Involvement of Ca2+/calmodulin-dependent protein kinase II in endothelial NO production and endothelium-dependent relaxation
Involvement of Ca2+/calmodulin-dependent protein kinase II in endothelial NO production and endothelium-dependent relaxation
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DOI：
10.1152/ajpheart.00932.2001
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发表时间：
2003-06-01
影响因子：
4.8
通讯作者：
Dinh-Xuan, AT
中科院分区：
文献类型：
--
作者：
Schneider, JC;El Kebir, D;Dinh-Xuan, AT
一氧化氮 (NO) 由 L-精氨酸通过 Ca2+/钙调蛋白敏感的内皮 NO 合酶 (NOS) 亚型 (eNOS) 合成。本研究评估了 Ca2+/钙调蛋白依赖性蛋白激酶 II (CaMK II) 在内皮依赖性舒张和 NO 合成中的作用。研究了三种 CaMK II 抑制剂对血压正常大鼠内皮完整的主动脉环的影响。在用 Ca2+ 离子载体 A23187 和毒胡萝卜素刺激的培养的猪主动脉内皮细胞的培养基中，通过 NO 传感器和化学发光评估 NO 合成。通过将L-[H-3]精氨酸转化为L-[H-3]瓜氨酸来测量大鼠主动脉内皮NOS活性。三种 CaMK II 抑制剂（多肽 281-302、KN-93 和薰衣草素 C）可减弱内皮完整的大鼠主动脉环响应乙酰胆碱、A23187 和毒胡萝卜素的内皮依赖性松弛。 CaMK II 抑制剂均不影响 NO 供体诱导的松弛。在猪主动脉内皮细胞系中，KN-93 减少 NO 合成，并导致 A23187 和毒胡萝卜素的浓度-反应曲线右移。在大鼠主动脉内皮细胞中，KN-93 显着降低缓激肽诱导的 eNOS 活性。这些结果表明，由于 eNOS 的 Ca2+ 依赖性激活，CaMK II 参与了 NO 合成。
Nitric oxide (NO) is synthesized from L-arginine by the Ca2+/calmodulin-sensitive endothelial NO synthase (NOS) isoform (eNOS). The present study assesses the role of Ca2+/calmodulin-dependent protein kinase II (CaMK II) in endothelium-dependent relaxation and NO synthesis. The effects of three CaMK II inhibitors were investigated in endothelium-intact aortic rings of normotensive rats. NO synthesis was assessed by a NO sensor and chemiluminescence in culture medium of cultured porcine aortic endothelial cells stimulated with the Ca2+ ionophore A23187 and thapsigargin. Rat aortic endothelial NOS activity was measured by the conversion of L-[H-3] arginine to L-[H-3] citrulline. Three CaMK II inhibitors, polypeptide 281-302, KN-93, and lavendustin C, attenuated the endothelium-dependent relaxation of endothelium-intact rat aortic rings in response to acetylcholine, A23187, and thapsigargin. None of the CaMK II inhibitors affected the relaxation induced by NO donors. In a porcine aortic endothelial cell line, KN-93 decreased NO synthesis and caused a rightward shift of the concentration-response curves to A23187 and thapsigargin. In rat aortic endothelial cells, KN-93 significantly decreased bradykinin-induced eNOS activity. These results suggest that CaMK II was involved in NO synthesis as a result of Ca2+-dependent activation of eNOS.