探讨重组人促红细胞生成素对高浓度葡萄糖培养大鼠视网膜Müller细胞功能的影响。方法体外传代培养大鼠视网膜Müller细胞，分组为N组（正常对照），G25组（25 mmol／L葡萄糖），G50组（50 mmol／L葡萄糖），EG25组（4×104IU／L rhEPO＋25 mmol／L葡萄糖），EG50组（4×104IU／L rhEPO＋50 mmol／L葡萄糖），MTT检测各组细胞活性。细胞免疫染色、ELISA检测各组细胞GS蛋白表达的情况。结果 MTT检测示N组细胞活性高于G25组、G50组、EG25组和EG50组，EG25组高于G25组，EG50组高于G50组。细胞免疫荧光染色检测各组细胞均有GS蛋白阳性着染。酶联免疫吸附试验检测N组GS蛋白表达高于各高浓度葡萄糖培养组，EG25组、EG50组蛋白表达分别高于G25组和G50组。结论重组人促红细胞生成素可增加高浓度葡萄糖培养大鼠视网膜Müller细胞活性并上调GS蛋白的表达。