Exogenous nitric oxide increases neutrophil adhesion to cultured human endothelial monolayers through a protein kinase G dependent mechanism

Inflammation. 1999 Feb;23(1):37-50. doi: 10.1023/a:1020287516911.


Endothelial-neutrophil adhesion is a critical step in acute inflammatory diseases, which is mediated in part by P-selectin and platelet-activating factor (PAF). Nitric oxide (NO) is well known as an endogenous second messenger derived from endothelial cells, and regulates many important physiological events, however, the direct effects of NO on endothelial-neutrophil adhesion is less well understood. The objective of this study was to examine whether, and how relatively high levels of exogenous NO increases neutrophil adhesion with respect to P-selectin and PAF. Endothelial monolayers were exposed to chemical agents for 30 min, and the adhesion of 51Cr-labeled neutrophils measured in a static adhesion assay. Spermine-NONOate (SNO), an NO donor, significantly increased neutrophil adhesion and expression of P-selectin at a concentration of 1 mM. SNO (1 mM)-mediated neutrophil adhesion was significantly inhibited by a protein kinase G inhibitor, KT5823 (0.5 microM), but not by a classical protein kinase C inhibitor, Gö6976 (10 nM), a tyrosine kinase inhibitor, genistein (1 microM), or a protein kinase A inhibitor, H-89 (0.1 microM). P-selectin surface expression induced by 1 mM SNO was also significantly inhibited by 0.5 microM KT5823. Conversely, a cytoplasm calcium chelator, TMB-8 (0.1 mM), significantly exacerbated both the neutrophil adhesion and P-selectin expression induced by SNO. WEB 2086 (10 microM), a PAF receptor antagonist, blocked neutrophil adhesion, but did not block P-selectin expression induced by SNO. These data suggest that NO increases endothelial-neutrophil adhesion through protein kinase G-mediated P-selectin mobilization to the cell surface and endothelial PAF synthesis.

Publication types

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

MeSH terms

  • Azepines / pharmacology
  • Cell Adhesion / drug effects
  • Cell Adhesion / physiology
  • Cells, Cultured
  • Cyclic GMP-Dependent Protein Kinases
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / metabolism
  • Endothelium, Vascular / physiology*
  • Humans
  • Intracellular Membranes / physiology
  • Neutrophils / drug effects*
  • Neutrophils / physiology*
  • Nitric Oxide / pharmacology*
  • Nitric Oxide Donors / pharmacology
  • Nitrogen Oxides
  • P-Selectin / metabolism
  • Platelet Aggregation Inhibitors / pharmacology
  • Protein Kinases / physiology*
  • Second Messenger Systems / physiology
  • Spermine / analogs & derivatives
  • Spermine / pharmacology
  • Triazoles / pharmacology


  • Azepines
  • Nitric Oxide Donors
  • Nitrogen Oxides
  • P-Selectin
  • Platelet Aggregation Inhibitors
  • Triazoles
  • WEB 2086
  • spermine nitric oxide complex
  • Spermine
  • Nitric Oxide
  • Protein Kinases
  • Cyclic GMP-Dependent Protein Kinases