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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 rh EPO+25 mmol/L葡萄糖),EG50组(4×104IU/L rh EPO+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。
Objective To investigate the effect of recombinant human erythropoietin on the function of retinal Müller cells cultured with high glucose in rats. Methods Rat retinal Müller cells were subcultured in vitro and divided into N groups (normal control), G25 group (25 mmol / L glucose), G50 group (50 mmol / L glucose), EG25 group (4 × 104IU / L rh EPO + 25 mmol / L glucose) and EG50 group (4 × 104 IU / L rh EPO + 50 mmol / L glucose). The cell viability was measured by MTT assay. Cell immunostaining and ELISA were used to detect the expression of GS protein in each group. Results The MTT assay showed that the cell viability in N group was higher than that in G25, G50, EG25 and EG50 groups, higher in EG25 group than in G25 group, and higher in EG50 group than in G50 group. Cell immunofluorescence staining showed that all the cells in each group had GS protein positive staining. Enzyme-linked immunosorbent assay showed that the expression of GS protein in N group was higher than those in high glucose group. The protein expression in EG25 group and EG50 group were higher than G25 group and G50 group respectively. Conclusion Recombinant human erythropoietin can increase retinal Müller cell activity and up-regulate the expression of GS protein in cultured rat with high concentration of glucose.