Mechanisms of MAPK activation by bradykinin in vascular smooth muscle cells.
Mechanisms of MAPK activation by bradykinin in vascular smooth muscle cells.
复制标题
血管平滑肌细胞中缓激肽激活 MAPK 的机制。
DOI：
10.1152/ajpcell.1999.277.2.c253
复制
发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Jaffa,AA
中科院分区：
文献类型：
--
作者：
Velarde,V;Ullian,ME;Morinelli,TA;Mayfield,RK;Jaffa,AA
血管平滑肌细胞（VSMC）增殖是内皮损伤后发生的动脉粥样硬化过程的显著特征。已经描述了血管壁激肽释放酶-激肽系统。该系统对血管疾病的作用尚不明确。在本研究中，我们的特点的信号转导途径，导致丝裂原活化蛋白激酶（MAPK）激活的反应缓激肽（BK）在VSMC。向VSMC中添加10 - 10-10 - 7 M BK导致几种144至40 kDa蛋白质的酪氨酸磷酸化迅速且呈浓度依赖性增加。BK的这种作用可被B2-激肽受体拮抗剂HOE-140消除，但不能被B1-激肽受体拮抗剂des-Arg 9-Leu 8-BK消除。用抗磷酸酪氨酸抗体进行免疫沉淀，然后进行免疫印迹，结果显示10− 9 M BK诱导粘着斑激酶（p125 FAK）的酪氨酸磷酸化。BK（10− 8 M）促进p60 src与衔接蛋白生长因子受体结合蛋白-2的结合，也诱导MAPK活性显著增加。百日咳和霍乱毒素不抑制BK诱导的MAPK酪氨酸磷酸化。蛋白激酶C下调佛波酯12-肉豆蔻酸酯13-乙酸酯和/或抑制剂蛋白激酶C，p60 src激酶，MAPK激酶抑制BK诱导的MAPK酪氨酸磷酸化。这些发现提供了证据表明，VSMC中B2-激肽受体的激活导致多种第二信使的产生，这些第二信使聚集以激活MAPK。BK对这种关键激酶的激活为研究BK在血管损伤疾病状态下对VSMC增殖的促有丝分裂作用提供了强有力的理论基础。
Vascular smooth muscle cell (VSMC) proliferation is a prominent feature of the atherosclerotic process occurring after endothelial injury. A vascular wall kallikrein-kinin system has been described. The contribution of this system to vascular disease is undefined. In the present study we characterized the signal transduction pathway leading to mitogen-activated protein kinase (MAPK) activation in response to bradykinin (BK) in VSMC. Addition of 10−10–10−7M BK to VSMC resulted in a rapid and concentration-dependent increase in tyrosine phosphorylation of several 144- to 40-kDa proteins. This effect of BK was abolished by the B2-kinin receptor antagonist HOE-140, but not by the B1-kinin receptor antagonist des-Arg9-Leu8-BK. Immunoprecipitation with anti-phosphotyrosine antibodies followed by immunoblot revealed that 10−9M BK induced tyrosine phosphorylation of focal adhesion kinase (p125FAK). BK (10−8M) promoted the association of p60srcwith the adapter protein growth factor receptor binding protein-2 and also induced a significant increase in MAPK activity. Pertussis and cholera toxins did not inhibit BK-induced MAPK tyrosine phosphorylation. Protein kinase C downregulation by phorbol 12-myristate 13-acetate and/or inhibitors to protein kinase C, p60srckinase, and MAPK kinase inhibited BK-induced MAPK tyrosine phosphorylation. These findings provide evidence that activation of the B2-kinin receptor in VSMC leads to generation of multiple second messengers that converge to activate MAPK. The activation of this crucial kinase by BK provides a strong rationale to investigate the mitogenic actions of BK on VSMC proliferation in disease states of vascular injury.