title: PGE(2) reverses G(s)-mediated inhibition of platelet aggregation by interaction with EP3 receptors, but adds to non-G(s)-mediated inhibition of platelet aggregation by interaction with EP4 receptors.
abstract: Prostaglandin E(2) (PGE(2)) has intriguing effects on platelet function in the presence of agents that raise cyclic adenosine 3'5'-monophosphate (cAMP). PGE(2) reverses inhibition of platelet aggregation by agents that stimulate cAMP production via a G(s)-linked receptor, but adds to the inhibition of platelet function brought about by agents that raise cAMP through other mechanisms. Here, we used the EP receptor antagonists DG-041 (which acts at the EP3 receptor) and ONO-AE3-208 (which acts at the EP4 receptor) to investigate the role of these receptors in mediating these effects of PGE(2). Platelet aggregation was measured in platelet-rich plasma obtained from healthy volunteers in response to adenosine diphosphate (ADP) using single platelet counting. The effects of a range of concentrations of PGE(2) were determined in the presence of (1) the prostacyclin mimetic iloprost, which operates through G(s)-linked IP receptors, (2) the cAMP PDE inhibitor DN9693 and (3) the direct-acting adenylate cyclase stimulator forskolin. Vasodilator-stimulated phosphoprotein (VASP) phosphorylation was also determined as a measure of cAMP. PGE(2) reversed the inhibition of aggregation brought about by iloprost; this was prevented in the presence of the EP3 antagonist DG-041, indicating that this effect of PGE(2) is mediated via the EP3 receptor. In contrast, PGE(2) added to the inhibition of aggregation brought about by DN9693 or forskolin; this was reversed by the EP4 antagonist ONO-AE3-208, indicating that this effect of PGE(2) is mediated via the EP4 receptor. Effects on aggregation were accompanied by corresponding changes in VASP phosphorylation. The dominant role of EP3 receptors circumstances where cAMP is increased through a Gs-linked mechanism may be relevant to the situation in vivo where platelets are maintained in an inactive state through constant exposure to prostacyclin, and thus the main effect of PGE(2) may be prothrombotic. If so, the results described here further support the potential use of an EP3 receptor antagonist in the control of atherothrombosis.
authors.affiliation: School of Clinical Sciences, University of Nottingham, Nottingham, UK. jackie.glenn@nottingham.ac.uk
authors.forename: Jacqueline R
authors.initials: JR
authors.lastname: Glenn
authors.forename: Ann E
authors.initials: AE
authors.lastname: White
authors.forename: David
authors.initials: D
authors.lastname: Iyu
authors.forename: Stan
authors.initials: S
authors.lastname: Heptinstall
mesh_terms: D000178:Acrylamides / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects* / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000097:blood; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000097:blood; D015232:Dinoprostone / Q000494:pharmacology*; D006801:Humans; D008840:Microfilament Proteins / Q000097:blood; D009281:Naphthalenes / Q000494:pharmacology; D010654:Phenylbutyrates / Q000494:pharmacology; D010750:Phosphoproteins / Q000097:blood; D010766:Phosphorylation; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D011448:Prostaglandin Antagonists / Q000494:pharmacology; D011799:Quinazolines / Q000494:pharmacology; D043562:Receptors, G-Protein-Coupled / Q000097:blood*; D058308:Receptors, Prostaglandin E, EP3 Subtype / Q000037:antagonists & inhibitors / Q000097:blood*; D058309:Receptors, Prostaglandin E, EP4 Subtype / Q000037:antagonists & inhibitors / Q000097:blood*; D013450:Sulfones / Q000494:pharmacology; D000098662:Vasodilator-Stimulated Phosphoprotein
chemical_list: C531572:3-(1-((2,4-dichlorophenyl)methyl)-5-fluoro-3-methyl-1H-indol-7-yl)-N-((4,5-dichloro-2-thienyl)sulfonyl)-2-propenamide; C487890:4-(4-cyano-2-(2-(4-fluoronaphthalen-1-yl)propionylamino)phenyl)butyric acid; D000178:Acrylamides; D015815:Cell Adhesion Molecules; D008840:Microfilament Proteins; D009281:Naphthalenes; C545647:PTGER3 protein, human; D010654:Phenylbutyrates; D010750:Phosphoproteins; D010975:Platelet Aggregation Inhibitors; D011448:Prostaglandin Antagonists; D011799:Quinazolines; D043562:Receptors, G-Protein-Coupled; D058308:Receptors, Prostaglandin E, EP3 Subtype; D058309:Receptors, Prostaglandin E, EP4 Subtype; D013450:Sulfones; D000098662:Vasodilator-Stimulated Phosphoprotein; D005576:Colforsin; C049662:1,5-dihydro-7-(1-piperidinyl)-imidazo(2,1-b)quinazolin-2(3H)-one; D000242:Cyclic AMP; D015232:Dinoprostone