Involvement of thromboxane A2 and tyrosine kinase in the synergistic interaction of platelet activating factor and calcium ionophore A23187 in human platelet aggregation.
The present study was carried out to examine the mechanisms of the synergistic interaction of PAF and A23187 mediated platelet aggregation. We found that platelet aggregation mediated by subthreshold concentrations of PAF (5 nM) and A23187 (1 mM) was inhibited by PAF receptor blocker (WEB 2086, IC50 = 0.65 mM) and calcium channel blockers, diltiazem (IC50 = 13 mM) and verapamil (IC50 = 18 mM). Pretreatment of platelets with PAF and A23187 induced rise in intracellular calcium and this effect was also blocked by verapamil. While examining the role of the down stream signaling pathways, we found that platelet aggregation induced by the co-addition of PAF and A23187 was also inhibited by low concentrations of phospholipase C (PLC) inhibitor (U73122; IC50 = 10 mM), a cyclooxygenase inhibitor (indomethacin; IC50 = 0.2 mM) and inhibitor of TLCK, herbimycin A with IC50 value of 5 mM. The effect was also inhibited by a specific TXA2 receptor antagonist, SQ 29548 with very low IC50 value of 0.05 mM. However, the inhibitors of MAP kinase, PD98059 and protein kinase C, chelerythrine had no effect on PAF and A23187-induced platelet aggregation. These data suggest that the synergism between PAF and A23187 in platelet aggregation involves activation of thromboxane and tyrosine kinase pathways.
Department of Biological and Biomedical Sciences, The Aga Khan University, Karachi-74800, Pakistan.
Huma
H
Rasheed
Sheikh Arshad
SA
Saeed
D016227:Benzoquinones; D001792:Blood Platelets / Q000187:drug effects*; D000001:Calcimycin / Q000494:pharmacology*; D006801:Humans; D007213:Indomethacin / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D047029:Lactams, Macrocyclic; D010972:Platelet Activating Factor / Q000378:metabolism / Q000494:pharmacology*; D010974:Platelet Aggregation / Q000502:physiology*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D011809:Quinones / Q000494:pharmacology; D017828:Rifabutin / Q000031:analogs & derivatives; D013928:Thromboxane A2 / Q000502:physiology*; D014700:Verapamil / Q000494:pharmacology
D016227:Benzoquinones; D007476:Ionophores; D047029:Lactams, Macrocyclic; D010972:Platelet Activating Factor; D011809:Quinones; D017828:Rifabutin; D000001:Calcimycin; D013928:Thromboxane A2; C020754:herbimycin; D014700:Verapamil; D011505:Protein-Tyrosine Kinases; D007213:Indomethacin