The central role of thromboxane and platelet activating factor receptors in ex vivo regulation of endotoxin-induced monocyte tissue factor activity in human whole blood.
Expression of tissue factor (TF) by activated monocytes may initiate thrombotic episodes associated with diseases, such as thrombosis and atherosclerosis. In this study, steps in the regulatory pathways of lipopolysaccharide (LPS)-induced monocyte TF activity and released TNF-alpha in human whole blood were probed for using an array of inhibitors, comprising specific inhibitors of cytosolic phospholipase A(2) (PLA(2)) (AACOCF(3)), secretory PLA(2) (SB-203347), protein kinase (PK) (staurosporine), PKC (GF-109203; BIM), and serine protease (Pefabloc SC), antagonists of thromboxane prostanoid (TP) receptor (R) (SQ-29548), platelet activating factor (PAF) R (BN-52021), leukotriene B(4) R (SC-41930), serotonin R (cyproheptadine), fibronectin/fibrinogen R (RGDS), and finally, creatine phosphate/creatine phosphokinase (CP/CPK) which removes ADP. Whereas when added alone neither of these agents significantly inhibited LPS-induced TF or TNF-alpha, when presented as a reference cocktail comprising all the agents, TF activity and TNF-alpha were reduced by 77% and 49%, respectively. By subsequently testing a series of incomplete inhibitory cocktails equal to the reference except for deleted single agents or combinations of two or three active agents, the inhibitory effect of the reference cocktail could be shown to depend on the presence of the protease inhibitor and the thromboxane A(2) and PAF antagonists.
Eilertsen
Karl-Erik
KE
Department of Biochemistry, Institute of Medical Biology, Faculty of Medicine, University of Tromsø, Tromsø, Norway. Karl-Erik.Eilertsen@fagmed.uit.no
Østerud
Bjarne
B
D001095:Arachidonic Acids / Q000494:pharmacology; D001578:Benzopyrans / Q000494:pharmacology; D019086:Bridged Bicyclo Compounds, Heterocyclic; D003402:Creatine Kinase / Q000494:pharmacology; D003533:Cyproheptadine / Q000494:pharmacology; D004224:Diterpenes; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli / Q000276:immunology*; D005231:Fatty Acids, Unsaturated; D046934:Ginkgolides; D006801:Humans; D006834:Hydrazines / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D007783:Lactones / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008301:Maleimides / Q000494:pharmacology; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism*; D009842:Oligopeptides / Q000494:pharmacology; D010980:Platelet Membrane Glycoproteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled; D017482:Receptors, Thromboxane / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013449:Sulfonamides / Q000494:pharmacology; D013450:Sulfones / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D001095:Arachidonic Acids; D001578:Benzopyrans; D019086:Bridged Bicyclo Compounds, Heterocyclic; D004224:Diterpenes; D004791:Enzyme Inhibitors; D005231:Fatty Acids, Unsaturated; D046934:Ginkgolides; D006834:Hydrazines; D007211:Indoles; D007783:Lactones; D008070:Lipopolysaccharides; D008301:Maleimides; D009842:Oligopeptides; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D017482:Receptors, Thromboxane; C095801:SB 203347; D013449:Sulfonamides; D013450:Sulfones; D014409:Tumor Necrosis Factor-alpha; C039351:platelet activating factor receptor; C081565:arachidonyltrifluoromethane; C059950:SC 41930; D003533:Cyproheptadine; C002010:4-(2-aminoethyl)benzenesulfonylfluoride; C045749:SQ 29548; C046617:arginyl-glycyl-aspartyl-serine; C045856:ginkgolide B; D003402:Creatine Kinase; C070515:bisindolylmaleimide I