摘要
目的:通过高氧诱导的小鼠视网膜新生血管动物模型玻璃体内注入GM6001观察对视网膜PEDF表达的影响。方法:选取鼠龄为7d的SPF级C57BL/6J小鼠48只(96眼),随机分为4组:正常组、高氧组、GM6001干预组和磷酸盐缓冲液对照组,每组12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。在出生后第12dGM6001干预组幼鼠向玻璃体腔内注射GM60011μL(100μmol/L),而磷酸盐缓冲液对照组注射相同体积的磷酸盐缓冲液,正常组和高氧组不做处理。各组小鼠均在出生后第17d处死。摘除左眼球制作石蜡标本,分别用抗色素上皮衍生因子(PEDF)和抗CD34抗体作免疫组织化学标记视网膜切片进行组织病理学观察;右眼剥离视网膜提取蛋白,采用ELISA法分析PEDF以及基质金属蛋白酶(MMPS)在视网膜中的含量。结果:17d龄时,正常组和GM6001组小鼠在视网膜各层可见PEDF阳性表达产物,其中以神经节细胞层及神经纤维层表达为著,为高表达,对照组与高氧组小鼠,17d龄时表达显著减少,只在神经节细胞层有较高表达,其余各层表达极少见。MMP-2,MMP-9,PEDF的含量GM6001组和PBS对照组及高氧组之间在统计学上均有差异P<0.01GM6001组和正常组之间没有统计学差异P>0.05,高氧组和PBS对照组没有统计学差异P>0.05。结论:玻璃体内注入一定剂量的GM6001通过抑制基质金属蛋白酶2和基质金属蛋白酶9能够有效增强高氧诱导下的小鼠视网膜PEDF的表达并使新生血管的增长受到抑制。
AIM： To observe the expression of pigment epitheliumderived factor （PEDF） in the retina of mice by high oxygen-induced retinal neovascularization with injecting GM6001 in vitreous cavity.
METHODS： Forty-eight C57BL/6J mice （96 eyes） aged 7 days in SPF were randomly divided into four groups： normal group, high-oxygen group, GM6001 intervention group and phosphate buffer control group, with 12 mice in each. The normal group was fed in normal air, and the three groups were placed in oxygen boxes to establish high oxygen-induced retinal neovascularization animal models, at 12 days after birth, GM6001 intervention group was injected GM6001 1μL into the vitreous cavity, and phosphate buffer control group was injected with the same volume of phosphate buffer. No management was done on Normal or high-oxygen groups. The mice were executed at 17 days after birth. Their left eyes were removed for specimens, and anti-PEDF and anti-CD34 antibodies were used respectively for immunohistochemical biopsy pathology observation. Retinal tissue protein was extracted from their right eyes to analyzing the content of MMP-2, MMP-9 and PEDF in the retina with ELISA.
RESULTS.. In normal and GM6001 group, at 17-day-old, PEDF positive expression was appeared in every retinal layer, especially in ganglion cells and nerve fiber layer. In high-oxygen and control group, at 17-day-old, PEDF expressed a significant reduction, only a higher expression in the ganglion cells and nerve fiber layer. At the remaining floors, the expression could rarely be seen. The expression of MMP-2, MMP-9 and PEDF was statistically significant in GM6001, PBS control and high-oxygen group （P 〈 0.01）. That in GM6001 and normal group had no statistical signify （ P〉 0.05）. And in high - oxygen and PBS control group, the expression of MMP-2, MMP-9 and PEDF was not statistical significant （P〉 0.05）
CONCLUSION： In the oxygen-induced retinopathy in mice,injection of GM6001 1μ LI into the vitreous cavity can inhibit MMP-2 and MMP-9, effectively strengthen the expression of PEDF, and inhibit the growth of neovascularization.
出处
《国际眼科杂志》
CAS
2008年第7期1337-1340,共4页
International Eye Science
