Synergism interaction between arachidonic acid by 5-hydroxytryptamine in human platelet aggregation is mediated through multiple signalling pathways
Document Type
Article
Department
Biological and Biomedical Sciences
Abstract
AIM: To examine the signalling mechanisms involved in the synergistic interaction of 5-hydroxytryptamine (5-HT) and arachidonic acid (AA) in human platelet aggregation.Methods: Blood was obtained from healthy human subjects, mixed with 3.8 % sodium citrate (9:1), and centrifuged to prepare platelet rich plasma (PRP). Aggregation was monitored using a Dual-channel Lumi-aggregometer. The agonist-induced influx of Ca2+ was measured using Fura-2 AM. TXA2 formation was studied using radiochemical method.Results: Subthreshold concentration of 5-HT (2 micromol/L) potentiated the effect of low dose of AA (0.2 mmol/L) in human platelets. This synergistic effect was blocked by 5-HT2 receptor antagonist (methysergide IC(50)=5.2 nmol/L; cyproheptadine IC(50)=0.6 nmol/L), and thromboxane A2 receptor antagonist (SQ 29 548; IC(50)=30 nmol/L), showing that the effect is receptor-mediated. To examine the down-stream signalling pathways, we found that such an interaction was inhibited by calcium channel blockers (diltiazem; IC(50)=3 micromol/L and verapamil; IC(50)=5 micromol/L), phospholipase C (PLC) inhibitor (U73122; IC50=4 micromol/L), cyclooxygenase inhibitor, (indomethacin; IC(50)=0.2 micromol/L) and mitogen-activated protein (MAP) kinase inhibitor (PD98059; IC(50)=3 micromol/L). The effect was also inhibited by a specific tyrosine light chain kinase (TLCK) inhibitor, herbimycin A with IC(50) value of 5 micromol/L. Pretreatment of platelet with 5-HT and AA induced rise in intracellular calcium and this effect was blocked by verapamil.CONCLUSION: The synergism between 5-HT and AA in platelet aggregation involves activation of PLC/Ca2+, COX, and MAP kinase pathways.
Publication (Name of Journal)
Acta Pharmacologica Sinica
Recommended Citation
Saeed, S. A.,
Rasheed, H.,
Gilani, A.
(2003). Synergism interaction between arachidonic acid by 5-hydroxytryptamine in human platelet aggregation is mediated through multiple signalling pathways. Acta Pharmacologica Sinica, 24(10), 958-964.
Available at:
https://ecommons.aku.edu/pakistan_fhs_mc_bbs/660