Vitreal macrophages express vascular endothelial growth factor in oxygen-induced retinopathy
Vitreal macrophages express vascular endothelial growth factor in oxygen-induced retinopathy
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DOI：
10.1046/j.1442-9071.2000.00226.x
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发表时间：
2000-02-01
影响因子：
4
通讯作者：
Gobé, G
中科院分区：
文献类型：
--
作者：
Naug, HL;Browning, J;Gobé, G
目的：探讨玻璃体巨噬细胞在氧源性视网膜病变(OIR)中发挥血管生成作用的可能性。为了建立人类早产儿视网膜病变(ROP)增殖期动物模型，在新生动物中建立了氧源性视网膜病变模型。材料和方法：新生小鼠出生后第7天，将其窝置于80~90%的氧气中5d，然后回到室内空气中，建立OIR模型。分别于出生后第7、12、15、17和20天处死幼鼠，然后用生理盐水冲洗、4%多聚甲醛和印度墨水进行灌流固定。眼球被摘除并整体安装，或被快速冷冻和冷冻切除。免疫组织化学染色显示巨噬细胞和血管内皮生长因子(VEGF)蛋白。所用一抗分别为抗F4/80和抗小鼠血管内皮生长因子。计数与玻璃体-视网膜界面密切相关的玻璃体巨噬细胞(距内界膜25um以内)。结果：与年龄匹配的对照组相比，氧处理组大鼠眼部巨噬细胞数量显著增加(P<；0.05)。巨噬细胞与玻璃体新生血管芽之间存在着密切的空间关系。结论：玻璃体巨噬细胞在OIR的发病机制中起重要作用，由此推测ROP的发生可能与氧有关。
Purpose: The possibility of vitreal macrophages playing an angiogenic role in oxygen-induced retinopathy (OIR) was investigated. Oxygen-induced retinopathy was produced in newborn animals with the purpose of modeling the proliferative phase of human retinopathy of prematurity (ROP).Materials and Methods: To produce OIR in neonatal mice, litters at postnatal day 7 were placed in 80-90% oxygen for a period of 5 days and then returned to room air. Pups were killed on days 7, 12, 15, 17 and 20 over the postnatal period and were perfusion-fixed using a saline wash-out, followed by 4% paraformaldehyde and then India Ink. Eyes were enucleated and either whole-mounted, or snap-frozen and cryosectioned. Immunostaining procedures were used to visualize macrophages and vascular endothelial growth factor (VEGF) protein. The primary antibodies used were anti-F4/80 and antimouse VEGF, respectively. Vitreal macrophages closely associated with the vitreo-retinal interface (within 25 mum of the inner limiting membrane) were counted. In situ hybridization procedures were used to analyse for the presence of VEGF mRNA transcript in vitreal macrophages.Results: Macrophage numbers were found to significantly increase (P < 0.05) in eyes from oxygen-treated animals compared with those from age-matched controls. A close spatial relationship was observed between macrophages and vitreal neovascular sprouts. In addition, vitreal macrophages were also found to transcribe and express VEGF in the oxygen-treated animals during the vasoproliferative phase.Conclusions: Our results raise the possibility that vitreal macrophages play a role in the pathogenesis of OIR and by inference, ROP.