Wu Yu-Ching, Chang Chao-Yuan, Kao Alex, Hsi Brian, Lee Shwu-Huey, Chen Yau-Hung, Wang I-Jong
Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan.
Department of Chemistry, Tamkang University, Tamsui, New Taipei City, Taiwan.
PLoS One. 2015 May 15;10(5):e0126750. doi: 10.1371/journal.pone.0126750. eCollection 2015.
Retinopathy of prematurity, formerly known as a retrolental fibroplasia, is a leading cause of infantile blindness worldwide. Retinopathy of prematurity is caused by the failure of central retinal vessels to reach the retinal periphery, creating a nonperfused peripheral retina, resulting in retinal hypoxia, neovascularization, vitreous hemorrhage, vitreoretinal fibrosis, and loss of vision. We established a potential retinopathy of prematurity model by using a green fluorescent vascular endothelium zebrafish transgenic line treated with cobalt chloride (a hypoxia-inducing agent), followed by GS4012 (a vascular endothelial growth factor inducer) at 24 hours postfertilization, and observed that the number of vascular branches and sprouts significantly increased in the central retinal vascular trunks 2-4 days after treatment. We created an angiography method by using tetramethylrhodamine dextran, which exhibited severe vascular leakage through the vessel wall into the surrounding retinal tissues. The quantification of mRNA extracted from the heads of the larvae by using real-time quantitative polymerase chain reaction revealed a twofold increase in vegfaa and vegfr2 expression compared with the control group, indicating increased vascular endothelial growth factor signaling in the hypoxic condition. In addition, we demonstrated that the hypoxic insult could be effectively rescued by several antivascular endothelial growth factor agents such as SU5416, bevacizumab, and ranibizumab. In conclusion, we provide a simple, highly reproducible, and clinically relevant retinopathy of prematurity model based on zebrafish embryos; this model may serve as a useful platform for clarifying the mechanisms of human retinopathy of prematurity and its progression.
早产儿视网膜病变，以前称为晶状体后纤维增生症，是全球婴儿失明的主要原因。早产儿视网膜病变是由视网膜中央血管未能到达视网膜周边，导致周边视网膜无灌注，进而引起视网膜缺氧、新生血管形成、玻璃体积血、玻璃体视网膜纤维化和视力丧失。我们通过使用经氯化钴（一种缺氧诱导剂）处理的绿色荧光血管内皮斑马鱼转基因品系，在受精后24小时再给予GS4012（一种血管内皮生长因子诱导剂），建立了一种潜在的早产儿视网膜病变模型，并观察到治疗后2至4天，视网膜中央血管主干中的血管分支和新芽数量显著增加。我们使用四甲基罗丹明葡聚糖创建了一种血管造影方法，该方法显示有严重的血管渗漏，血管壁中的物质渗漏到周围的视网膜组织中。通过实时定量聚合酶链反应对幼虫头部提取的mRNA进行定量分析，结果显示与对照组相比，vegfaa和vegfr2的表达增加了两倍，这表明在缺氧条件下血管内皮生长因子信号增强。此外，我们证明了几种抗血管内皮生长因子药物，如SU5416、贝伐单抗和雷珠单抗，可以有效挽救缺氧损伤。总之，我们基于斑马鱼胚胎提供了一种简单、高度可重复且与临床相关的早产儿视网膜病变模型；该模型可能成为阐明人类早产儿视网膜病变机制及其进展的有用平台。
