早产儿视网膜病变小鼠模型视网膜PEDF的表达和血清PEDF水平的变化及其意义  

Expression of PEDF in Retina and Changes of Serum PEDF in Mouse Model of Retinopathy of Prematurity

作　　者：张梅虹[1] 章学英[1] 杜鹃[1] 

机构地区：[1]复旦大学附属金山医院儿科,上海201508

出　　处：《中国临床医学》2013年第4期484-487,共4页Chinese Journal of Clinical Medicine

摘　　要：目的:研究高氧诱导早产儿视网膜病变(retinopathy of prematurity,ROP)小鼠模型视网膜中色素上皮衍生因子(pigment epithelium derived factor,PEDF)的表达和血清PEDF水平变化及其意义。方法:选择7日龄健康C57BL/6J新生小鼠112只,按照随机数字表分为2组,每组56只。模型组在75%高氧密闭容器中饲养5 d,然后返回到空气环境中饲养;对照组始终在空气环境中饲养。采用ADP酶组织化学法观察小鼠视网膜铺片中血管形态的改变;采用HE染色法计数视网膜切片中突破内界膜的新生血管内皮细胞核数目,定量观察视网膜血管增生情况;采用免疫组织化学法检测小鼠视网膜PEDF蛋白的表达;采用酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)法检测小鼠血清PEDF水平。结果:模型组在生后第12天的视网膜铺片显示中央大片无灌注区,在生后第17天的视网膜铺片显示大量新生血管形成。模型组小鼠在生后第17天和第22天时可见较多突破内界膜长入玻璃体的血管内皮细胞核,与对照组比较差异有统计学意义。第17天时:(69.4±9.3)比(1.5±1.2);第22天时:(76.0±8.4)比(1.3±1.1),P均<0.01)。模型组第12天时视网膜PEDF阳性细胞率显著高于对照组[(11.5±4.5)%比(4.4±2.4)%,P<0.01],第17天时则显著低于对照组[(6.1±4.0)%比(14.1±5.8)%,P<0.01]。模型组小鼠血清PEDF水平较低,第17天和第22天时显著低于对照组[第17天时:(148.9±71.9)ng/L比(232.2±84.2)ng/L,P<0.05;第22天时:(66.1±9.3)ng/L比(163.8±41.5)ng/L,P<0.01]。结论:ROP模型小鼠视网膜PEDF在高氧时过度表达,低氧时表达减少。ROP模型小鼠血清PEDF水平的变化滞后于视网膜PEDF的改变,不能更早地预测ROP的发生。Objective：To study the expression of pigment epithelium derived factor（PEDF）in retina and serum PEDF level in the hyperoxia-induced mouse model of retinopathy of prematurity（ROP）.Methods：A total of 112 seven-day-old C57BL/6J mice were randomly divided into two groups.The model group（n=56）was exposed to 75% hyperoxia for 5 days and then to room air to produce ROP model.The control group（n=56）was exposed to room air.ADP enzymohistochemistry staining method of retinal slices was used to observe the change of retinal neovascularization.The retinal neovascularization was quantified by counting the number of the endothelial nuclei extending into the internal limiting membrane.The immunohistochemistry method was used to study the expression of PEDF in retina.The level of serum PEDF was measured by enzyme-linked immunosorbent assay（ELISA）.Results：In the model group,after 5 day exposure to hyperoxia,central perfusion decreased obviously,and a large number of neovessels had formed.At the 17th day and 22th day,compared with the normal group,there were more neovascular endothelial nuclei breaking the retinal tissue membrane in the model group（at the 17th day,69.4±9.3 vs.1.5±1.2;at the 22th day,76.0±8.4 vs.1.3±1.1,P〈0.01）.Immunohistochemistry results showed that in retinal tissues of the model group,PEDF was strongly expressed at the 12th day[（11.5±4.5）% vs.（4.4± 2.4）%,P〈0.01],but it was weakened at the 17th day[（6.1±4.0）%vs.（14.1±5.8）%,P〈0.01].ELISA showed that the level of serum PEDF in the model group was lower,especially at the 17th day and at the 22th day[at the 17th day,（148.9±71.9）ng/L vs.（232.2±84.2）ng/L,P〈0.05;at the 22th day：（66.1±9.3）ng/L vs.（163.8±41.5）ng/L,P〈0.01].Conclusions：In the hyperoxia-induced mouse model of ROP,PEDF is overexpressed in hyperoxia,and weakened in the room air.In the hyperoxia-induced mouse model of ROP,the change of serum PEDF level lagged behind the change of retinal PEDF.

关 键 词：早产儿视网膜病变 色素上皮衍生因子 新生血管化 

分 类 号：R722.6[医药卫生—儿科]