title: Calcium-dependent synergistic interaction of platelet activating factor and epinephrine in human platelet aggregation.
abstract: OBJECTIVE
  To investigate the mechanism (s) involved in the synergistic interaction of platelet activating factor (PAF) and epinephrine.
  
  
  METHODS
  Blood was obtained from healthy human subjects reported to be free of medications for at least two weeks before sampling. Aggregation was monitored at 37 oC using Dual-channel Lumi-aggregometer. The resulting aggregation was recorded for 5 min by the measurement of light transmission as a function of time.
  
  
  RESULTS
  Platelet aggregation mediated by subthreshold concentrations of PAF (5-8 nmol/L) plus epinephrine (0.5-2 micromol/L) was inhibited by ?2-receptor blocker, yohimbine, and PAF receptor antagonist WEB 2086. This synergism was inhibited by calcium channel blockers, verapamil and diltiazem. In addition, platelet aggregation by co-addition of PAF and epinephrine was also inhibited by very low concentrations of phospholipase C (PLC) inhibitor (U73122; IC50=0.2 micromol/L), the MAP kinase inhibitor, PD 98059 (IC50=3 micromol/L), and cyclooxygenase (COX-1) inhibitors including indomethacin (IC50=0.25 micromol/L), flurbiprofen (IC50=0.7 micromol/L), and piroxicam (IC50=7 micromol/L). However, COX-2 inhibitors, nimesulide (IC50=26 micromol/L), NS-398 (IC50=7 micromol/L), and etodolac (IC50=15 micromol/L) were also effective in inhibiting the aggregation. The inhibitors of protein kinase C (chelerythrine) and tyrosine kinase (genistien), and phosphatidylinositol 3-kinase inhibitor (wortmannin) had no significant effect on platelet aggregation induced by PAF and epinephrine.
  
  
  CONCLUSIONS
  The synergistic effect of PAF and epinephrine on human platelet aggregation is receptor-mediated and involves the activation of PLC/Ca2+, COX and MAP kinase signalling pathways.
authors.affiliation: Department of Biological and Biomedical Sciences, The Aga Khan University, PO Box 3500, Stadium Road Karachi-74800, Pakistan. arshad.saeed@aku.edu
authors.forename: Sheikh Arshad
authors.initials: SA
authors.lastname: Saeed
authors.forename: Huma
authors.initials: H
authors.lastname: Rasheed
mesh_terms: D000316:Adrenergic alpha-Agonists / Q000494:pharmacology; D000317:Adrenergic alpha-Antagonists / Q000494:pharmacology; D001381:Azepines / Q000494:pharmacology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D004357:Drug Synergism; D004837:Epinephrine / Q000494:pharmacology*; D005480:Flurbiprofen / Q000494:pharmacology; D006801:Humans; D010972:Platelet Activating Factor / Q000494:pharmacology*; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D015398:Signal Transduction; D014230:Triazoles / Q000494:pharmacology; D014700:Verapamil / Q000494:pharmacology; D015016:Yohimbine / Q000494:pharmacology
chemical_list: D000316:Adrenergic alpha-Agonists; D000317:Adrenergic alpha-Antagonists; D001381:Azepines; D002121:Calcium Channel Blockers; D016861:Cyclooxygenase Inhibitors; D010972:Platelet Activating Factor; D010975:Platelet Aggregation Inhibitors; D014230:Triazoles; C052518:WEB 2086; D015016:Yohimbine; D005480:Flurbiprofen; D014700:Verapamil; D004837:Epinephrine