Upregulation of stromal cell-derived factor 1 (SDF-1) expression in microvasculature endothelial cells in retinal ischemia-reperfusion injury
Upregulation of stromal cell-derived factor 1 (SDF-1) expression in microvasculature endothelial cells in retinal ischemia-reperfusion injury
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DOI：
10.1007/s00417-008-0907-3
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发表时间：
2008-12-01
影响因子：
2.7
通讯作者：
Tang, Shibo
中科院分区：
文献类型：
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作者：
Lai, Pinghong;Li, Tao;Tang, Shibo
基质细胞衍生因子1 （SDF-1）是一种有效的趋化和血管生成因子，已被提出在新生血管的发展中发挥作用。在本研究中，我们探讨了SDF-1在视网膜缺血再灌注损伤大鼠模型中的表达，并探讨了视网膜微血管内皮细胞在该趋化因子产生中的可能作用。通过升高眼压至110 mm 60分钟，研究sd -1在缺血再灌注损伤大鼠视网膜中的表达规律。采用半定量RT-PCR检测视网膜再灌注6、12、24 h后SDF-1 mRNA的相对水平。免疫组织化学方法检测表达SDF-1的特异性病变。采用半定量RT-PCR方法检测缺氧条件下培养的人视网膜微血管内皮细胞（HRMEC）中SDF-1基因的表达。通过免疫细胞化学和荧光活化细胞分选分析SDF-1蛋白。SDF-1 mRNA在再灌注后6、12、24小时表达上调，12小时达到表达高峰。SDF-1阳性细胞最初出现在视网膜血管周围，并扩散到视网膜内层。缺氧可增强HRMEC中HIF-1和SDF-1 mRNA的表达。通过荧光激活的细胞分选证实，缺氧6小时后，HRMEC产生的SDF-1蛋白增加了320%。我们的研究结果表明，内源性SDF-1在遭受缺血损伤的视网膜微血管中上调，微血管内皮细胞是缺血视网膜中SDF-1生成的潜在贡献者。
Stromal cell-derived factor 1 (SDF-1) is a potent chemotactic and angiogenic factor that has been proposed to play a role in the development of neovascularization. In this study, we explored the expression of SDF-1 in a rat model of retinal ischemia-reperfusion injury and investigated the possible role of retinal microvasculature endothelium cells in generation of this chemokine.Expression patterns of SDF-1 were studied in retina suffering ischemia-reperfusion insult in Sprague-Dawley rats by elevating the intraocular pressure to 110 mm for 60 minutes. The relative level of SDF-1 mRNA in retinas following 6, 12 and 24 hours reperfusion was determined by semi-quantitative RT-PCR. Immunohistochemical methods were used to detect specific lesions expressing SDF-1. The gene expression of SDF-1 in cultured human retinal microvasculature endothelial cells (HRMEC) under hypoxia conditions was assessed by semi-quantitative RT-PCR. The SDF-1 protein was analyzed by immunocytochemistry and fluorescence-activated cell sorting.Upregulation of SDF-1 mRNA (at 6, 12, and 24 hours of reperfusion) was observed, with the expression peak occurring at 12 hours. SDF-1 positive cells appeared initially around the retinal vessels,which diffused into the inner retinal layers. Hypoxia enhanced the expression of HIF-1 and SDF-1 mRNA in HRMEC. The production of SDF-1 protein by HRMEC was increased up to 320% after 6 hours of hypoxia, as demonstrated by fluorescence-activated cell sorting.The results of our study indicate that endogenous SDF-1 is up-regulated in retinal microvasculature suffering ischemia insult, and that microvasculature endothelial cells are potential contributors for generation of SDF-1 in ischemic retina.