Thrombin receptor-mediated increase of two matrix metalloproteinases, MMP-1 and MMP-3, in human endothelial cells
Thrombin receptor-mediated increase of two matrix metalloproteinases, MMP-1 and MMP-3, in human endothelial cells
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DOI：
10.1161/01.atv.17.10.1931
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发表时间：
1997-10-01
影响因子：
8.7
通讯作者：
KleinSoyer, C
中科院分区：
文献类型：
--
作者：
DuhamelClerin, E;Orvain, C;KleinSoyer, C
基质金属蛋白酶（MMPs）负责细胞外基质成分的降解，由包括人内皮细胞在内的多种细胞分泌。由于已知α -凝血酶与基质成分相互作用，并已被证明能激活人脐静脉内皮细胞中潜伏的MMP-2，我们研究了人α -凝血酶是否也能调节人隐静脉或乳腺动脉内皮细胞（EC）分泌的其他MMPs。在不同时期增加凝血酶浓度处理EC后，除了MMP-2激活外，还观察到MMP-1和MMP-3的明胶溶解活性显著提高。凝血酶的作用是时间和剂量依赖性的，在24小时达到最大。5 NIH U/ml凝血酶处理24小时后，Western blotting显示MMP-3和MMP-1分别比对照组增加9.5倍和4.4倍。合成的凝血酶受体激动剂肽SFLLRNPNDKYEPF完全复制了凝血酶的作用，而凝血酶催化位点的化学失活则消除了其对MMP-1和MMP-3的作用。凝血酶和SFLLRNPNDKYEPF均诱导MMP-3 mRNA合成，但对组成型MMP-1 mRNA水平无显著影响。这些结果表明凝血酶不仅激活潜伏的MMP-2，而且还调节EC中MMP-1和MMP-3的产生，后者的作用是由g蛋白偶联凝血酶受体介导的。因此，我们目前的数据为支持凝血酶在组织重塑和血管生成中的作用提供了证据。
Matrix metalloproteinases (MMPs) are responsible for the degradation of extracellular matrix components and are secreted by a variety of cells including human endothelial cells. Because alpha-thrombin is known to interact with matrix components and has been shown to activate latent MMP-2 in human umbilical vein endothelial cells, we investigated whether human alpha-thrombin could also regulate other MMPs secreted by the human saphenous vein or mammary artery endothelial cells (EC). After treatment of EC with increasing concentrations of thrombin for different periods of time, a significantly higher gelatinolytic activity of both MMP-1 and MMP-3 was observed in addition to MMP-2 activation. The effect of thrombin was time and dose-dependent, reaching a maximum at 24 hours. After treatment with 5 NIH U/ml thrombin for 24 hours, Western blotting revealed 9.5- and 4.4-fold increases over control values for MMP-3 and MMP-1, respectively. The synthetic thrombin receptor agonist peptide SFLLRNPNDKYEPF fully reproduced the action of thrombin, whereas chemical inactivation of the catalytic site of thrombin abolished its effect on MMP-1 and MMP-3. Thrombin and SFLLRNPNDKYEPF both induced MMP-3 mRNA synthesis but had no significant influence on constitutive MMP-1 mRNA levels. These results demonstrate that thrombin not only activates latent MMP-2 but also modulates MMP-1 and MMP-3 production in EC, this latter effect being mediated by the G-protein-coupled thrombin receptor. Hence, our present data provide evidence to support the suspected role of thrombin in tissue remodeling and angiogenesis.