兔眼IPE与RPE细胞膜表面K-NPPase酶组织化学的电镜图像分析  

Microscopic image analysis of the K-NPPase enzyme histochemistry on cellular membrane of rabbit's IPE and RPE

作　　者：王海瑛[1] 刘哲丽[2,3] 

机构地区：[1]中国辽宁省沈阳市第四人民医院眼科沈阳市眼病研究所,110031 [2]中国辽宁省沈阳市,110001 [3]中国医科大学第一临床学院眼底病中心

出　　处：《国际眼科杂志》2006年第2期365-368,共4页International Eye Science

摘　　要：目的:为了探讨自体虹膜色素上皮细胞(IPE)能否代替视网膜色素上皮细胞(RPE)移植到视网膜下腔执行物质转运功能,对兔眼IPE与RPE细胞膜表面的Na+,K+-ATPase采用K-NPPase酶组织化学技术进行定位和分析比较。方法:家兔12只,取IPE和RPE组织固定,配制孵育液后进行K-NPPase酶组化孵育,制成电镜标本,采集图像,利用Metamorph/DP10/BX51系统进行分析。随机选取IPE细胞膜以及RPE细胞顶端相邻K-NPPase颗粒两点间的距离进行测量。计算每组样本的均值±标准差(x±s),进行成组设计材料的t检验。结果:有色素组与无色素组汇总后的IPE细胞膜上K-NPPase颗粒的距离平均值40.16±1.19nm,RPE顶端微绒毛细胞膜上K-NPPase颗粒的距离平均值为41.07±1.78nm,两者之间的差异无显著性意义(0.5>P>0.2)。结论:兔眼IPE细胞膜表面与RPE细胞顶端微绒毛膜表面均有Na+,K+-ATPase分布,采用K-NPPas酶组化电镜观察到的兔眼IPE细胞与RPE细胞可能具有相似的物质转运功能。AIM： To investigate whether IPE cells could replace RPE cells to transplant into subretinal space and exert material transportation function, the distribution of K-NPPase stained grains on the cellular membrane of rabbit＇s IPE and RPE by electronic image analysis was observed. METHODS： 12 rabbits were choosed as experimental animals. The eyes were enucleated immediately after death, and fixed by injecting 40g/L paraformaldyhyde-Sg/L glutaraldehyde containing 60g/L sucrose into the eyeballs and immersed in room temperature for 45min. The eyeballs were opened and IPE and RPE tissues were cut into 2mm × 4mm sections for incubation. Ultrathin sections were observed under transmission electron microscope. Adopted Metamorph/ DPIO/BXS1 system was used to analyse each transmission electron microscope photographs. The distance between two adjacent K-NPPase on the IPE cellular membrane or RPE cells＇ summit was detected. Mean and standard deviation （ ^-x±s） were calculated. Comparison between the IPE group and the RPE group was performed using t test. RESULTS： The mean distance between two adjacent K-NPPase on the IPE cellular membrane or RPE cells＇ summit was similar （0.5〉P〉0.2）. CONCLUSION： K-NPPase enzyme grains are distributed on both IPE and RPE cellular membrane, which indicates there are enough distribution of Na^＋,K^＋-ATPase on both IPE and RPE cellular membrane.

关 键 词：虹膜色素上皮细胞 视网膜色素上皮细胞 钾依赖性对硝基苯磷酸酶 钠钾ATP酶 物质转运功能 

分 类 号：R77[医药卫生—眼科]