Role of the JNK pathway in thrombin-induced ICAM-1 expression in endothelial cells
Role of the JNK pathway in thrombin-induced ICAM-1 expression in endothelial cells
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DOI：
10.1016/j.cardiores.2005.05.029
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发表时间：
2005-11-01
影响因子：
10.8
通讯作者：
Chayama, K
中科院分区：
文献类型：
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作者：
Miho, N;Ishida, T;Chayama, K
目的：凝血酶通过增加细胞间粘附分子-1（ICAM-1）的表达诱导白细胞粘附内皮细胞。尽管ICAM-1的表达受NF-κ B B的调节，但最近的研究表明，可能还涉及其他信号传导机制。本研究的目的是确定丝裂原活化蛋白（MAP）激酶，包括细胞外信号调节激酶1/2（ERK 1/2）、c-Jun N末端激酶（JNK）和p38 MAP激酶（p38）是否介导凝血酶诱导的内皮细胞ICAM-1表达。使用抗ICAM-1的蛋白质印迹分析1抗体和荧光素酶测定在加入信号转导抑制剂或转染各种基因构建体。通过使用c-Jun作为底物的激酶测定或通过使用抗磷酸化JNK抗体的蛋白质印迹分析来测定JNK激酶活性。用JNK特异性抑制剂SP 600125或JNK抑制肽1（JNKI 1）处理内皮细胞，结果凝血酶诱导的ICAM-1表达显著降低，如Western印迹分析所示（分别为67 +/-3%和72 +/-7%）。相比之下，MEK和p38的抑制剂仅具有最小的效果。SP 600125和NF-κ B抑制剂BAY 11-7082的组合导致凝血酶诱导的ICAM-1表达的完全抑制。G（alpha q）抑制剂YM-254890抑制凝血酶诱导的JNK活化和ICAM-1表达。显性负性Ras和Rac 1，但不Rho，抑制凝血酶诱导的JNK激活和ICAM-1启动子活性。最后，凝血酶诱导的JNK激活和ICAM-1启动子活性被抑制β ARK 1ct（G β γ亚基清除剂）和Csk.Conclusions：这些数据表明，在音乐会与NF-κ B B，JNK调节凝血酶诱导的ICAM-1表达的机制是依赖于G（α q），G β γ，Ras，Rac 1和Src激酶家族。(c)2005年欧洲心脏病学会。Elsevier B. V.出版，保留所有权利。
Objective: Thrombin induces leukocyte adherence to endothelial cells via increased expression of intercellular adhesion molecule-1 (ICAM-1). Although ICAM-1 expression is regulated by NF-kappa B, recent studies have suggested that additional signaling mechanisms may also be involved. The goal of this study was to determine whether mitogen-activated protein (MAP) kinases, including extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38 MAP kinase (p38), mediate thrombin-induced ICAM-1 expression in endothelial cells.Methods: Western blot analysis using anti-ICAM-1 antibody and luciferase assays were performed in cultured endothelial cells after addition of signal transduction inhibitors or transfection of various gene constructs. JNK kinase activity was determined by a kinase assay using c-Jun as a substrate or by Western blot analysis with anti-phospho-JNK antibody.Results: Treatment of endothelial cells with the JNK-specific inhibitors, SP600125 or JNK inhibitory peptide 1 (JNKI1), resulted in a significant decrease in thrombin-induced ICAM-1 expression as demonstrated by Western blot analysis (67 +/- 3% and 72 +/- 7%, respectively). In contrast, inhibitors of MEK and p38 had only minimal effect. The combination of SP600125 and the NF-kappa B inhibitor, BAY 11-7082, resulted in complete inhibition of thrombin-induced ICAM-1 expression. The G(alpha q) inhibitor, YM-254890, inhibited thrombin-induced JNK activation and ICAM-1 expression. Dominant-negative Ras and Rac1, but not Rho, inhibited thrombin-induced JNK activation and ICAM-1 promoter activity. Finally, thrombin-induced JNK activation and ICAM-1 promoter activity were inhibited by beta ARK1ct (a G beta gamma subunit scavenger) and Csk.Conclusions: These data suggest that, in concert with NF-kappa B, JNK regulates thrombin-induced ICAM-1 expression by a mechanism that is dependent on G(alpha q), G beta gamma, Ras, Rac1 and the Src kinase family. (c) 2005 European Society of Cardiology. Published by Elsevier B.V. All rights reserved.