Intracellular mechanisms of nitric oxide plus hydrogen peroxide-mediated neutrophil adherence to cultured human endothelial cells.
OBJECTIVE
We investigated signal-transduction in nitric oxide/hydrogen peroxide (NO/H2O2) mediated neutrophil-endothelial adhesion and P-selectin mobilization.


METHODS
Human endothelial monolayers (HUVEC) were exposed to 0.1 mM H2O2 plus an NO donor, 0.5 mM spermine-NONOate, and second message inhibitors and neutrophil adhesion and P-selectin expression measured.


RESULTS
Neutrophil adherence induced by NO/H2O2 was blocked by a PKG inhibitor, (KT5823, 0.5 microM), a PKC inhibitor, (Go6976, 10 nM), a calcium chelator, TMB-8 (0.1 mM) and a K+ channel blocker, glibenclamide, (10 microM), but not by a PKA inhibitor, (H-89, 0.1 microM) or a tyrosine kinase inhibitor, (genistein, I microM). P-selectin expression induced by NO/H2O2 was blocked by KT5823 and Gö6976, but not by TMB-8 or glibenclamide.


CONCLUSIONS
These data demonstrate that NO/ H2O2 promotes neutrophil-endothelial adhesion through PKG, PKC, calcium, and K+ channels, but not PKA or tyrosine kinase. Conversely, P-selectin mobilization requires only PKG and PKC.
Okayama
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N
Department of Molecular and Cellular Physiology, Louisiana State University Medical Center, Shreveport 71130-3932, USA.
Coe
L
L
Itoh
M
M
Alexander
J S
JS
D002118:Calcium / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects*; D002478:Cells, Cultured; D017869:Cyclic GMP-Dependent Protein Kinases; D004730:Endothelium, Vascular / Q000166:cytology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005905:Glyburide / Q000494:pharmacology; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D009504:Neutrophils / Q000502:physiology*; D009569:Nitric Oxide / Q000494:pharmacology*; D019007:P-Selectin / Q000378:metabolism; D026902:Potassium Channel Blockers; D015221:Potassium Channels / Q000502:physiology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000378:metabolism; D015290:Second Messenger Systems; D015398:Signal Transduction; D014471:Umbilical Veins
D004791:Enzyme Inhibitors; D019007:P-Selectin; D026902:Potassium Channel Blockers; D015221:Potassium Channels; D047428:Protein Kinase Inhibitors; D009569:Nitric Oxide; D006861:Hydrogen Peroxide; D011494:Protein Kinases; D017869:Cyclic GMP-Dependent Protein Kinases; D011493:Protein Kinase C; D005905:Glyburide; D002118:Calcium