IP-10 blocks vascular endothelial growth factor-induced endothelial cell motility and tube formation via inhibition of calpain
IP-10 blocks vascular endothelial growth factor-induced endothelial cell motility and tube formation via inhibition of calpain
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DOI：
10.1161/01.res.0000209968.66606.10
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发表时间：
2006-03-17
影响因子：
20.1
通讯作者：
Wells, A
中科院分区：
文献类型：
--
作者：
Bodnar, RJ;Yates, CC;Wells, A
血管生成在创伤修复中起着关键作用。内皮细胞呈递CXC受体3（CXCR 3），用于在伤口再生后期表达的趋化因子。为了了解CXCR 3在调节内皮功能中的生理作用，我们分析了CXCR 3配体IP-10（CXCL 10）影响内皮细胞管形成的能力。在血管内皮生长因子（VEGF）存在下用IP-10处理内皮细胞抑制生长因子减少的基质胶上和皮下基质胶塞中的管形成。此外，IP-10显着抑制VEGF诱导的内皮运动，血管生成的关键反应。以前的工作表明，CXCR 3配体启动蛋白激酶A（PKA）磷酸化依赖性抑制m-钙蛋白酶，诱导细胞运动所需的，在成纤维细胞，但不是上皮细胞。在这里，我们表明内皮细胞中的CXCR 3激活会诱导cAMP和PKA激活的增加。用PKA抑制剂Rp-8-Br-cAMP或PKA的小干扰RNA处理内皮细胞能够逆转IP-10对VEGF介导的管形成和运动的抑制作用。重要的是，用VEGF处理内皮细胞诱导了m-钙蛋白酶的激活，但与IP-10的共刺激显著降低了这种活性。使用Rp-8-Br-cAMP，我们显示阻断PKA逆转了IP-10对VEGF诱导的m-钙蛋白酶活性的抑制。这些数据表明CXCR 3的激活通过PKA介导的m-钙蛋白酶抑制抑制内皮管形成。这提供了一种手段，通过该手段，晚期伤口修复信号限制在伤口响应过程早期驱动的血管生成。
Angiogenesis plays a critical role in wound repair. Endothelial cells present CXC receptor 3 (CXCR3) for chemokines expressed late in wound regeneration. To understand the physiological role CXCR3 plays in regulating endothelial function, we analyzed the ability of a CXCR3 ligand, IP-10 (CXCL10), to influence endothelial cell tube formation. Treatment of endothelial cells with IP-10 in the presence of vascular endothelial growth factor (VEGF) inhibited tube formation on growth factor-reduced Matrigel and in a subcutaneous Matrigel plug. Furthermore, IP-10 significantly inhibited VEGF-induced endothelial motility, a response critical for angiogenesis. Previous work showed that CXCR3 ligandation initiates protein kinase A (PKA) phosphorylation-dependent inhibition of m-calpain, required for induced cell motility, in fibroblasts but not epithelial cells. Here we show that CXCR3 activation in endothelial cells induces an increase in cAMP and PKA activation. Treatment of endothelial cells with Rp-8-Br-cAMP, an inhibitor of PKA, or small interference RNA to PKA was able to reverse the inhibitory effects of IP-10 on VEGF-mediated tube formation and motility. Importantly, treatment of endothelial cells with VEGF induced the activation of m-calpain, but costimulation with IP-10 significantly decreased this activity. Using Rp-8-Br-cAMP, we show blocking PKA reversed the IP-10 inhibition of VEGF-induced m-calpain activity. These data indicate that the activation of CXCR3 inhibits endothelial tube formation through a PKA mediated inhibition of m-calpain. This provides a means by which late wound repair signals limit the angiogenesis driven early in the wound response process.