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Thromb Haemost 2012; 107(03): 521-529
DOI: 10.1160/TH11-07-0461
Platelets and Blood Cells
Schattauer GmbH

The thrombin inhibitors hirudin and Refludan® activate the soluble guanylyl cyclase and the cGMP pathway in washed human platelets

Anna Kobsar   *
1   Institut fuer Klinische Biochemie und Pathobiochemie-Zentrallabor, Universitaetsklinikum Wuerzburg, Wuerzburg, Germany
,
Juergen Koessler   *
1   Institut fuer Klinische Biochemie und Pathobiochemie-Zentrallabor, Universitaetsklinikum Wuerzburg, Wuerzburg, Germany
,
Linda Kehrer
1   Institut fuer Klinische Biochemie und Pathobiochemie-Zentrallabor, Universitaetsklinikum Wuerzburg, Wuerzburg, Germany
,
Stepan Gambaryan
1   Institut fuer Klinische Biochemie und Pathobiochemie-Zentrallabor, Universitaetsklinikum Wuerzburg, Wuerzburg, Germany
2   Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, 194223 St. Petersburg, Russia
,
Ulrich Walter
1   Institut fuer Klinische Biochemie und Pathobiochemie-Zentrallabor, Universitaetsklinikum Wuerzburg, Wuerzburg, Germany
› Author Affiliations Financial support: The study was supported by the Deutsche Forschungsgemeinschaft (SFB 688, TPA2, GA 1561/1–1).
Further Information

Publication History

Received: 07 July 2011

Accepted after major revision: 01 January 2011

Publication Date:
22 November 2017 (online)

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Summary

A number of direct thrombin inhibitors are successfully used clinically and experimentally as novel antithrombotics and specific anticoagulants. They are also used as anticoagulants in certain blood collection tubes for the analysis of platelet function. A series of platelet function tests have emerged to measure adequate responses to antiplatelet therapy. For comparative and practical reasons, it would be of advantage to use the same anticoagulant in blood collection tubes for different methods, e.g. thrombin inhibitors. However, there are little data on the effects of thrombin inhibitors on platelet signalling pathways that could influence results. We examined the applicability of thrombin inhibitor containing blood for platelet reactivity index (PRI) measurements of the VASP assay and investigated the effects of two thrombin inhibitors (hirudin and lepirudin) on cAMP- and cGMP-mediated signalling pathways in washed human platelets. We show that induction of VASP phosphorylation by PGE1 is markedly reduced in lepirudin containing blood samples. In consequence, PRI levels were highly variable compared to routinely used citrated blood. Surprisingly, in vitro incubation of platelets with thrombin inhibitors increases platelet cGMP levels and induces NOS independent sGC/PKG-mediated VASP phosphorylation. We conclude that thrombin inhibitors activate sGC/PKG-dependent pathways resulting in an increase of VASP phosphorylation which contributes to deviations in PRI measurements. These effects of thrombin inhibitors on sGC- and cGMP-mediated pathways including increased VASP phosphorylation may indicate the presence of an important additional platelet-based mechanism for the reduction of thrombus formation and thromboembolism by thrombin inhibitors.

Keywords

VASP - PRI - lepirudin - hirudin - sGC/ PKG/ cGMP pathway

* These authors contributed equally.


 

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