色素上皮衍生因子对鼠视网膜新生血管形成的保护作用

目的本课题通过小鼠视网膜新生血管动物模型观察玻璃体内注入色素上皮衍生因子(PEDF)对视网膜新生血管形成的影响,并探讨其治疗作用。方法选取鼠龄为7天的健康昆明小鼠28只(56只眼),随机分为4组:正常组、高氧组、PEDF干预组和对照生理盐水组,每组7只。正常对照组鼠在正常空气环境中饲养,其余3组鼠置于氧箱中,建立高氧诱导的视网膜新生血管动物模型。正常组鼠不做任何处理,PEDF干预组幼鼠在出生后第12天向玻璃体腔内注射PEDF 1μl,而对照生理盐水组注射相同体积的生理盐水。各组小鼠均在出生后第17天处死,摘除眼球制作标本,进行组织病理学观察。在光学显微镜下观察并计数突破视网膜内界膜的血管内皮细胞细胞核数目。采用CD31分子免疫组化方法鉴别视网膜内界膜与血管内皮细胞。结果正常对照组标本HE染色未见突破内界膜的内皮细胞核,高氧组和对照生理盐水组标本可见较多的突破内界膜的内皮细胞核,与正常组比较差异具有显著性(P<0.05),PEDF干预组的数目明显少于高氧组和对照生理盐水组,差异均具有显著性(P<0.05),且未见明显药物毒性反应。CD31分子免疫组化染色证实突破内界膜的细胞为血管内皮细胞。结论玻璃体内注入一定剂量的PEDF,能够有效抑制高氧诱导下的小鼠视网膜新生血管形成,PEDF有望成为防治血管增生性视网膜病变的一种有效的方法。

Study on effect of pigment epithelial-derived factor in the protection for retinal neovascularization in mice

Objective To study the effect and the therapeutically role of pigment epithelial-derived factor(PEDF) in the protection for retinal neovascularization in mice.Methods ①Twenty-eight one-week-old Kunming mice were randomly divided into four groups: normal group,hyperxia group,PEDF group and 0.9%NaCl injected control group.The normal group was fed in the normal environment.The other groups were exposed to 75% oxygen to establish a model of retinal neovascularization.Mice in normal group did not receive any treatment.Mice in PEDF group were given an intravitreal injection of 1μl of PEDF and the same volume of 0.9%NaCl was injected into 0.9%NaCl group mice as a control.②All mice were killed in 17-day old.The eyes were enucleated for histological examination.③the number of nuclei of endothelial cell extending from retinal internal limiting membrane was counted by optics microscope.the endothelial cell and retinal internal limiting membrane were identified by CD31 Immunocytochemical method.Results no nuclei of endothelial cell extending from retinal internal limiting membrane was found in normal group,while some were found in hyperxia group and 0.9%NaCl injected control group.there was significant difference between them(P<0.05).But less nuclei were found in the PEDF group.there was significant difference between them(P<0.05).and no side effect was found.the cells extending from retinal internal limiting membrane were confirm to be endothelial cell by CD_(31) Immunocytochemical method.Conclusion Intravitreal injection of PEDF may suppress the retinal neovascularization in the mice oxygen model which suggests that intravitreal injection of PEDF may have potential therapeutic benefits in retinal vascular disease.