Low molecular weight fucoidan increases VEGF165-induced endothelial cell migration by enhancing VEGF165 binding to VEGFR-2 and NRP1
Low molecular weight fucoidan increases VEGF165-induced endothelial cell migration by enhancing VEGF165 binding to VEGFR-2 and NRP1
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DOI：
10.1074/jbc.m600686200
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发表时间：
2006-12-08
影响因子：
4.8
通讯作者：
Letourneur, Didier
中科院分区：
文献类型：
--
作者：
Lake, Andrew C.;Vassy, Roger;Letourneur, Didier
治疗性诱导血管生成是治疗慢性缺血的一种潜在方法。硫酸乙酰肝素蛋白多糖通过与促血管生成因子如血管内皮生长因子(VEGF)的相互作用而发挥重要作用。低分子褐藻糖胶(LMWF)是从棕色海藻中提取的一种模拟肝素某些生物活性的硫酸化多糖，最近被证明能促进大鼠严重的后肢缺血后的血管重建。在这份报告中，我们首先使用培养的人内皮细胞(ECs)来研究LMWF可能增强血管内皮生长因子(165)的作用。数据显示，LMWF在生长因子降低的基质中极大地促进EC管的形成。LMWF对VEGF(165)诱导的EC趋化作用有很强的促进作用，但不能促进EC的增殖。此外，LMWF对血管内皮生长因子(121)诱导的EC迁移没有影响，内皮细胞迁移是一种不与硫酸乙酰肝素蛋白多糖结合的血管内皮生长因子亚型。然后，通过I-125-VEGF(165)的结合研究，我们观察到LMWF增强了VEGF(165)与重组VEGFR-2和Neuropilin-1(Nrp1)的结合，但不增强与VEGFR-1的结合。表面等离子体共振分析表明，LMWF与血管内皮生长因子165(1.2 nM)及其受体(5-20 nM)有较高的亲和力，但不与血管内皮生长因子121结合。预先在固定化受体上注射LMWF表明，与VEGFR-1相比，VEGF(165)对VEGFR-2和Nrp1具有最高的亲和力。总体而言，低分子肝素的作用比低分子肝素明显得多。提示LMWF通过促进血管内皮细胞上VEGFR-2和Nrp1与VEGFR-2和Nrp1的结合，从而促进治疗性血管重建，从而发挥有效的作用机制。
Therapeutic induction of angiogenesis is a potential treatment for chronic ischemia. Heparan sulfate proteoglycans are known to play an important role by their interactions with proangiogenic growth factors such as vascular endothelial growth factor (VEGF). Low molecular weight fucoidan (LMWF), a sulfated polysaccharide from brown seaweeds that mimic some biological activities of heparin, has been shown recently to promote revascularization in rat critical hindlimb ischemia. In this report, we first used cultured human endothelial cells (ECs) to investigate the possible ability of LMWF to enhance the actions of VEGF(165). Data showed that LMWF greatly enhances EC tube formation in growth factor reduced matrigel. LMWF is a strong enhancer of VEGF(165)-induced EC chemotaxis, but not proliferation. In addition, LMWF has no effect on VEGF(121)-induced EC migration, a VEGF isoform that does not bind to heparan sulfate proteoglycans. Then, with binding studies using I-125-VEGF(165), we observed that LMWF enhances the binding of VEGF(165) to recombinant VEGFR-2 and Neuropilin-1 (NRP1), but not to VEGFR-1. Surface plasmon resonance analysis showed that LMWF binds with high affinity to VEGF(165) (1.2 nM) and its receptors (5-20 nM), but not to VEGF(121). Pre-injection of LMWF on immobilized receptors shows that VEGF(165) has the highest affinity for VEGFR-2 and NRP1, as compared with VEGFR-1. Overall, the effects of LMWF were much more pronounced than those of LMW heparin. These findings suggested an efficient mechanism of action of LMWF by promoting VEGF(165) binding to VEGFR- 2 and NRP1 on ECs that could help in stimulating therapeutic revascularization.