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Cardiovascular Research 2003 57(3):804-815; doi:10.1016/S0008-6363(02)00786-1
© 2003 by European Society of Cardiology
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Copyright © 2003, European Society of Cardiology

Crucial role of local peroxynitrite formation in neutrophil-induced endothelial cell activation

Hae-Young Sohna,*, Florian Krotzb, Stefan Zahlerb, Torsten Gloeb, Matthias Kellera, Karl Theisena, Thomas M Schielea, Volker Klaussa and Ulrich Pohlb

aDivision of Cardiology, Medizinische Klinik Innenstadt, Ludwig-Maximilians-University Munich, Ziemssenstrasse 1, 80336 Munich, Germany
bInstitute of Physiology, Ludwig-Maximilians-University Munich, Munich, Germany

* Corresponding author. Tel.: +49-89-5160-2266; fax: +49-89-5160-2410. sohn{at}lrz.uni-muenchen.de

Introduction and methods: The reaction of superoxide anions and NO not only results in a decreased availability of NO, but also leads to the formation of peroxynitrite, the role of which in the cardiovascular system is still discussed controversially. In cultured human endothelial cells, we studied whether there is a significant interaction between endothelial NO and neutrophil-derived superoxide anions in terms of endothelial peroxynitrite formation. We particularly studied whether a significantly higher redox-stress can be found in those endothelial cells directly adjacent to an activated neutrophil. Results: A considerable part of the 2,7-dihydrodichlorofluoresceine signal in endothelial cells was due to oxidation by peroxynitrite. Providing superoxide radicals by enzymatic source or by the neutrophil respiratory burst increased the fluorescence, which was attenuated by blockade of endothelial NO-synthase, suggesting that peroxynitrite was formed from neutrophil- or extracellular enzyme-derived superoxide and endothelial NO. Considerably higher fluorescence intensity was observed in endothelial cells in direct neighborhood to a neutrophil. This was particularly pronounced in the presence of a NO-donor and was accompanied by a strong activation of NF-{kappa}B and increased expression of E-selectin in these cells. Conclusion: Endothelial cells adjacent to neutrophils may have elevated levels of peroxynitrite that result in an increased expression of adhesion molecules. Such cells might represent a preferential site for adhesion and migration of additional neutrophils when simultaneously high concentrations of NO and neutrophil-derived superoxide are present.

KEYWORDS Endothelial function; Free radicals; Nitric oxide


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