Soluble forms of EphrinB2 and EphB4 reduce retinal neovascularization in a model of proliferative retinopathy
Soluble forms of EphrinB2 and EphB4 reduce retinal neovascularization in a model of proliferative retinopathy
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DOI：
10.1167/iovs.04-0983
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发表时间：
2005-06-01
影响因子：
4.4
通讯作者：
Powers, MR
中科院分区：
文献类型：
--
作者：
Zamora, DO;Davies, MH;Powers, MR
目的. Ephrin配体及其Eph受体是哺乳动物血管发育过程中内皮细胞（EC）增殖、迁移、粘附和排斥的关键调节因子。假设这些分子也在氧诱导的视网膜病变小鼠模型中的病理性新生血管形成（NV）中发挥作用。出生后第7天（P）的C57 BL/6小鼠暴露于75%氧气（O-2）5天（直至P12），并允许在室内空气中恢复以诱导视网膜NV。通过RT-PCR特异性分析P7至P24之间来自未暴露和高氧暴露小鼠的视网膜的EphrinB 2和EphB 4转录物表达。磷酸化Eph（p-Eph）受体在活跃的EC增殖在P15和P17通过免疫组织学进行了评估。一些高氧暴露小鼠的一只眼睛在从高O-2过渡到室内空气（P12）期间玻璃体内注射150 ng/1.5 μ L可溶性EphrinB 2/Fc或EphB 4/Fc嵌合体，并在P14再次注射。将人IgG注射到侧眼作为对照。在疾病高峰期（P17）定量视网膜前核和视网膜血管。EphrinB 2 mRNA在发育中的视网膜中组成型表达，并且不受高氧的影响。相反，EphB 4 mRNA的表达在正常视网膜发育过程中受到调节，并被高氧改变。此外，在发育中的视网膜前簇中检测到p-Eph，因此暗示Ephrin/Eph信号传导系统在该实验模型中是活跃的。玻璃体内注射这些分子的可溶性版本显着减少病理性新血管形成。与对照注射相比，在EphrinB 2注射的眼睛中，高氧处理的小鼠中视网膜前核的数量减少了66%（P < 0.05），而EphB 4处理产生了69%的减少（P < 0.05）。视网膜内血管的发育没有被注射改变。这些结果支持内源性EphrinB 2和EphB 4是氧诱导的视网膜病变期间视网膜NV的调节剂并且可能是治疗干预的有用靶点的假设。
PURPOSE. Ephrin ligands and their Eph receptors are key regulators of endothelial cell (EC) proliferation, migration, adhesion, and repulsion during mammalian vascular development. The hypothesis was that these molecules also play a role in pathologic neovascularization (NV) in the mouse model of oxygen-induced retinopathy.METHODS. C57BL/6 mice at postnatal day (P) 7 were exposed to 75% oxygen (O-2) for 5 days (until P12) and allowed to recover in room air to induce retinal NV. Retinas from unexposed and hyperoxia-exposed mice between P7 to P24 were analyzed specifically for EphrinB2 and EphB4 transcript expression by RT-PCR. Phospho-Eph (p-Eph) receptor was evaluated during active EC proliferation at P15 and P17 by immunohistology. Some hyperoxia-exposed mice had one eye injected intravitreally with 150 ng/1.5 mu L of soluble EphrinB2/Fc or EphB4/Fc chimeras during transition from high O-2 to room air (P12) and injected again on P14. Contralateral eyes were injected with human IgG as the control. Preretinal nuclei and retinal blood vessels were quantified at peak disease (P17).RESULTS. EphrinB2 mRNA was constitutively expressed in the developing retina and was unchanged by hyperoxia. In contrast, EphB4 mRNA expression was modulated during normal retinal development and was altered by hyperoxia. Furthermore, p-Eph was detected in developing preretinal tufts, thus implying that Ephrin/Eph signaling system is active in this experimental model. Intravitreal injection of soluble versions of these molecules significantly reduced pathologic neovascularization. The number of preretinal nuclei in hyperoxia-treated mice was reduced by 66% (P < 0.05) in EphrinB2-injected eyes, whereas EphB4 treatment yielded a 69% reduction (P < 0.05), compared with control injections. Intraretinal vessel development was not altered by the injections.CONCLUSIONS. These results support the hypothesis that endogenous EphrinB2 and EphB4 are regulators of retinal NV during oxygen-induced retinopathy and may be useful targets for therapeutic intervention.