高糖对体外培养兔视网膜Müller细胞AQP4表达的影响

Effect of high concentration glucose on AQP4 expression in rabbit retina Müller cell cultured in vitro

目的:观察体外培养兔视网膜M櫣ller细胞在高糖条件下水通道蛋白-4(AQP4)表达的变化。方法:采用体外培养的兔视网膜Müller细胞,分为正常对照组及高糖组(葡萄糖浓度:30、40和50mmol.L-1),分别培养1、3和5d;采用免疫细胞化学、流式细胞术及RT-PCR方法对Müller细胞AQP4的表达情况进行检测。结果:高糖组M櫣ller细胞AQP4表达的绿色荧光明显弱于正常对照组(P<0.01);流式细胞仪检测中波峰位置明显提前,AQP4表达强度较对照组明显减弱(P<0.01),且随着葡萄糖浓度的增高和作用时间的延长,AQP4表达强度逐渐减弱;血糖浓度50mmol.L-1培养3d时,RT-PCR半定量分析显示,AQP4 mRNA的表达比对照组明显降低(P<0.01)。结论:高糖能降低视网膜M櫣ller细胞AQP4的表达,且随着葡萄糖浓度的增高和作用时间的延长,AQP4的表达强度也逐渐减弱;高糖时AQP4的表达下降可能是机体对糖尿病视网膜病变的一种保护性反应。

Objective To observe the effects of high concentration glucose on aquaporin 4 （AQP4） expression in rabbit retina MOiler cell cultured in vitro. Methods The rabbit retinal Mailer cells were cultivated in vitro under the condition of 30, 40, 50 mmol · L^-1 high concentration glucose. The AQP4 expression was detected by immunocytochemical staining, FACS and RT-PCR after cultivated for 1, 3, and 5 d. Results Green fluorescence of AQP4 expression of Mailer cells in high glucose groups was weaker than that in control group （P〈0. 01）. FACS results indicated that the position of wave crest advanced obviously and AQP4 expression deseased （P〈0.01）. With the increasing of glucose concentration and prolongation of time, the extent of AQP4 expression was gradually weakened. RT-PCR results revealed that on the condition of 50 mmol · L^-1 glucose for 3 d, AQP4 mRNA expression was weaker than that in control group （P〈0. 01）. Conclusion High concentration glucose could desease the AQP4 expression of cultured retinal Maller cells. With the increasing of glucose concentration and prolongation of time, the AQP4 expression deseases gradually and it may be a protective response in diabetic retinopathy DR.