氧诱导视网膜新生血管模型鼠视网膜水通道蛋白1的表达

Expression of aquaporin 1 in retinal neovascularization： experiment with mice

目的研究氧诱导视网膜新生血管模型鼠视网膜组织中水通道蛋白1（AQP1）表达的变化。方法高浓度氧诱导7d龄C57BL／6J幼鼠，建立视网膜缺血型病变动物模型。采用免疫组化法、逆转录聚合酶链反应（RT—PCR）和Western印迹法观察氧诱导模型组（12、15和17d）和正常对照组小鼠视网膜组织中AQP1的变化。结果部分12d小鼠和所有15d及17d的小鼠视网膜均有AQP1表达，且氧诱导模型组视网膜血管内皮细胞有AQP1表达；此外，17d氧诱导模型组小鼠视网膜的AQP1表达明显高于正常对照组（AQP1mRNA转录1．814±1．02V81．15±0．01；AQP1蛋白1．82±0．05V81．57±0．04）。结论AQP1可能参与视网膜新生血管的形成。

Objective To investigate the expression of aquaporin （AQP） 1 in retina during neovascularization. Methods Sixty C57BL/6J 7-day-old mice were randomly divided into 2 equal groups. The 30 mice of the model group were raised in 75%±2% oxygen for 5 days and then returned to condition with normal oxygen （room air） for 3 -5 days. 12, 15, and 17 days after birth （on the postnatal day 12, 15, and 17） 10 mice from each group were killed respectively. Immunohistochemistry was used on 6 eyes and RT-PCR and Western blotting were used to detect the mRNA and protein expression of AQP1 in 7 eyes respectively. Thirty mice were raised in normal oxygen as controls. Results The retinal neovascularization rate on P17 in the model group was 100%. AQP1 expression in retina was seen in 1 eye from the model and control groups each on P12, and in all mice of both groups on P15 and P17, especially in the lateral retina. AQP1 expression in retinal endothelial cells was seen in the model mice and not in the control mice. There were not significant differences in the AQP1 mRNA expression level on P12 and P15 between the 2 groups, however, the AQP1 mRNA expression level on P17 was significantly higher in the model group than in the control group （1.81±1.02 vs 1.15 ±0.01, P = 0.000）. AQP1 protein expression was seen on P12, P15, and P17 in both groups, and the AQP1 protein expression level at P17 of the model group was significantly higher than that of the control group （1.82±0.05 vs 1.57±0.04,P=0.000）. Conclusion AQP1 protein may play an important role in retinal neovascularization.