Nitric oxide is an upstream signal of vascular endothelial growth factor-induced extracellular signal-regulated kinase1/2 activation in postcapillary endothelium

J Biol Chem. 1998 Feb 13;273(7):4220-6. doi: 10.1074/jbc.273.7.4220.


We recently demonstrated that nitric oxide (NO) significantly contributes to the mitogenic effect of vascular endothelial growth factor (VEGF), suggesting a role for the NO pathway in the signaling cascade following kinase-derivative receptor activation in vascular endothelium. The aim of this study was to investigate the intracellular pathways linked to VEGF/NO-induced endothelial cell proliferation. We assessed the activity of the mitogen-activated protein kinase (MAPK) that is specifically activated by growth factors, extracellular-regulated kinase (ERK1/2), on cultured microvascular endothelium isolated from coronary postcapillary venules. ERK1/2 was immunoprecipitated, and its activity was assessed with an immunocomplex kinase assay. In endothelial cells exposed for 5 min to the NO donor drug sodium nitroprusside at a concentration of 100 microM, ERK1/2 activity significantly increased. VEGF produced a time- and concentration-dependent activation of ERK1/2. Maximal activity was obtained after 5 min of stimulation at a concentration of 10 ng/ml. The specific MAPK kinase inhibitor PD 98059 abolished ERK1/2 activation and endothelial cell proliferation in a concentration-dependent manner in response to VEGF and sodium nitroprusside. The NO synthase inhibitor Nomega-monomethyl-L-arginine, as well as the guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, blocked the activation of ERK1/2 induced by VEGF, suggesting that NO and cGMP contributed to the VEGF-dependent ERK1/2 activation. These results demonstrate for the first time that kinase-derivative receptor activation triggers the NO synthase/guanylate cyclase pathway to activate the MAPK cascade and substantiates the hypothesis that the activation of ERK1/2 is necessary for VEGF-induced endothelial cell proliferation.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Calcium / pharmacology
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism
  • Cell Division / physiology
  • Cells, Cultured
  • Endothelial Growth Factors / pharmacology*
  • Endothelium, Vascular / drug effects*
  • Endothelium, Vascular / growth & development
  • Enzyme Activation
  • Enzyme Inhibitors / pharmacology
  • Flavonoids
  • Guanylate Cyclase / antagonists & inhibitors
  • Lymphokines / pharmacology*
  • Mitogen-Activated Protein Kinase Kinases
  • Nitric Oxide / pharmacology*
  • Nitric Oxide Synthase / antagonists & inhibitors
  • Nitroprusside / pharmacology
  • Oxadiazoles / pharmacology
  • Protein Kinase Inhibitors
  • Quinoxalines / pharmacology
  • Receptor Protein-Tyrosine Kinases / physiology
  • Receptors, Growth Factor / physiology
  • Receptors, Vascular Endothelial Growth Factor
  • Signal Transduction / physiology
  • Vascular Endothelial Growth Factor A
  • Vascular Endothelial Growth Factors
  • omega-N-Methylarginine / pharmacology


  • 1H-(1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one
  • Endothelial Growth Factors
  • Enzyme Inhibitors
  • Flavonoids
  • Lymphokines
  • Oxadiazoles
  • Protein Kinase Inhibitors
  • Quinoxalines
  • Receptors, Growth Factor
  • Vascular Endothelial Growth Factor A
  • Vascular Endothelial Growth Factors
  • Nitroprusside
  • omega-N-Methylarginine
  • Nitric Oxide
  • Nitric Oxide Synthase
  • Receptor Protein-Tyrosine Kinases
  • Receptors, Vascular Endothelial Growth Factor
  • Calcium-Calmodulin-Dependent Protein Kinases
  • Mitogen-Activated Protein Kinase Kinases
  • Guanylate Cyclase
  • 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one
  • Calcium