熊思齐 夏晓波 蒋 剑 孙 伟
Expression of netrin-1 m RNA in retina with experimental retinal neovascularization induced by oxygen
Xiong Siqi，Xia Xiaobo，Jiang Jian，Sun Wei．Department of Ophthalmology，Xiangya Hospital，Central South University，Changsha 410008，China
Abstract Objective It has been well—known that retinal neovascularization is associated to multiple factors．However，recent research showed that netrin一1 mRNA can promote the neovascularization．This study was to evaluate the expression of netrin-1 mRNA in retina in experimental retinal ne0vascularization induced by oxygen． M ethods Fifty one—week—old C57BL／6J mice were divided into control group and hyperoxic group．The mice of hyperoxic group were exposed to 75％ oxygen for 5 days and then returned to the room air to induce retinal neovascularization，and the mice in control group were raised in the environment of room air for the same time．The morphological observation of retinal neovascularization was performed using angiography by fluorescein dextran injection under the fluorescence microscope，and the new blood vessels were quantified after 5 days in room air(17一day-old)by counting the vascular epithelial cell nuclei protruding into viteous cavity using HE stain．RT—PCR
were used to examine retinal netrin一1 mRNA level in postnatal day 12，14 and 17 respectively． Results A lots of new blood vessels were seen surrounding the optic disk with irregular distribution and large area of non—perfused area and fluorescine leakage in the mouse retina in hyperoxic group．However，in the normal mouse retina，retinal blood vessels showed the network distribution under the fluorescence microscope．The neovascular nuclei protruding into viteous cavity was 1 7．2 ±2．6 in hyperoxic group．and no similar neovaseular nuclea was found in control group．No significant difference was found in retinal netrin一1 mRNA level in postnatal 12 days between hyperoxic group and control group but was significantly higher in postnatal 14·rday mice and 17--day mice compared with control mice(t=8．61，P=0．008；t=7．12，P=0．009)． Conclusion Netrin一1 mRNA 1evel increases in
hypoxia murine retina，which may play an important role in inducing retinal neovascularization．
Key words ne0vascularizati0n： retina： netrin-1
摘要 目的研究高氧诱导的视网膜新生血管模型鼠中轴突导向因子一i(Netrin一1)mRNA的表达差异。方法采
用高氧诱导的方法制作鼠视网膜新生血管模型；运用荧光造影视网膜铺片及视网膜切片苏木精一伊红染色观察视网膜新生血管的形态。于出生后第12、14、17 d取小鼠视网膜，采用RT-PCR测定Netrin一1 mRNA的表达水平。
结果 模型组视网膜铺片及组织切片可见大量视网膜新生血管形成。出生后12 d，模型组与正常组视网膜组织中Netrin一1 mRNA表达水平无明显差异；出生后14 d，模型组视网膜组织中Netrin一1 mRNA表达水平明显上调；出生后17 d模型组视网膜组织中Netrin一1 mRNA表达水平仍高于正常组。 结论模型鼠视网膜新生血管发生过程中，持续缺氧的视网膜组织可能从转录水平增加Netrin一1的表达，从而诱导视网膜新生血管的发生。
关键词新生血管；视网膜；轴突导向因子．1分类号 R 774 文献标识码A 文章编号1003-0808(2009)02—0105—04
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