Anti-oxidant ebselen delays microvascular thrombus formation in the rat cremaster muscle by inhibiting platelet P-selectin expression

Nicole Lindenblatt; Wolfgang Schareck; Lorenz Belusa; Ruth Maria Nickels; Michael Dieter Menger; Brigitte Vollmar

doi:10.1160/TH02-09-0093

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2003; 90(05): 882-892
DOI: 10.1160/TH02-09-0093

Platelets and Blood Cells

Schattauer GmbH

Anti-oxidant ebselen delays microvascular thrombus formation in the rat cremaster muscle by inhibiting platelet P-selectin expression

Nicole Lindenblatt

¹Department of Experimental Surgery, University of Rostock, Germany

²Department of General Surgery, University of Rostock, Germany

,

Wolfgang Schareck

²Department of General Surgery, University of Rostock, Germany

,

Lorenz Belusa

²Department of General Surgery, University of Rostock, Germany

,

Ruth Maria Nickels

³Department of Clinical-Experimental Surgery, University of Saarland, Homburg/Saar, Germany

,

Michael Dieter Menger

³Department of Clinical-Experimental Surgery, University of Saarland, Homburg/Saar, Germany

,

Brigitte Vollmar

¹Department of Experimental Surgery, University of Rostock, Germany

› Author Affiliations

Abstract

Summary

Ebselen, a seleno-organic compound showing glutathione peroxidase-like activity, has potent anti-inflammatory and anti-oxidant effects. Since selenium deficiency is thought to be associated with an increased incidence of vascular thrombosis, we studied the effect of ebselen on blood cell aggregate formation and vessel occlusion in vivo. In individual microvessels of rat cremaster muscle preparations, photochemically induced thrombus formation was analyzed in detail using intravital fluorescence microscopy. In ebselen-pretreated animals (30mg/kg ip), venular thrombus formation was significantly delayed (50% vessel occlusion: 535±34s; initial stasis: 872±82s; complete occlusion: 908±87s) as compared to vehicle-treated controls (416±42; 612±49; 647±51). Moreover, ebselen significantly prolonged the kinetics of arteriolar thrombus formation and even completely prevented blood cell aggregate and thrombus formation in 88.9% of all arterioles studied (p<0.05 vs controls:37.5%). Anti-thrombotic properties of ebselen could also be observed in a model of ferric chloride-induced microvascular thrombosis, with a low dose (5mg/kg ip) being as effective as a high dose pretreatment (30mg/kg ip). As assessed by flow cytometry of platelet P-selectin immunfluorescence, whole blood isolated from ebselen-treated animals revealed a significantly lower fraction of P-selectin expressing platelets when compared with that of DMSO-treated controls. In addition, oxidant stress-induced upregulation of P-selectin on isolated platelets was found dose-dependently inhibited by increasing concentrations of ebselen (10-100µM). Moreover, ebselen dose-dependently inhibited H₂O₂-induced platelet-leukocyte aggregate formation in whole blood in vitro, suggesting that the anti-thrombotic effect of ebselen is achieved by attenuation of P-selectin dependent platelet-leukocyte aggregation. Thus, ebselen represents preventive and therapeutic value for disorders with increased risk for oxidant stress-associated thrombotic events.

Keywords

Thrombosis - platelet - leukocyte - fluorescence microscopy - flow cytometry - oxidant stress

Full Text

References

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