改良的可定量氧诱导法建立视网膜新生血管小鼠模型

A modified model of vascular proliferation in oxygen-induced retinopathy in mouse

目的建立简单的、成模效率高的可定量氧诱导视网膜新生血管C57BL/6小鼠模型。方法89只C57BL/6乳鼠随机分为3组,分别用传统法和两种改良法建立模型,12只作为正常对照组。比较各组母鼠死亡率、乳鼠存活率和成模率。随机取1只眼行视网膜石蜡切片苏木精-伊红染色,另1只眼行ADP酶法视网膜铺片,进行染色观察和定量分析。结果与传统法相比,两种改良法母鼠死亡率低,乳鼠存活率和成模率较高。改良Ⅰ组视网膜平均每张切片突破内界膜增生细胞数和无灌注区相对面积比其他两组明显增多,差异有统计学意义(P<0.05)。改良Ⅱ组与改良Ⅰ组比较,乳鼠存活率高,细胞增生和无灌注区明显且程度适中。结论两种改良法简单、稳定且高效,均能构建典型的可定量氧诱导视网膜新生血管模型,改良法Ⅱ是适合视网膜新生血管发生机制及药物干预研究的一种方法。

Objective Oxygen-induced retinopathy（OIR） mouse model was most frequently used for retinal neovascular research in the world. This study was to establish a simple, effective and quantitative model of vascular proliferation in OIR mouse. Methods Eighty-nine C57BL/6 mice were divided into conventional feeding group（mice was rose for 5 consecutive days in oxygen cabin） ,modified Ⅰ methods（mice was rose at a 6-hour interval per day for 5 consecutive days in oxygen cabin） and modified Ⅱ method （mice was rose at a 12-hour interval per day for 5 consecutive days in oxygen cabin） to establish OIR model,and other 12 normal mice were as controls. In postnatal 17 days （ P17 ） , the mortality rate of breast-feeding mice and survival rate of neonatal mice were compared among different groups. The proliferative angiogenesis response was evaluated quantitatively by counting the cells extending from retina inner limiting membrane （ILM） into vitreous. The retinal flat mounts were prepared and ADPase histochemical technique was used to assess the retinal non-perfusion area. Results Compared to conventional group,modified Ⅰ group and modified Ⅱ group showed the characteristics of lower breast-feeding mice mortality rate and higher P17 neonatal mice survival rate （47.06% , 65.52% , 88.46% respectively） （X^2 = 2.16, P ≈ 0.14;X^2 = 11.1, P 〈 0.01 ） , and that of modified Ⅱ group was higher than that modified Ⅰ group （X^2 = 3.99, P 〈 0.05 ） . The mean proliferative cells and non-perfusion area of retina in modified Ⅰ group were improved markedly,showing a significant difference in comparison with conventional and modified Ⅱ model groups （P 〈 0. 05 ）. The P17 mice survival rate in modified Ⅱ group was higher than that of modified Ⅰ group. Compared with conventional groups,the extending proliferative cells from ILM into vitreous was obvious and midrange in modified Ⅱ group,showing the higher modeling successful rate in modified Ⅰ and Ⅱ groups. The retinal non-perfusion area in modified Ⅰ group was significant increased in comparison with conventional group and modified Ⅱ group（P 〈 0.05）. Conclusion The two modified models can be created with simple,reproducible,stable features. Modified Ⅱ modeling approach is proved to be suitable for the study of retinal angiogenesis and drug intervention.