Sakamoto Takayuki, Ishibashi Toshiyuki, Sakamoto Nobuo, Sugimoto Koichi, Egashira Kensuke, Ohkawara Hiroshi, Nagata Kenji, Yokoyama Keiko, Kamioka Masashi, Ichiki Toshihiro, Sugimoto Naotoshi, Kurabayashi Masahiko, Suzuki Koji, Takuwa Yoh, Maruyama Yukio
First Department of Internal Medicine, Fukushima Medical University, 1 Hikarigaoka, Fukushima, 960-1295, Japan.
Arterioscler Thromb Vasc Biol. 2005 Sep;25(9):2005-11. doi: 10.1161/01.ATV.0000178171.61754.cd. Epub 2005 Jul 14.
Ca2+ plays an important role in tissue factor (TF) gene expression. We investigated the role of endogenous nitric oxide (NO) in the induction of TF expression in endothelial cells (ECs) by monocyte adhesion and the mechanisms of NO action.
Inhibition of endogenous NO by Nomega-nitro-L-arginine methyl ester (L-NAME) enhanced TF promoter activity and protein expression induced in human coronary ECs by monocyte adhesion, as well as EC surface TF activity. L-NAME also induced monocyte chemoattractant protein-1 (MCP-1) expression, which was blocked by an NO donor, NOC18. Exogenous MCP-1 enhanced TF expression induced by monocyte adhesion, whereas adenovirus-mediated expression of the mutant MCP-1, 7ND, abolished the L-NAME enhancement of TF expression induced by monocyte adhesion. Monocyte attachment to L-NAME-treated ECs increased Ca2+ influx, which was prevented by NOC18, anti-MCP-1 antibody or 7ND. These results indicate that the binding of increased MCP-1 induced by endogenous NO blockade to CCR2 mediated the enhancement of Ca2+ influx only when monocytes adhered to ECs, which upregulated TF expression in ECs triggered by monocyte adhesion.
MCP-1/CCR2 may play a role in Ca2+ influx-dependent TF regulation in the monocyte-EC interaction in the impairment of NO synthesis.
钙离子在组织因子（TF）基因表达中起重要作用。我们研究了内源性一氧化氮（NO）在单核细胞黏附诱导内皮细胞（ECs）中TF表达的作用以及NO作用的机制。
用Nω-硝基-L-精氨酸甲酯（L-NAME）抑制内源性NO可增强单核细胞黏附诱导的人冠状动脉ECs中TF启动子活性和蛋白表达，以及EC表面TF活性。L-NAME还诱导单核细胞趋化蛋白-1（MCP-1）表达，而这被NO供体NOC18阻断。外源性MCP-1增强了单核细胞黏附诱导的TF表达，而腺病毒介导的突变型MCP-1（7ND）的表达消除了L-NAME对单核细胞黏附诱导的TF表达的增强作用。单核细胞与L-NAME处理的ECs的附着增加了钙离子内流，这被NOC18、抗MCP-1抗体或7ND阻止。这些结果表明，内源性NO阻断诱导的MCP-1增加与CCR2的结合仅在单核细胞黏附于ECs时介导了钙离子内流的增强，从而上调了单核细胞黏附触发的ECs中TF的表达。
MCP-1/CCR2可能在NO合成受损时单核细胞与EC相互作用中钙离子内流依赖性TF调节中起作用。
