VEGF upregulates ecNOS message, protein, and NO production in human endothelial cells.

Vascular endothelial growth factor (VEGF) is an endothelium-specific secreted protein that potently stimulates vasodilation, microvascular hyperpermeability, and angiogenesis. Nitric oxide (NO) is also reported to modulate vascular tone, permeability, and capillary growth. Therefore, we hypothesized that VEGF might regulate endothelial production of NO. The production of nitrogen oxides by human umbilical vein endothelial cells (HUVECs) was measured after 1, 12, 24, and 48 h of incubation with VEGF. VEGF treatment resulted in both an acute (1 h) and chronic (> 24 h) stimulation of NO production. Furthermore, Western and Northern blotting revealed a VEGF-elicited, dose-dependent increase in the cellular content of endothelial cell nitric oxide synthase (ecNOS) message and protein that may account for the chronic upregulation of NO production elicited by VEGF. Finally, endothelial cells pretreated with VEGF for 24 h and subsequently exposed to A-23187 for 1 h produced NO at approximately twice the rate of cells that were not pretreated with VEGF. We conclude that VEGF upregulates ecNOS enzyme and elicits a biphasic stimulation of endothelial NO production.

[1]  H. Granger,et al.  Nitric oxide promotes proliferation and plasminogen activator production by coronary venular endothelium through endogenous bFGF. , 1997, Circulation research.

[2]  D. Bates,et al.  Vascular endothelial growth factor increases hydraulic conductivity of isolated perfused microvessels. , 1996, The American journal of physiology.

[3]  H. Granger,et al.  VEGF induces NO-dependent hyperpermeability in coronary venules. , 1996, The American journal of physiology.

[4]  B. Keyt,et al.  Effects of vascular endothelial growth factor on hemodynamics and cardiac performance. , 1996, Journal of cardiovascular pharmacology.

[5]  K. Thomas.  Vascular Endothelial Growth Factor, a Potent and Selective Angiogenic Agent (*) , 1996, The Journal of Biological Chemistry.

[6]  J. Liao,et al.  Regulation of bovine endothelial constitutive nitric oxide synthase by oxygen. , 1995, The Journal of clinical investigation.

[7]  R M Nerem,et al.  Regulation of endothelial cell nitric oxide synthase mRNA expression by shear stress. , 1995, The American journal of physiology.

[8]  D. Harrison,et al.  Molecular regulation of the bovine endothelial cell nitric oxide synthase by transforming growth factor-beta 1. , 1995, Arteriosclerosis, thrombosis, and vascular biology.

[9]  K. Rakušan.  Coronary Angiogenesis , 1995, Annals of the New York Academy of Sciences.

[10]  D. Harrison,et al.  Regulation of endothelial nitric oxide synthase mRNA, protein, and activity during cell growth. , 1994, The American journal of physiology.

[11]  S. Liu,et al.  Vascular endothelial growth factor induces EDRF-dependent relaxation in coronary arteries. , 1993, The American journal of physiology.

[12]  H. Granger,et al.  Nitric oxide promotes DNA synthesis and cyclic GMP formation in endothelial cells from postcapillary venules. , 1993, Biochemical and biophysical research communications.

[13]  H. Granger,et al.  Histamine increases venular permeability via a phospholipase C-NO synthase-guanylate cyclase cascade. , 1993, The American journal of physiology.

[14]  E. Keshet,et al.  Vascular endothelial growth factor induced by hypoxia may mediate hypoxia-initiated angiogenesis , 1992, Nature.

[15]  D. Harrison,et al.  Vasorelaxant properties of the endothelium-derived relaxing factor more closely resemble S-nitrosocysteine than nitric oxide , 1990, Nature.

[16]  D. Harrison,et al.  Release of NO and EDRF from cultured bovine aortic endothelial cells. , 1989, The American journal of physiology.

[17]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[18]  H. Granger,et al.  Nitric oxide mediates mitogenic effect of VEGF on coronary venular endothelium. , 1996, The American journal of physiology.

[19]  J. Folkman.  Angiogenesis in cancer, vascular, rheumatoid and other disease , 1995, Nature Medicine.

[20]  D. Harrison,et al.  The endothelial cell nitric oxide synthase: is it really constitutively expressed? , 1995, Agents and actions. Supplements.

[21]  T. Quertermous,et al.  Cloning and expression of a cDNA encoding human endothelium-derived relaxing factor/nitric oxide synthase. , 1992, The Journal of biological chemistry.

[22]  H. Dvorak,et al.  Tumor-secreted vascular permeability factor increases cytosolic Ca2+ and von Willebrand factor release in human endothelial cells. , 1991, The American journal of pathology.