摘要
目的 观察可溶性fms-样酪氨酸激酶受体-1（sFlt-1）不同基因片段对小鼠视网膜新生血管（RNV）的抑制作用.方法 构建携带sFlt-1（2～3）、（2～4）免疫球蛋白样区域编码基因的重组慢病毒sFlt-1（2～3）和sFlt-1（2～4）.鼠龄7 d的C57/6J小鼠96只,数字表法随机分为4组,每组24只.1组为正常对照组;2组为模型对照组;3组为sFlt-1（2～3）组;4组为sFlt-1（2～4）组.2、3、4组置于（75±2）%氧浓度环境,出生后12 d再置于正常空气下饲养,并分别玻璃体腔注射空病毒、慢病毒sFlt-1（2～3）和sFlt-1（2～4）各1μl.出生后17 d,荧光素灌注造影行视网膜铺片,观察视网膜血管改变;视网膜切片行苏木精-伊红染色,计数小鼠RNV细胞核数;蛋白质免疫印迹法（Western blot）检测视网膜血管内皮生长因子（VEGF）和VEGF受体-2含激酶插入区受体/胎肝激酶-1（KDR/Flk-1）的蛋白表达.结果 出生后17 d,与2组比较,3、4组视网膜荧光渗漏面积、RNV、突破内界膜（ILM）的血管内皮细胞核数均明显减少（P〈0.01）;VEGF蛋白表达无明显变化（P〉0.05）,KDR/F1k-1蛋白表达明显下降（P〈0.01）.结论 sFlt-1（2～3）和sFlt-1（2～4）基因片段能明显抑制氧诱导小鼠RNV的形成,sFlt-1（2～3）效果更为显著.
Objective To investigate the inhibitory effects of fms-like typrosine kinase receptor sFh-1 on retinal neovascularization （RNV）. Methods Recombinant lentivirus sFh-1 （ 2-3 ） and sFh-1 （ 2-4 ）expressing the sFh-1 （2-3） and （2-4） immunoglobulin-like regions of sFh-1 were constructed. 96 seven-dayold C57/6J mice were randomly divided into 4 groups with 24 mice in each group. Group 1 ： normal control;group 2： experimental control; group 3： sFlt-1（2-3）; group 4： sFlt-1（2-4）. The mice in group 2-4 were exposed to hyperoxia with （75±2）% O2 for 5 days and then returned to normoxia with 21% O2 ; the mice received an intravitreal injection with 1 μl virus of empty vector, sFlt-1（2 3）, or sFlt-1（2-4）, respectively. Five days later, all mice underwent perfusion fluorecein angiography and retinal wholemont was made to observe the changes of retinal vessels; retinal sections were stained by hematoxylin and eosin and RNV endothelium cell nucleus were counted; vascular endothelial growth factor （VEGF） and VEGF receptor-2 （KDR/Flk-1） protein were measured by Western blot. Results Seventeen days after birth, the retinal area of fluorescein leakage and RNV, RNV nucleus which breaking through inner limiting membrane in group 3 and 4 were smaller or less than that in group 2 （P〈0.01） ; while VEGF protein didn＇t changed much （P〈 0.05）the expression of KDR/Flk-1 decreased significantly （P〈0.01）. Conclusion sFlt-1（2 3）and sFlt-1 （2-4）can inhibit the formation of oxygen-induced RNV, the former virus has a better effect.
出处
《中华眼底病杂志》
CAS
CSCD
北大核心
2010年第3期219-222,共4页
Chinese Journal of Ocular Fundus Diseases
基金
上海市登山计划（064119543）
关键词
视网膜新生血管化/预防和控制
血管内皮生长因子受体1/治疗应用
基因转移技术
动物实验
Retinal neovascularization/prevention ＆ control
Vascular endothelial growth factor receptor-1/therapeutic use
Gene transfer techniques
Animal experimentation
