Soluble EphB4 regulates choroidal endothelial cell function and inhibits laser-induced choroidal Neovascularization
Soluble EphB4 regulates choroidal endothelial cell function and inhibits laser-induced choroidal Neovascularization
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DOI：
10.1167/iovs.05-0502
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发表时间：
2005-12-01
影响因子：
4.4
通讯作者：
Hinton, DR
中科院分区：
文献类型：
--
作者：
He, SK;Ding, Y;Hinton, DR
目的。本研究旨在探讨可溶性单体EphB4胞外区(SEphB4)对脉络膜内皮细胞(CEC)迁移和新生血管形成的影响以及对激光诱导的脉络膜新生血管(CNV)的影响。用改良的Boyden小室实验评价sEphB4(0.5~3 mU g/m L)对CEC迁移的影响。在胶原胶中检测CEC培养中的管状形成。用激光光凝法诱导大鼠新生血管形成。第14天，通过荧光素血管造影(FA)、异凝素-B4标记的平板共聚焦体积分析和CNV膜的组织学检查，评估sEphB4对CNV发育的影响。结果：Western印迹分析显示CEC细胞表达EphB4和EPhinB2。大鼠眼部免疫组织化学切片显示脉络膜内皮细胞EphB4和EPhinB2均呈免疫反应阳性。SEphB4减少CEC对血管内皮生长因子的迁移(P<；0.01)。同样，sEphB4以剂量依赖的方式抑制CEC管的形成。在激光诱导的CNV膜内的血管通道上检测到EphB4和较小程度的EPhinB2。玻璃体内注射sEphB4可抑制激光诱导的新生血管形成。CNV膜渗漏分数降低(P<；0.05)，膜体积缩小(P<；0.05)。组织学分析显示经sEphB4处理的膜血管减少。结论重组可溶性单体EphB4在体外和体内均对脉络膜血管生成有抑制作用。它作为一种新的治疗CNV的方法的潜力值得进一步评估。
PURPOSE. The purpose of this study was to evaluate the effect of a soluble monomeric form of the EphB4 extracellular domain (sEphB4) on choroidal endothelial cell (CEC) migration and tube formation and on experimental laser-induced choroidal neovascularization (CNV).METHODS. EphrinB2 and EphB4 expression in CECs was investigated by Western blot analysis and immunohistochemistry. Effects of sEphB4 ( 0.5 - 3 mu g/mL) on CEC migration were evaluated with a modified Boyden chamber assay. Tube formation was assayed in CEC cultures in collagen gel. CNV was induced in rats by laser photocoagulation. The effects of intravitreal injection of sEphB4 on CNV development were evaluated at day 14 by fluorescein angiography ( FA), confocal volumetric analysis of isolectin-B4 labeled flatmounts, and histologic examination of CNV membranes.RESULTS. CEC cells express both EphB4 and EphrinB2, according to Western blot analysis. Immunohistochemical sections of rat eye showed immunoreactivity for both EphB4 and EphrinB2 in the choroidal endothelium. sEphB4 reduced CEC migration in response to vascular endothelial growth factor ( P < 0.01). Similarly, sEphB4 inhibited CEC tube formation in a dose-dependent manner. EphB4, and to a lesser extent EphrinB2, were detected on vascular channels within laser-induced CNV membranes. Intravitreal injection of sEphB4 inhibited laser-induced CNV formation. CNV membranes showed a reduction in leakage score ( P < 0.05), and membrane volumes were reduced in size ( P < 0.05). Histologic analysis revealed that vascularity was reduced in sEphB4-treated membranes.CONCLUSIONS. Recombinant soluble monomeric EphB4 exerts an inhibitory effect on choroidal angiogenesis in vitro and in vivo. It should be further evaluated for its potential as a novel therapy for CNV.