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Am J Physiol Heart Circ Physiol 284: H2311-H2319, 2003. First published January 30, 2003; doi:10.1152/ajpheart.00932.2001
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Vol. 284, Issue 6, H2311-H2319, June 2003

Involvement of Ca2+/calmodulin-dependent protein kinase II in endothelial NO production and endothelium-dependent relaxation

Jean-Christophe Schneider1, Driss El Kebir1, Christiane Chéreau2, Sophie Lanone3, Xiao-Lin Huang1, Anthony S. De Buys Roessingh1, Jean-Christophe Mercier4, Josette Dall'Ava-Santucci1, and A. Tuan Dinh-Xuan1

1 Service de Physiologie-Explorations Fonctionnelles, Centre Hospitalier Universitaire Cochin, Assistance Publique, Hôpitaux de Paris, Université Paris 5, 75014 Paris; 2 Laboratoire d'Immunologie Biologique, Faculté de Médecine Cochin, Université Paris 5, 75014 Paris; 3 Unité 408, Institut National de la Santé et de la Recherche Médicale, 75018 Paris; and 4 Service de Réanimation Pédiatrique, Hôpital Robert Debré, Assistance Publique, Hôpitaux de Paris, 75019 Paris, France

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-[3H]arginine to L-[3H]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.

endothelial function; nitric oxide; protein phosphorylation; signal transduction; thapsigargin


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