氧诱导视网膜病变小鼠模型的建立与评价

Establishment and evaluation of the mice model of oxygen induced retinopathy

目的:建立氧诱导视网膜病变(OIR)小鼠模型,并对模型进行整体评价。方法:7日龄清洁级C57BL/6J小鼠随机分为实验组(高氧组)与对照组(空气组),实验组置于体积分数为75%高氧环境中饲养5d后回到空气中继续饲养,对照组则始终在空气环境中饲养。两组小鼠分别于生后第7(P7)、12(P12)、14(P14)、17(P17)、22(P22)、25(P25)及30(P30)天处死取材,观察视网膜病变。视网膜铺片ADP酶染色了解视网膜血管形态学改变;HE染色计数突破视网膜内界膜的内皮细胞核数目,定量反映视网膜血管的增生情况;VG(Van Gieson)染色及I/III型胶原纤维免疫组织化学染色判断视网膜后期纤维化情况。结果:①成功建立了OIR小鼠模型;对视网膜血管增生定量分析显示实验组小鼠P17平均每只眼球每张切片中新生血管内皮细胞核数目达(32.88±5.843)个,而对照组不足1个,差异具有统计学意义(P<0.000 1)。②探索了OIR小鼠视网膜晚期纤维化情况:视网膜新生血管P17后逐渐消退;视网膜组织VG染色及I/III型胶原纤维免疫组织化学染色结果皆为阴性。结论:OIR小鼠模型较好地复制了临床早产儿视网膜病(ROP)中血管阻塞、新生血管形成等急性病变,但对严重ROP中的视网膜纤维化、牵引性视网膜脱离未能呈现。

Aim：To establish the mice model of oxygen induced retinopathy（OIR）,and evaluate the model overall.Methods： Seven-day-old C57BL/6J mice of clean grade were randomly random divided ized into oxygen-induced retinopathy group and control group.In oxygen-induced retinopathy group,mice were exposed to 75% oxygen for 5 days and then to room air;in control group,mice were raised in room air.Eye samples of mice from each group were excised at postnatal 7 days（Postnatal days 7,to express P3,the same below）,P12,P14,P17,P22,P25,and P30 respectively.The morphologic changes of retinal vessels were estimated by observing the vascular pattern with ADPase staining retina flatmounts,and quantitated by counting the number of new vascular cell nuclei extending into the internal limiting membrane by HE stain,using VG stain and immunohistochemical stain to assess the interstitial fibrosis of the retina.Results： ①An oxygen induced retinopathy（OIR） in mice was successfully induced following 5-day exposure to 75% oxygen,and the quantitative results of the retinal neovascularization showed： there was a mean of 33 neovascular nuclei per cross-section extending into the vitreous in hyperoxia compared to less than 1 nuclei in the normoxia control（P〈0.000 1）.②Exploring the situation of the retina fibrosis： The retinal neovascularization began to fade after P17;the results of VG stain and immunohistochemistry stain were all negative.Conclusion：The mice model of OIR duplicated preferably the acute pathological changes of retinopathy of prematurity（ROP） such as angiemphraxis and retinal neovasculatization,but it not present the serious pathological changes such as retinal fibrosis and retinal detachment of ROP.