Collagen IV contributes to nitric oxide-induced angiogenesis of lung endothelial cells

Am J Physiol Cell Physiol. 2011 May;300(5):C979-88. doi: 10.1152/ajpcell.00368.2010. Epub 2011 Feb 9.


Nitric oxide (NO) mediates endothelial angiogenesis via inducing the expression of integrin α(v)β(3). During angiogenesis, endothelial cells adhere to and migrate into the extracellular matrix through integrins. Collagen IV binds to integrin α(v)β(3), leading to integrin activation, which affects a number of signaling processes in endothelial cells. In the present study, we evaluated the role of collagen IV in NO-induced angiogenesis. We found that NO donor 2,2'-(hydroxynitrosohydrazino)bis-ethanamine (NOC-18) causes increases in collagen IV mRNA and protein in lung endothelial cells and collagen IV release into the medium. Addition of collagen IV into the coating of endothelial culture increases endothelial monolayer wound repair, proliferation, and tube formation. Inhibition of collagen IV synthesis using gene silencing attenuates NOC-18-induced increases in monolayer wound repair, cell proliferation, and tube formation as well as in the phosphorylation of focal adhesion kinase (FAK). Integrin blocking antibody LM609 prevents NOC-18-induced increase in endothelial monolayer wound repair. Inhibition of protein kinase G (PKG) using the specific PKG inhibitor KT5823 or PKG small interfering RNA prevents NOC-18-induced increases in collagen IV protein and mRNA and endothelial angiogenesis. Together, these results indicate that NO promotes collagen IV synthesis via a PKG signaling pathway and that the increase in collagen IV synthesis contributes to NO-induced angiogenesis of lung endothelial cells through integrin-FAK signaling. Manipulation of collagen IV could be a novel approach for the prevention and treatment of diseases such as alveolar capillary dysplasia, severe pulmonary arterial hypertension, and tumor invasion.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carbazoles / pharmacology
  • Cells, Cultured
  • Collagen Type IV / antagonists & inhibitors
  • Collagen Type IV / metabolism
  • Collagen Type IV / pharmacology
  • Collagen Type IV / physiology*
  • Cyclic GMP-Dependent Protein Kinases / antagonists & inhibitors
  • Endothelial Cells / physiology*
  • Focal Adhesion Protein-Tyrosine Kinases / antagonists & inhibitors
  • Focal Adhesion Protein-Tyrosine Kinases / physiology
  • Integrins / antagonists & inhibitors
  • Integrins / physiology
  • Lung / blood supply*
  • Lung / drug effects
  • Lung / physiology
  • Neovascularization, Physiologic*
  • Nitric Oxide / agonists
  • Nitric Oxide / physiology*
  • Nitric Oxide Donors / pharmacology
  • Nitroso Compounds / pharmacology
  • Phosphorylation
  • Protein Kinase Inhibitors
  • RNA, Small Interfering / pharmacology
  • Swine


  • Carbazoles
  • Collagen Type IV
  • Integrins
  • NOC 18
  • Nitric Oxide Donors
  • Nitroso Compounds
  • Protein Kinase Inhibitors
  • RNA, Small Interfering
  • KT 5823
  • Nitric Oxide
  • Focal Adhesion Protein-Tyrosine Kinases
  • Cyclic GMP-Dependent Protein Kinases