Vascular endothelial growth factor(VEGF)-A165-induced prostacyclin synthesis requires the activation of VEGF receptor-1 and-2 heterodimer
Vascular endothelial growth factor(VEGF)-A165-induced prostacyclin synthesis requires the activation of VEGF receptor-1 and-2 heterodimer
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DOI：
10.1074/jbc.m412017200
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发表时间：
2005-03-18
影响因子：
4.8
通讯作者：
Sirois, MG
中科院分区：
文献类型：
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作者：
Neagoe, PE;Lemieux, C;Sirois, MG
我们先前报道，血管内皮生长因子-A(165)的炎症作用是通过血管内皮细胞合成急性血小板激活因子，激活血管内皮生长因子受体-2(VEGFR-2)及其辅助受体神经粘连蛋白-1(NRP-1)来实现的。此外，血管内皮生长因子-A(165)促进其他内皮介质的释放，包括一氧化氮和前列环素(PGI(2))。然而，目前尚不清楚血管内皮生长因子-A(165)是否通过血管内皮生长因子受体-1(VEGFR-1)和/或血管内皮生长因子受体-2(VEGFR-2)的激活来调节PGI2的合成，以及辅助受体NRP-1是否增强了血管内皮生长因子-A(165)的活性。本研究采用酶联免疫吸附试验对牛主动脉内皮细胞(BAEC)的稳定代谢产物6-酮-前列腺素F-1α(6-keto-PGF)、6-酮-前列腺素F_(1α)(6-keto-PGF(1α))进行定量研究。用血管内皮生长因子类似物、血管内皮生长因子A(165)(VEGFR-1、VEGFR-2和NRP-1激动剂)和血管内皮生长因子-A(121)(VEGFR-1和VEGFR-2激动剂)(高达10(-9)M)处理BAEC，在15分钟内使PGI2合成分别增加70倍和40倍。用VEGFR-1(胎盘生长因子和血管内皮生长因子-B)或VEGFR-2(血管内皮生长因子-C)激动剂治疗不能增加PGI2的合成。VEGFR-1和VEGFR-2激动剂联合应用不能增加PGI2的释放。VEGFR-2抑制剂预处理可抑制由VEGFR-1和VEGFR-2mRNA介导的PGI2释放，靶向VEGFR-1或VEGFR-2mRNA的反义寡聚物可使血管内皮细胞由VEGFR-1和VEGFR-2mRNA介导的PGI2合成减少高达79%。综上所述，我们的研究结果表明，VEGFR-1和VEGFR-2异源二聚体(VEGFR-1/R-2)的激活是由VEGFR-A(165)和VEGFR-A(121)介导的PGI2合成所必需的，而VEGFR-1和VEGFR-2同源二聚体与相应激动剂的平行激活不能复制这种作用。此外，VEGF-A(165)与NRP-1的结合增强了其促进PGI(2)合成的能力。
We previously reported that vascular endothelial growth factor (VEGF)-A(165) inflammatory effect is mediated by acute platelet-activating factor synthesis from endothelial cells upon the activation of VEGF receptor-2 (VEGFR-2) and its coreceptor, neuropilin-1 (NRP-1). In addition, VEGF-A(165) promotes the release of other endothelial mediators including nitric oxide and prostacyclin (PGI(2)). However, it is unknown whether VEGF-A(165) is mediating PGI2 synthesis through VEGF receptor-1 (VEGFR-1) and/or VEGF receptor-2 (VEGFR-2) activation and whether the coreceptor NRP-1 potentiates VEGF-A(165) activity. In this study, PGI2 synthesis in bovine aortic endothelial cells (BAEC) was assessed by quantifying its stable metabolite (6- keto prostaglandin F-1 alpha, 6- keto PGF(1 alpha)) by enzyme-linked immunosorbent assay. Treatment of BAEC with VEGF analogs, VEGF-A(165) (VEGFR-1, VEGFR-2 and NRP-1 agonist) and VEGF-A(121) ( VEGFR-1 and VEGFR-2 agonist) ( up to 10(-9) M), increased PGI2 synthesis by 70- and 40-fold within 15 min. Treatment with VEGFR-1 ( placental growth factor and VEGF-B) or VEGFR-2 (VEGF-C) agonist did not increase PGI2 synthesis. The combination of VEGFR-1 and VEGFR-2 agonists did not increase PGI2 release. Pretreatment with a VEGFR-2 inhibitor abrogated PGI2 release mediated by VEGF-A(165) and VEGF-A(121), and pretreatment of BAEC with antisense oligomers targeting VEGFR-1 or VEGFR-2 mRNA reduced PGI2 synthesis mediated by VEGF-A(165) and VEGF-A(121) up to 79%. In summary, our data demonstrate that the activation of VEGFR-1 and VEGFR-2 heterodimer (VEGFR-1/R-2) is essential for PGI2 synthesis mediated by VEGF-A(165) and VEGF-A(121), which cannot be reproduced by the parallel activation of VEGFR-1 and VEGFR-2 homodimers with corresponding agonists. In addition, the binding of VEGF-A(165) to NRP-1 potentiates its capacity to promote PGI(2) synthesis.