Nerve growth factor-induced migration of endothelial cells
Nerve growth factor-induced migration of endothelial cells
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DOI：
10.1124/jpet.105.093252
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发表时间：
2005-12-01
影响因子：
3.5
通讯作者：
Lelkes, PI
中科院分区：
文献类型：
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作者：
Dollé, JP;Rezvan, A;Lelkes, PI
神经生长因子（NGF）是神经系统中众所周知的神经营养性和神经营养性激动剂，最近被证明还能诱导内皮细胞（EC）的血管生成作用。为了测量 NGF 对培养的 EC 迁移（新生血管生成的重要步骤）的影响，我们优化了使用人主动脉内皮细胞 (HAEC) 的全向迁移测定，并使用人重组碱性成纤维细胞生长因子 (rhbFGF) 和人重组血管内皮生长因子 (rhVEGF) 验证了该测定。从不同物种（蝰蛇、小鼠和重组人）纯化的神经生长因子刺激 HAEC 迁移的效力与 VEGF 和碱性成纤维细胞生长因子 (bFGF) 相似（EC50 与 0.5 ng/ml 相似）。重组人 bFGF 明显比 viper NGF 或 rhVEGF 更有效，这两种药物都能刺激 HAEC 迁移，比基础自发迁移提高约 30%。 NGF 介导的 HAEC 迁移刺激被 NGF/TrkA 受体拮抗剂 K252a 完全阻断 [(8R*, 9S*, 11S*)-(i)-9-羟基-9-甲氧基羰基-8-甲基-2,3,9,10-四氢- 8,11-环氧-1H,8H, 11H-2,7b, 11a-三氮杂二苯并(a,g) cycloocta(c,d,e) trindene-1one] (30 nM)，但不是 VEGF/Flk 受体拮抗剂 SU-5416 [3-[(2,4-二甲基吡咯-5-基)亚甲基]-indolin-2one] (250 nM)，表明 NGF 通过 TrkA 受体激活对 HAEC 迁移产生直接影响。 rhVEGF 或 rhbFGF 进一步增强 Viper NGF 对 HAEC 迁移的刺激，表明它们的酪氨酸激酶受体信号通路之间存在增强的相互作用。 Viper NGF 代表了一种新的药理学工具，用于研究内皮细胞中可能的 TrkA 受体亚型。 NGF在体外刺激HAEC细胞迁移的能力意味着该因子除了其在神经系统中众所周知的作用之外，可能在心血管系统中发挥重要作用。
Nerve growth factor (NGF) is a well known neurotropic and neurotrophic agonist in the nervous system, which recently was shown to also induce angiogenic effects in endothelial cells (ECs). To measure NGF effects on the migration of cultured ECs, an important step in neoangiogenesis, we optimized an omnidirectional migration assay using human aortic endothelial cells (HAECs) and validated the assay with human recombinant basic fibroblast growth factor (rhbFGF) and human recombinant vascular endothelial growth factor (rhVEGF). The potencies of nerve growth factor purified from various species ( viper, mouse, and recombinant human) to stimulate HAEC migration was similar to that of VEGF and basic fibroblast growth factor (bFGF) (EC50 of similar to 0.5 ng/ml). Recombinant human bFGF was significantly more efficacious than either viper NGF or rhVEGF, both of which stimulated HAEC migration by similar to 30% over basal spontaneous migration. NGF-mediated stimulation of HAEC migration was completely blocked by the NGF/TrkA receptor antagonist K252a [(8R*, 9S*, 11S*)-(i)-9-hydroxy-9-methoxycarbonyl-8-methyl-2,3,9,10-tetrahydro- 8,11-epoxy-1H,8H, 11H-2,7b, 11a-triazadibenzo(a,g) cycloocta(c,d,e) trindene-1one] (30 nM) but not by the VEGF/Flk receptor antagonist SU-5416 [3-[(2,4-dimethylpyrrol-5-yl) methylidenyl]-indolin-2one] (250 nM), indicating a direct effect of NGF via TrkA receptor activation on HAEC migration. Viper NGF stimulation of HAEC migration was additively increased by either rhVEGF or rhbFGF, suggesting a potentiating interaction between their tyrosine kinase receptor signaling pathways. Viper NGF represents a novel pharmacological tool to investigate possible TrkA receptor subtypes in endothelial cells. The ability of NGF to stimulate migration of HAEC cells in vitro implies that this factor may play an important role in the cardiovascular system besides its well known effects in the nervous system.