Arachidonic acid-stimulated platelet tests: Identification of patients less sensitive to aspirin treatment

Temperilli, Flavia; Rina, Aldona; Massimi, Isabella; Montemari, Anna Lisa; Guarino, Maria; Zicari, Alessandra; Pulcinelli, Fabio M.

doi:10.3109/09537104.2014.1003291

Cited by 14 publications

(9 citation statements)

References 31 publications

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“…Platelet aggregation was evaluated in PRP in a four-channel aggregometer (AggRam, Helena Laboratories, Beaumont, Texas, United States) according toTemperilli et al 29 The results are reported as the percentage of aggregation observed after 4 minutes of stimulation (%PA) in response to ADP (2 μM) and primary maximal platelet aggregation in response to ADP (0.8 μM) (Helena Laboratories, Beaumont, Texas, United States). Results were considered statistically significant if a pvalue of less than 0.05 was reached.…”

Section: Platelet Aggregationmentioning

confidence: 99%

Aspirin-Dependent Effects on Purinergic P2Y1 Receptor Expression

et al. 2019

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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.

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Section: Platelet Aggregationmentioning

confidence: 99%

Aspirin-Dependent Effects on Purinergic P2Y1 Receptor Expression

et al. 2019

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“…In patients under chronic treatment, the compliance was assessed both by interview and as arachidonic acid-induced platelet aggregation (0.75 mmol/L) < 10%. 17 In our study, were excluded all those patients with cardiovascular diseases including: Ischemic Heart Disease (IHD), cerebrovascular disease or transient ischemic attack, established peripheral vascular disease, and class III to IV heart failure, in the previous 15 days.…”

Section: Patientsmentioning

confidence: 99%

MiR‐21 role in aspirin‐dependent PPARα and multidrug resistance protein 4 upregulation

Massimi

Alemanno

Guarino

et al. 2018

Research and Practice in Thrombosis and Haemostasis

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BackgroundA mechanism involved in high on‐aspirin treatment residual platelet reactivity is platelet multidrug resistance protein 4 (MRP4) overexpression. Aspirin enhances platelet MRP4 expression with a PPARα‐dependent mechanism and reduces miR‐21 expression that, in turn, downregulates PPARα expression.ObjectiveThe aim of our study was to verify the relationship between miR‐21 and MRP4‐PPARα levels induced by aspirin treatment.MethodsWe evaluated the changes in MRP4‐PPARα, mRNA, MRP4 protein, and miR‐21 expression induced by aspirin in: (i) in vitro–treated megakaryoblastic cell line (DAMI), (ii) primary megakaryocytes cultures and derived platelets, (iii) healthy volunteers’ platelets treated with aspirin, and (iv) aspirinated patients (aspirin‐treated patients) and in a control population (control).ResultsWe observed an aspirin‐induced reverse relationship between the expression of miR‐21 and PPARα‐MRP4. In DAMI cells the miR‐21 mimic transfection reduces PPARα and MRP4 expression, even if cells were treated with aspirin after transfection. MiR‐21 inhibitor transfection induces PPARα and MRP4 expression that are not enhanced by aspirin treatment. In human megakaryocytes, aspirin treatment lead to a miR‐21 downregulation and a MRP4 upregulation and this trend is confirmed in derived platelets. In aspirin‐treated volunteers, an inverse relationship between miR‐21 and MRP4 platelet expression was found after aspirin treatment. A similar negative relationship was found in aspirin‐treated patients vs the control population.ConclusionThe results reported in this study provide information that aspirin induces the modulation of platelet miR‐21 expression levels and this modulation can be responsible for MRP4 enhancement in circulating platelets.

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“…Platelet aggregation (PA) was performed as previously described [18]. PA was evaluated both in PRP and PFP using AggrRam aggregometer (Helena Laboratries, Beaumont, Texas, USA).…”

Section: Platelet Aggregationmentioning

confidence: 99%

MRP4 over-expression has a role on both reducing nitric oxide-dependent antiplatelet effect and enhancing ADP induced platelet activation

Guarino

Massimi

Alemanno

et al. 2020

J Thromb Thrombolysis

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The impact of inhibition of multidrug resistance protein 4 (MRP4) on nitric oxide (NO) resistance and on ADP-induced platelet aggregation is unknown. The aim of this investigation was to verify whether platelet NO resistance correlates with MRP4 expression and evaluate whether this can be reduced by in vitro MRP4 inhibition mediated by cilostazol. Moreover, we assessed if inhibition of MRP4-mediated transport reduces ADP-induced platelet reactivity. The inhibitory effect of sodium nitroprusside (SNP), a NO-donor that enhances cyclic guanosine monophosphate (cGMP) cytosolic concentration, was assessed in platelets obtained from aspirin treated patients and in a control population. The inhibitory effect of SNP was evaluated by ADP-induced aggregation in SNP-treated platelets. The impact of MRP4 on ADP-induced platelet aggregation was performed in high on aspirin residual platelet reactivity (HARPR) patients and compared to healthy volunteers (HV), and a control cohort (CTR). In aspirin-treated patients with high levels of MRP4, reduced SNP inhibition was found compared to those with low levels of MRP4. MRP4 inhibition by cilostazol significantly reduced ADP-induced platelet aggregation in HARPR population, and to a lesser extent in HV and CTR populations. In conclusion, cilostazol can mitigate the hyperreactive platelet phenotype of HARPR patients by reducing residual ADP-induced platelet aggregation and increasing NOdependent endothelial antiplatelet effects.

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Arachidonic acid-stimulated platelet tests: Identification of patients less sensitive to aspirin treatment

Cited by 14 publications

References 31 publications

Aspirin-Dependent Effects on Purinergic P2Y1 Receptor Expression

Aspirin-Dependent Effects on Purinergic P2Y1 Receptor Expression

MiR‐21 role in aspirin‐dependent PPARα and multidrug resistance protein 4 upregulation

MRP4 over-expression has a role on both reducing nitric oxide-dependent antiplatelet effect and enhancing ADP induced platelet activation

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