Localization of VEGF receptor-2 (KDR/Flk-1) and effects of blocking it in oxygen-induced retinopathy.
Localization of VEGF receptor-2 (KDR/Flk-1) and effects of blocking it in oxygen-induced retinopathy.
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发表时间：
2002-02
影响因子：
4.4
通讯作者：
D. Mcleod;M. Taomoto;Jingtai Cao;Zhen-ping Zhu;L. Witte;G. Lutty
中科院分区：
文献类型：
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作者：
D. Mcleod;M. Taomoto;Jingtai Cao;Zhen-ping Zhu;L. Witte;G. Lutty
血管内皮细胞生长因子（VEGF）与血管发育和增殖性视网膜病变有关。本研究的目的是检查免疫组化定位和相对水平的VEGF受体-2（KDR）在犬视网膜出生后血管生成和血管生成过程中的氧诱导视网膜病变（OIR），并调查这些过程中的中和KDR的影响。方法将1 ~ 22日龄正常犬眼和氧疗犬眼快速冷冻，用抗人KDR抗体进行免疫组化分析。为了检查阻断KDR的效果，将含有对照IgG或抗KDR的缓释聚合物丸通过手术植入6日龄空气饲养的对照和氧处理的动物。使用结合测定（测量与可溶性KDR的结合）评估从沉淀物洗脱的材料，以确定抗KDR释放的动力学和内皮细胞增殖，以测量生物活性。在22日龄时处死动物，用血管的腺苷二磷酸酶（ADP 13）组织化学染色检查组织。结果KDR免疫反应性与视网膜血管的发育仅弱相关，在正常的生后发育过程中，在成血管细胞中未观察到。免疫反应性是非常强的重组视网膜血管和玻璃体内新生血管在氧处理的动物。抗KDR对空气饲养的对照动物的血管形态或生长没有影响。在氧处理的动物中，与对照IgG颗粒眼相比，抗KDR显著抑制视网膜的血管再生（P = 0.005）和玻璃体内新生血管的形成（P < 0.04）。结论：KDR/Flk-1与正常视网膜血管的发育关系不明显，但在高氧损伤后视网膜血管新生和玻璃体内新生血管形成过程中的增殖内皮细胞中表达。抗KDR抗体缓释微丸对正常血管生成无影响，但可抑制OIR中玻璃体内新生血管的形成和视网膜血管的发育，提示阻断KDR可能有利于治疗成人组织病理性血管生成。
PURPOSE Vascular endothelial cell growth factor (VEGF) has been implicated in vascular development and in proliferative retinopathies. The goal of this study was to examine the immunohistochemical localization and relative levels of VEGF receptor-2 (KDR) in canine retina during postnatal vasculogenesis and during angiogenesis in oxygen-induced retinopathy (OIR) and to investigate the effects of neutralizing KDR on these processes. METHODS Eyes from normal dogs ranging from 1 to 22 days of age and age-matched oxygen-treated animals were snap frozen for immunohistochemical analysis with antibodies against human KDR. To examine the effects of blocking KDR, 6-day-old air-reared control and oxygen-treated animals were surgically implanted with slow release polymer pellets containing control IgG or anti-KDR. Material eluted from pellets was assessed using a binding assay (measures binding to soluble KDR) to determine the kinetics of anti-KDR release and endothelial cell proliferation to measure bioactivity. Animals were killed at 22 days of age and tissues examined with adenosine diphosphatase (ADPase) histochemical staining of blood vessels. RESULTS KDR immunoreactivity was only weakly associated with developing retinal vessels and was not observed in angioblasts throughout normal postnatal development. Immunoreactivity was very strong in reforming retinal vessels and intravitreal neovascularization in oxygen-treated animals. Anti-KDR had no effect on vessel morphology or growth in air-reared control animals. In oxygen-treated animals, anti-KDR significantly inhibited revascularization of the retina (P = 0.005) and formation of intravitreal neovascularization compared with control IgG pellet eyes (P < 0.04). CONCLUSIONS KDR/Flk-1 was only weakly associated with normal developing primary retinal vessels but was strongly expressed by proliferating endothelial cells in reforming retinal vessels and intravitreal neovascularization after hyperoxic insult. Anti-KDR antibody delivered by slow-release pellets had no effect on normal vasculogenesis, but it inhibited the formation of intravitreal neovascularization and retinal vessel development in OIR. The study suggests that blocking KDR may be beneficial for treating pathologic angiogenesis in adult tissue.