Rho/Rho-kinase pathway contributes to C-reactive protein-induced plasminogen activator inhibitor-1 expression in endothelial cells
Rho/Rho-kinase pathway contributes to C-reactive protein-induced plasminogen activator inhibitor-1 expression in endothelial cells
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DOI：
10.1161/01.atv.0000183607.50230.9f
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发表时间：
2005-10-01
影响因子：
8.7
通讯作者：
Nakano, T
中科院分区：
文献类型：
--
作者：
Nakakuki, T;Ito, M;Nakano, T
目的- Rho/Rho激酶通路在动脉硬化和高血压等心血管疾病中起着关键作用。近年来，C反应蛋白（CRP）作为心血管事件的有力标志物，对血管细胞具有直接的促动脉粥样硬化作用已变得很明显。然而，其分子机制尚未得到充分研究。我们研究了Rho/Rho激酶信号通路参与CRP诱导的牛主动脉内皮细胞（BAECs）中纤溶酶原激活物抑制剂-1（派-1）表达的情况。RhoA活化通过使用rhotekin的Rho结合片段的亲和力下拉测定来确定。使用荧光素酶报告基因测定NF-κ B活性。BAEC与人重组CRP（>= 25 μ g/mL）孵育诱导派-1表达显著增加。用CRP刺激BAEC显著增加RhoA活化。TAT-C3（一种膜渗透性RhoA抑制剂）和Y-27632（Rho激酶抑制剂）预处理显著抑制CRP诱导的派-1表达。CRP可显著增强NF-κ B B的活性，而Y- 27632预处理可抑制其活化。Parthenoptera，SN 50，和BAY 11-7082（NF-κ B抑制剂）显着阻断CRP介导的派-1 expression.Conclusions -这些数据表明，CRP激活Rho/Rho激酶信号，这反过来又激活NF-κ B活性，导致派-1在BAEC中的表达。这些观察结果为Rho/Rho激酶信号转导可能参与CRP诱导的动脉粥样硬化血栓形成提供了证据。
Objective - Rho/Rho-kinase pathway plays pivotal roles in cardiovascular diseases including arteriosclerosis and hypertension. Recently it has become evident that C-reactive protein (CRP), a powerful marker for cardiovascular events, has direct proatherothrombotic effects on vascular cells. However, its molecular mechanism has not been fully investigated. We examined the involvement of Rho/Rho-kinase signaling in CRP-induced plasminogen activator inhibitor-1 (PAI-1) expression in bovine aortic endothelial cells (BAECs).Methods and Results - PAI-1 expression was determined by Western blotting. RhoA activation was determined by an affinity pull-down assay using Rho-binding fragment of rhotekin. NF-kappa B activity was determined using the luciferase reporter gene. Incubation of BAECs with human recombinant CRP ( >= 25 mu g/mL) induced a significant increase in PAI-1 expression. Stimulation of BAECs with CRP significantly increased RhoA activation. Pretreatment with TAT-C3 ( a membrane-permeable RhoA inhibitor) and Y-27632 ( Rho-kinase inhibitor) significantly inhibited CRP-induced PAI-1 expression. NF-kappa B activity was markedly enhanced by CRP and pretreatment with Y- 27632 inhibited its activation. Parthenolide, SN50, and BAY 11-7082 (NF-kappa B inhibitors) significantly blocked CRP-mediated PAI-1 expression.Conclusions - These data suggested that CRP activates Rho/Rho-kinase signaling, which in turn activates NF-kappa B activity, resulting in PAI-1 expression in BAEC. These observations provide evidence for the possible involvement of Rho/Rho-kinase signaling in CRP-induced atherothrombogenesis.