The Phosphatase Inhibitor Calyculin-A Impairs Clot Retraction, Platelet Activation, and Thrombin Generation.

Hudák, Renáta; Vincze, János; Csernoch, László; Beke Debreceni, Ildikó; Oláh, Tamás; Erdődi, Ferenc; Clemetson, Kenneth John; Kappelmayer, János (2017). The Phosphatase Inhibitor Calyculin-A Impairs Clot Retraction, Platelet Activation, and Thrombin Generation. BioMed research international, 2017(9795271), p. 9795271. Hindawi Publishing Corporation 10.1155/2017/9795271

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The aim of this study was to investigate the effect of the serine/threonine protein phosphatase inhibitor, calyculin-A (CLA), on clot formation and on the procoagulant activity of human platelets. Platelet-rich plasma (PRP) samples were preincubated with buffer or CLA and subsequently platelets were activated by the protease-activated receptor 1 (PAR-1) activator, thrombin receptor activating peptide (TRAP). Clot retraction was detected by observing clot morphology up to 1 hour, phosphatidylserine- (PS-) expression was studied by flow cytometry, and thrombin generation was measured by a fluorimetric assay. For the intracellular Ca(2+) assay, platelets were loaded with calcium-indicator dyes and the measurements were carried out using a ratiometric method with real-time confocal microscopy. CLA preincubation inhibited clot retraction, PS-expression, and thrombin formation. TRAP activation elicited Ca(2+) response and PS-expression in a subset of platelets. The activated PRP displayed significantly faster and enhanced thrombin generation compared to nonactivated samples. CLA pretreatment abrogated PS-exposure and clot retraction also in TRAP-activated samples. As a consequence of the inhibitory effect on calcium elevation and PS-expression, CLA significantly downregulated thrombin generation in PRP. Our results show that CLA pretreatment may be a useful tool to investigate platelet activation mechanisms that contribute to clot formation and thrombin generation.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Department of Haematology, Oncology, Infectious Diseases, Laboratory Medicine and Hospital Pharmacy (DOLS) > Clinic of Haematology and Central Haematological Laboratory
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Unit Childrens Hospital > Forschungsgruppe Hämatologie (Erwachsene)

UniBE Contributor:

Clemetson, Kenneth John

Subjects:

600 Technology > 610 Medicine & health

ISSN:

2314-6133

Publisher:

Hindawi Publishing Corporation

Language:

English

Submitter:

Katrin Kölliker-Schütz

Date Deposited:

08 Dec 2017 15:27

Last Modified:

23 Oct 2019 21:36

Publisher DOI:

10.1155/2017/9795271

PubMed ID:

28680886

BORIS DOI:

10.7892/boris.106217

URI:

https://boris.unibe.ch/id/eprint/106217

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