改良墨汁明胶灌注法与胶原蛋白酶消化法在大鼠视网膜新生血管形态显示中的对比研究

目的　比较改良墨汁明胶灌注法与胶原蛋白酶消化法对大鼠视网膜新生血管形态显示的不同。方法　将鼠龄为７ｄ的ＳＤ大鼠１２只（２４眼）随机分成墨汁明胶灌注组（ｎ＝６）和胶原蛋白酶消化组（ｎ＝６），两组大鼠共同暴露于含体积分数８０％ ±２％氧浓度环境饲养５ｄ，然后回到正常氧环境饲养５ｄ。在第１８天时将一组幼鼠行心脏墨汁明胶灌注，另一组采用胶原酶消化法。取两侧眼球，一眼用于视网膜铺片，另一眼行组织切片观察突破内界膜的血管内皮细胞核数目，证实新生血管的增殖。部分切片做血管性血友病因子免疫组织化学染色。结果　改良墨汁明胶灌注法能很好显示大鼠视网膜新生血管。胶原蛋白酶消化法技术复杂，成片率低；联合共聚焦显微镜可以进行视网膜新生血管的三维结构观察。结论　改良墨汁明胶灌注显示大鼠视网膜新生血管优于胶原蛋白酶消化法，适宜广泛使用。

Comparative study between improved ink-gelatin perfusion and collagenase digestion in detection of rat retina neovessels

Objective To compare the difference of improved ink-gelatin perfusion and collagenase digestion in the detection of rat retinal neovessels. Methods Twelve 7-day-old SD rats were randomly divided into ink-gelatin perfused group ( 6 cases) and collagenase digestion group (6 cases). Two group of rats were exposed to 80% +2% oxygen for 5 days and then to the room air for another 5 days,respectively. At 18 days , one group pups the hearts were perfused with ink. the other adopt collagenase digestion and then their bilateral eye-balls were enucleated .one was used as retinal preparation to assess the change of retinal vessels.the other was used as histological section to observe and count the endothelial cell nucleus of new vessels extending from the retina into vitreous in ink-gelatin perfusion group. vWf immunohistochemistry were performed in the retinal section. Results Improved ink-gelatin perfusion could clearly display the rat retinal neovessels. The achievement ratio of collagenase digestion was lower with complexity of technology. Three-dimensional characteristics of retinal vessels could be observed with laser scanning confocal microscope. Conclusion Improved ink-gelatin perfusion is better than the collagenase digestion in the detection of rat retinal neovessels .which should be widely applied for the study of eye diseases.