Src kinase mediates angiotensin II-dependent increase in pulmonary endothelial nitric oxide synthase.
Src kinase mediates angiotensin II-dependent increase in pulmonary endothelial nitric oxide synthase.
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DOI：
10.1165/rcmb.2004-0098oc
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发表时间：
2004-06
影响因子：
6.4
通讯作者：
Xinmei Li;K. Lerea;Jianyu Li;S. Olson
中科院分区：
文献类型：
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作者：
Xinmei Li;K. Lerea;Jianyu Li;S. Olson
我们以前已经证明，血管紧张素II（Ang II）刺激一氧化氮（NO）的生产在牛肺动脉内皮细胞（BPAECs）通过增加NO合成酶（NOS）的表达，通过2型受体。本研究的目的是确定血管紧张素II依赖的信号通路，介导这种增加内皮型一氧化氮合酶（eNOS）。当用酪氨酸激酶抑制剂除莠霉素A和4-氨基-5-（4-氯苯基）-7-（叔丁基）吡唑并[3，4-D]嘧啶预处理BPAEC时，可阻止Ang II依赖的eNOS表达增加，这也阻断了Ang II依赖的丝裂原活化蛋白激酶（MAPK）激酶/细胞外调节蛋白激酶（MEK）-1和MAPK磷酸化，表明Src在该途径中位于MAPK的上游。用Src显性负突变cDNA转染BPAECs可阻止Ang II依赖的Src激活和eNOS蛋白表达的增加。MEK-1抑制剂PD 98059可抑制细胞外调节蛋白激酶1和2的Ang II依赖性磷酸化，并抑制eNOS表达的增加。无论是AG 1478，表皮生长因子受体激酶抑制剂，也不是AG 1295，血小板衍生的生长因子受体激酶抑制剂，对血管紧张素II刺激的Src活性，MAPK激活，或eNOS表达有任何影响。百日咳毒素阻止了Src活性、MAPK激活和eNOS表达的Ang II依赖性增加。这些数据表明，血管紧张素II刺激Src酪氨酸激酶通过百日咳毒素敏感的途径，这反过来又激活MAPK途径，导致增加eNOS蛋白表达的BPAEC。
We have previously demonstrated that angiotensin II (Ang II) stimulates nitric oxide (NO) production in bovine pulmonary artery endothelial cells (BPAECs) by increasing NO synthase (NOS) expression via the type 2 receptor. The purpose of this study was to identify the Ang II-dependent signaling pathway that mediates this increase in endothelial NOS (eNOS). The Ang II-dependent increase in eNOS expression is prevented when BPAECs are pretreated with the tyrosine kinase inhibitors, herbimycin A and 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-D]pyrimidine, which also blocked Ang II-dependent mitogen-activated protein kinase (MAPK) kinase/extracellular-regulated protein kinase (MEK)-1 and MAPK phosphorylation, suggesting that Src is upstream of MAPK in this pathway. Transfection of BPAECs with an Src dominant negative mutant cDNA prevented the Ang II-dependent Src activation and increase in eNOS protein expression. PD98059, a MEK-1 inhibitor, prevented the Ang II-dependent phosphorylation of extracellular-regulated protein kinases 1 and 2 and increase in eNOS expression. Neither AG1478, an epidermal growth factor receptor kinase inhibitor, nor AG1295, a platelet derived growth factor receptor kinase inhibitor, had any effect on Ang II-stimulated Src activity, MAPK activation, or eNOS expression. Pertussis toxin prevented the Ang II-dependent increase in Src activity, MAPK activation, and eNOS expression. These data suggest that Ang II stimulates Src tyrosine kinase via a pertussis toxin-sensitive pathway, which in turn activates the MAPK pathway, resulting in increased eNOS protein expression in BPAECs.