Thromb Haemost 2019; 119(05): 726-734
DOI: 10.1055/s-0039-1678707
Cellular Haemostasis and Platelets
Georg Thieme Verlag KG Stuttgart · New York

Aspirin-Dependent Effects on Purinergic P2Y1 Receptor Expression

Isabella Massimi
1  Department of Experimental Medicine, ‘Sapienza’ University of Rome, Rome, Italy
,
Laura Alemanno
1  Department of Experimental Medicine, ‘Sapienza’ University of Rome, Rome, Italy
,
Maria Luisa Guarino
1  Department of Experimental Medicine, ‘Sapienza’ University of Rome, Rome, Italy
,
Raffaella Guerriero
2  Department of Cardiovascular, Dysmetabolic and Aging-associated Diseases, Istituto Superiore di Sanità, Rome, Italy
,
Massimo Mancone
3  Department of Cardiovascular, Respiratory, Nephrology, Anesthesiology and Geriatric Sciences, ‘Sapienza’ University of Rome, Rome, Italy
,
Andrea Ceccacci
3  Department of Cardiovascular, Respiratory, Nephrology, Anesthesiology and Geriatric Sciences, ‘Sapienza’ University of Rome, Rome, Italy
,
Luigi Frati
4  I.R.C.C.S Neuromed, Località Camerelle, Pozzilli (CB), Italy
,
Dominick J. Angiolillo
5  Division of Cardiology, University of Florida College of Medicine-Jacksonville, Florida, United States
,
Fabio M. Pulcinelli
1  Department of Experimental Medicine, ‘Sapienza’ University of Rome, Rome, Italy
› Author Affiliations
Funding This study was supported by a grant from ‘Sapienza’ - University of Rome 2015 to F.M.P.
Further Information

Publication History

23 July 2018

02 January 2019

Publication Date:
13 February 2019 (eFirst)

Abstract

Chronic treatment with aspirin in healthy volunteers (HVs) is associated with recovery of adenosine diphosphate (ADP)-induced platelet activation. The purinergic P2Y1 receptor exerts its effects via a Gq-protein, which is the same biochemical pathway activated by thromboxane-A2 receptor. We hypothesized that recovery of ADP-induced platelet activation could be attributed to increased P2Y1 expression induced by chronic aspirin exposure. We performed a multi-phase investigation which embraced both in vitro and in vivo experiments conducted in (1) human megakaryoblastic DAMI cells, (2) human megakaryocytic progenitor cell cultures, (3) platelets obtained from HVs treated with aspirin and (4) platelets obtained from aspirin-treated patients. DAMI cells treated with aspirin or WY14643 (PPARα agonist) had a significant up-regulation of P2Y1 mRNA, which was shown to be a PPARα-dependent process. In human megakaryocytic progenitors, in the presence of aspirin or WY14643, P2Y1 mRNA expression was higher than in mock culture. P2Y1 expression increased in platelets obtained from HVs treated with aspirin for 8 weeks. Platelets obtained from patients who were on aspirin for more than 2 months had increased P2Y1 expression and ADP-induced aggregation compared with patients on aspirin treatment for less than a month. Overall, our results suggest that aspirin induces genomic changes in megakaryocytes leading to P2Y1 up-regulation and that PPARα is the nuclear receptor involved in this regulation. Since P2Y1 is coupled to the same Gq-protein of thromboxane-A2 receptor, platelet adaptation in response to pharmacological inhibition seems not to be receptor specific, but may involve other receptors with the same biochemical pathway.

Authors' Contributions

I.M. designed the study, performed experiments, analysed results and wrote the manuscript. L.A., M.L.G. and R.G. contributed to the performed experiment and analysed results. L.F. and D.J.A. contributed in the data interpretation and revision of the intellectual content of the manuscript. M.M. and A.C. contributed to the data collection and patient enrolment. F.M.P. provided the concept of the study, designed the study, analysed results, and wrote the manuscript.