激光损伤后不同时间点鼠视网膜中水通道蛋白-1的表达

目的：采用免疫组化、荧光定量PCR法观察激光损伤后不同时间鼠视网膜中水通道蛋白?1（ aquaporin?1，AQP?1）的分布及其mRNA表达的变化。方法 Brown Norway（ BN）大鼠激光损伤视网膜后，脊椎脱臼法处死，摘除眼球，4％福尔马林固定，石蜡包埋。平行视神经矢状连续切片，HE染色、免疫组化染色和阴性对照。用已知有AQP?1表达的正常大鼠眼球眶周组织中的血管切片为阳性对照。另取出大鼠眼视网膜组织，行实时定量PCR检测。结果免疫组化切片显示，褐色粗颗粒出现在大鼠眼组织细胞中为阳性，无则为阴性。实时荧光定量PCR结果显示，正常组AQP?1 mRNA表达量为：（1．15±0．01），激光损伤即刻：（1．10±0．01）；12 h：（1．10±0．03）；24 h：（1．16±0．01）；72 h：（1．13±0．01）。结论 AQP?1在眼内阳性表达于多个与水代谢有关的部位，视网膜上AQP?1 mRNA的表达在激光损伤后呈现动态变化，激光损伤初期呈上调，损伤后期呈下调表达。

Expression of aquaporin-1 in rat retina at different time points after laser injury

Objective Using immunohistochemistry method, fluorescence quantitative polymerase chain reaction (PCR) to observe the AQP?1 distribution at rat retina and mRNA expression changes in different time after laser injury. Methods The Brown Norway ( BN) rats were killed by spinal dislocation method after laser damaged, removed the eyeball, fixed in 4% formalin and embed?ded in paraffin. Parallel optic sagittal serial sections, HE staining, immunohistochemical staining and negative control were performed. Use the vascular slice in normal rat eyes?periorbital tissue, in which AQP?1 expression was known as a positive control. Remove the eye retina tissue of rats and check it with line real?time quantitative PCR. Results AQP?1 immunohistochemical examination showed brown coarse particles in rat ocular tissues and cells as positive, no as negative. Real?time PCR results showed normal AQP?1 mRNA expression level:1.15±0.01;laser damage instantly:1.10±0.01;12 hours:1.10±0.03;24 hours:1.16±0.01;72 hours:1.13±0.01. Conclusion AQP?1 is expressed in many parts of the water metabolism, and the expression of AQP?1 mRNA in the retina after laser injury shows dynamic changes, which of the initial laser damage was up, and the latter was down.