Platelet and vascular thromboxane A2/prostaglandin H2 receptors. Evidence for different subclasses in the rat.
Thromboxane A2 (TXA2) and its precursor prostaglandin H2 (PGH2) induce platelet aggregation and vascular contraction through shared cell surface receptors commonly referred to as TXA2 or TXA2/PGH2 receptors. Whether different subclasses of TXA2/PGH2 receptors exist in platelets and vascular smooth muscle cells is controversial. In this study, TXA2 receptors on washed rat and human platelets and cultured rat aortic smooth muscle cells (RASMC) were characterized using radioligand competition binding assays with the 125I-labeled TXA2/PGH2 receptor agonist [1S-(1 alpha,2 beta(5Z),3 alpha(1E,3R*),4 alpha)] -7- [3-(3-hydroxy-4-(4'-iodophenoxy)-1-butenyl) -7- oxabicyclo-[2.2.1]- heptan-2-yl] -5- heptenoic acid (I-BOP) and various agonists and antagonists. Scatchard analyses of equilibrium binding data revealed Kd values of 205 +/- 68 pM (N = 6), 2.2 +/- 0.3 nM (N = 9) and 310 +/- 60 pM (N = 7) and Bmax values of 1.3 +/- 0.45 fmol/10(6) platelets, 2.8 +/- 0.2 fmol/10(6) platelets and 20.9 +/- 2.2 fmol/10(6) cells for rat and human platelets and RASMC, respectively. Concentration-dependent increases in intracellular free Ca2+ concentrations induced by I-BOP were observed in RASMC loaded with the calcium sensitive dye fura-2. The IC50 values for various TXA2/PGH2 analogues in competition binding assays with 125I-BOP were determined. Based on their IC50 values, the rank orders were I-BOP less than L657925 less than ONO11113 less than or equal to SQ29548 less than PTA-TPO less than PTA-NO less than or equal to L657926 less than or equal to I-PTA-OH less than PTA-OH[2] = meta-I-PTA-PO less than or equal to ONO11120[2] = ONO11120[1] less than PTA-OH[1] in rat platelets. I-BOP less than SQ29548 less than PTA-TPO = L657925 less than or equal to ONO11113 less than I-PTA-OH less than PTA-NO less than or equal to meta-I-PTA-PO less than or equal to PTA-OH[2] less than ONO11120[2] less than or equal to ONO11120[1] less than L657926 less than or equal to PTA-OH[1] in human platelets, and I-BOP less than L657925 less than ONO11113 less than or equal to SQ29548 less than ONO11120[2] less than or equal to L657926 less than or equal to PTA-OH[2] less than PTA-TPO less than ONO11120[1] less than I-PTA-OH less than meta-I-PTA-PO less than PTA-NO less than PTA-OH[1] in RASMC.(ABSTRACT TRUNCATED AT 400 WORDS)
Masuda
A
A
Department of Pharmacology, Medical University of South Carolina, Charleston 29425.
Mais
D E
DE
Oatis
J E
JE
Halushka
P V
PV
D000818:Animals; D001011:Aorta; D001667:Binding, Competitive; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism; D001643:Bridged Bicyclo Compounds / Q000494:pharmacology*; D019086:Bridged Bicyclo Compounds, Heterocyclic; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured / Q000187:drug effects; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology*; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000378:metabolism; D011869:Radioligand Assay; D051381:Rats; D011982:Receptors, Prostaglandin / Q000145:classification / Q000187:drug effects*; D017482:Receptors, Thromboxane; D044244:Receptors, Thromboxane A2, Prostaglandin H2; D013928:Thromboxane A2 / Q000031:analogs & derivatives / Q000494:pharmacology
D001643:Bridged Bicyclo Compounds; D019086:Bridged Bicyclo Compounds, Heterocyclic; D005231:Fatty Acids, Unsaturated; D011982:Receptors, Prostaglandin; D017482:Receptors, Thromboxane; D044244:Receptors, Thromboxane A2, Prostaglandin H2; C061750:7-(3-(3-hydroxy-4-(4'-iodophenoxy)-1-butenyl)-7-oxabicyclo(2.2.1)heptan-2-yl)-5-heptenoic acid; D013928:Thromboxane A2; D002118:Calcium