© 1998 by European Society of Cardiology
Copyright © 1998, European Society of Cardiology
Nitric oxide-dependent and independent effects on human platelets treated with peroxynitrite
aDepartments of Cardiology and Medicine, King's College Hospital, Denmark Hill, London SE5 9RS, UK
bDepartmento de Farmacolog
éa, Facultad de Medicina, Universidad Complutense de Madrid, E-28040 Madrid, Spain
cUnite INSERM U-91, Hopital Henri Mondor, Creteil, France
dDepartment of Pharmacology, University of Alberta, Edmonton, Canada
eDepartment of Pathology, Molecular and Cellular Division, Center for Free Radical Biology, University of Alabama at Birmingham, Volker Hall Room GO38, 1670 University Boulevard, Birmingham, AL 35294-0019, USA
* Corresponding author.
Objective: Peroxynitrite (ONOO–) is an oxidant formed from the rapid reaction of superoxide and nitric oxide (NO) at sites of inflammation. The literature reports conflicting data on the effects of ONOO– in biological systems, with both NO- and oxidant-dependent effects having been demonstrated. The aim of this study was to investigate these distinct mechanisms through examining molecular aspects of the effects of ONOO– on human platelets, a system in which we have previously shown that ONOO– has both pro- and anti-aggregatory effects. Methods: Platelet function was assessed by measuring platelet P-selectin expression flow cytometrically, intraplatelet Ca2+ concentrations, and by light aggregometry. A colorimetric method was used to measure extracellular platelet membrane thiols. The contribution of NO and cGMP to the pharmacological effects of ONOO– was investigated using an inhibitor of the soluble guanylate cyclase (sGC), 1H-[1,2,4]oxadiazolo[4,3-
]quinoxalin-1-one (ODQ), and the NO scavenger oxy-haemoglobin. Results: Peroxynitrite (50–400 µM) caused a concentration-dependent increase in the number of platelets expressing P-selectin, an increase in intraplatelet Ca2+ concentrations and a decrease in platelet membrane thiols. Peroxynitrite-induced P-selectin expression was augmented by ODQ. In contrast, when P-selectin expression was elicited by collagen, ONOO– acted as an inhibitor of this process, an effect that was further enhanced by the addition of 1% plasma. ODQ or oxy-haemoglobin abolished this inhibitory effect. Finally, low concentrations (50–100 µM) of ONOO– inhibited collagen-induced platelet aggregation, an effect that was reversed by oxy-haemoglobin. Conclusions: Peroxynitrite exerts dual effects on platelets, which are either activating or inhibitory due to the conversion of ONOO– to NO or NO donors. Peroxynitrite-induced platelet activation seems to be due to thiol oxidation and an increase in intracellular Ca2+. It is important to note that inhibitory, NO-dependent effects occur at lower concentrations than the activating effects. These data are then consistent with the conflicting literature, showing both damaging and cytoprotective effects of ONOO– in biological systems. We hypothesize that the conversion of ONOO– to NO is the critical factor determining the outcome of ONOO– exposure in vivo.
KEYWORDS Platelets; Nitric oxide; Peroxynitrite; P-selectin; cGMP
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  S. R. Thom, V. M. Bhopale, S.-T. Han, J. M. Clark, and K. R. Hardy Intravascular Neutrophil Activation Due to Carbon Monoxide Poisoning Am. J. Respir. Crit. Care Med., December 1, 2006; 174(11): 1239 - 1248. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Marcondes, M. H. M. Cardoso, R. P. Morganti, S. M. Thomazzi, S. Lilla, F. Murad, G. De Nucci, and E. Antunes Cyclic GMP-independent mechanisms contribute to the inhibition of platelet adhesion by nitric oxide donor: A role for {alpha}-actinin nitration PNAS, February 28, 2006; 103(9): 3434 - 3439. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Chung, S. M. Wildhirt, S. Wang, A. Koshal, and M. W. Radomski Combined administration of nitric oxide gas and iloprost during cardiopulmonary bypass reduces platelet dysfunction: A pilot clinical study J. Thorac. Cardiovasc. Surg., April 1, 2005; 129(4): 782 - 790. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Krotz, H.-Y. Sohn, and U. Pohl Reactive Oxygen Species: Players in the Platelet Game Arterioscler Thromb Vasc Biol, November 1, 2004; 24(11): 1988 - 1996. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Muzaffar, N. Shukla, G. Angelini, and J. Y. Jeremy Nitroaspirins and Morpholinosydnonimine but Not Aspirin Inhibit the Formation of Superoxide and the Expression of gp91phox Induced by Endotoxin and Cytokines in Pig Pulmonary Artery Vascular Smooth Muscle Cells and Endothelial Cells Circulation, August 31, 2004; 110(9): 1140 - 1147. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. K. Gill, S. Saksena, I. A. Syed, S. Tyagi, W. A. Alrefai, J. Malakooti, K. Ramaswamy, and P. K. Dudeja Regulation of NHE3 by nitric oxide in Caco-2 cells Am J Physiol Gastrointest Liver Physiol, September 1, 2002; 283(3): G747 - G756. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Ferretti, R. A. Rabini, T. Bacchetti, A. Vignini, E. Salvolini, F. Ravaglia, G. Curatola, and L. Mazzanti Glycated Low Density Lipoproteins Modify Platelet Properties: A Compositional and Functional Study J. Clin. Endocrinol. Metab., May 1, 2002; 87(5): 2180 - 2184. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Coles, A. Bloodsworth, J. P. Eiserich, M. J. Coffey, R. M. McLoughlin, J. C. Giddings, M. J. Lewis, R. J. Haslam, B. A. Freeman, and V. B. O'Donnell Nitrolinoleate Inhibits Platelet Activation by Attenuating Calcium Mobilization and Inducing Phosphorylation of Vasodilator-stimulated Phosphoprotein through Elevation of cAMP J. Biol. Chem., February 15, 2002; 277(8): 5832 - 5840. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. R. Thom, D. Fisher, and Y. Manevich Roles for platelet-activating factor and {middle dot}NO-derived oxidants causing neutrophil adherence after CO poisoning Am J Physiol Heart Circ Physiol, August 1, 2001; 281(2): H923 - H930. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. KOBAYASHI, E. C. GABAZZA, S. SHIMIZU, H. YASUI, H. YUDA, O. HATAJI, K. MARUYAMA, T. YAMAUCHI, K. SUZUKI, Y. ADACHI, et al. Long-term Inhalation of High-dose Nitric Oxide Increases Intraalveolar Activation of Coagulation System in Mice Am. J. Respir. Crit. Care Med., June 1, 2001; 163(7): 1676 - 1682. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. P. Patel, A.-L. Levonen, J. H. Crawford, and V. M. Darley-Usmar Mechanisms of the pro- and anti-oxidant actions of nitric oxide in atherosclerosis Cardiovasc Res, August 18, 2000; 47(3): 465 - 474. [Abstract] [Full Text] [PDF]
|
|