摘要
目的：观察高浓度葡萄糖对体外培养的大鼠视网膜Müler细胞P2Y2受体表达的影响。方法：用改进的酶消化法纯化培养并用胶质原纤维酸性蛋白（GFAP）和谷氨酰胺合成酶（GS）鉴定大鼠Müller细胞。分别在对照组（5.5mmol/L）与高糖组（10，25，50mmol/L）中孵育24h和72h，通过免疫荧光及荧光强度半定量分析检测Müller细胞中P2Y2受体表达的变化。结果：纯化培养到第3-4代的Müller细胞，经鉴定纯度大于90%，表达GFAP和GS。在24h，10，25，50mmol/L组的荧光强度分别为31.05±7.06，59.17±11.42和84.11±12.72，与对照组（26.60±5.15）相比较均明显增高（P〈0.01）。而在72h，仅25和50mmol/L组的荧光强度明显强于对照组（P〈0.01）。结论：高浓度葡萄糖上调大鼠视网膜Müller细胞P2Y2受体的表达，提示这一受体可能参与糖尿病视网膜病变过程。
AIM：To observe effects of high glucose on P2Y2 receptor of rat Müller cells in vitro.METHODS：The rat Müller cells were cultured,purified by a modified enzyme-digesting method and identified by glial fibrillary acidic protein（GFAP）and glutamine synthetase（GS）.The cells were exposed to various concentrations of 5.5mmol/L（normal control）,10,25 and 50mmol/L glucose（high glucose）for 24 hours and 72 hours.The immunofluorescence staining and its intensity were then applied to measure the changes of P2Y2 receptor of rat Müller cells.RESULLS：The Müller cells were purified from mixed cells and cultured 3-4 passages with a purity of more than 90% and expression of GFAP and GS.The fluorescence intensity was increased significantly in the cells exposed to high concentrations of glucose（10,25 and 50mmol/L）for 24 hours with the values of 31.05±7.06,59.17±11.42 and 84.11±12.72 respectively,compared with that of controls（26.60±5.15,P〈0.01）.However,at 72 hours,the cells that only exposed to higher glucose of 25 and 50mmol/L showed more intensive fluorescence than normal controls（P〈0.01）.CONCLUSION：The results showed that high concentra-tions of high glucose could induce up-regulation of P2Y2 receptor of rat Müller cells,indicating that this receptor may be involved in the process of diabetic retinopathy.
出处
《国际眼科杂志》
CAS
2008年第4期717-720,共4页
International Eye Science
