Abstract
Researchers in the eicosanoid field have frequently applied whole blood assays (1–3) for evaluation of enzymic inhibitors in preclinical and clinical studies. In numerous investigations of antiplatelet agents, the capacity of blood to generate thromboxane A2 (TXA2) in response to an agonist has been assessed, and similar assays figure importantly in the current clinical trials (4) of cyclooxygenase-2 (COX-2) inhibitors. Early work by Sun (5), carried out with human platelet microsomes, established the reaction conditions for studies of thromboxane synthase, and demonstrated that thromboxane synthase is not inhibited by non steroidal anti-inflammatory agents, such as aspirin. It now seems clear that other blood elements, notably monocytes, also contain thromboxane synthase. When the objective is to manipulate, pharmacologically, arachidonic acid (AA) metabolism, or to track routes of transcellular metabolism of AA (discussed in Chapter 16), convenient assays are often set up to measure stable products, such as 6-keto-PGF1α and TXB2; hydrolysis products of prostacyclin (PGI2) and TXA2, respectively. Depending on the specific purpose, different agonists (e.g., thrombin, collagen, endotoxin) may be used to elicit formation of eicosanoids by blood elements.
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References
Spaethe, R. (1984) Haemostasis: Physiology, Pathophysiology, Diagnostics. AHS/Deutschland GmmH, Munchen, Germany.
Stier, C. T., Jr. and Spokas, E. G. (1988) The role of thromboxane A2 in hypertension. Cardiovasc. Rev. Rep. 9(8), 32–38.
Sinzinger, H., O’Grady, J., Demers, L. M., Granstrom, E., Kumlin, M., Nell, A., Peskar, B. A., Salmon, J. A., and Westlund, P. (1990) Thromboxane in cardiovascular disease. Eicosanoids 3(1), 59–64.
Nies, A. S. (1995) The Eicosanoids: a personal journal of discovery. Lecture presented on receiving the 1995 Harry Gold Award. Pharmacologist 37(3), 92–101.
Sun, F. F. (1977) Biosynthesis of thromboxanes in human platelets. I. Characterization and assay of thromboxane synthetase. Biochem. Biophys. Res. Commun. 74(4), 1432–1440.
Mehta, J., Mehta, P., Lawson, D. L., Ostrowski, N., and Brigmon, L. (1985) Influence of selective thromboxane synthetase blocker CGS-13080 on thromboxane and prostacyclin synthesis in whole blood: evidence for synthesis of prostacyclin by leukocytes from platelet-derived endoperoxides. J. Lab. Clin. Med. 106, 246–252.
Simpson, P. J., Smith, C. B., Jr., Rosenthal, G., and Lucchesi, B. R. (1986) Reduction in the incidence of thrombosis by the thromboxane synthetase inhibitor CGS 13080 in a canine model of coronary artery injury. J. Pharmacol. Exper. Ther. 238(2), 497–501.
McCullagh, K. G., Tuffin, D. P., Honey, A., Lad, N., Manley, P., Myers, P., Porter, R., Wade P., and Booth, R. (1986) SC 41156: A novel inhibitor of human thromboxane synthase. Abstract presented at the 6th International Conference on Prostglandins and Related Compounds, Florence, Italy, June 3–6, 1986.
Manley, P.W, Tuffin, D. P., Allanson, N. M., Buckle, P. E., Lad, N., Lai, S. M. F., Lunt, D. O., Porter, R. A., and Wade, P. J. (1987) Thromboxane synthase inhibitors. Synthesis and pharmacological activity of (R)-, (S)-, and (+)-2,2-dimethyl-6-[2-(1H-imidazol-1-yl)-1-[[94-methoxyphenyl0-methoxy]methyl]ethoxy] hexanoic acids. J. Med. Chem. 30, 1812–1818.
Manley, P.W, Allanson, N. G., Booth, R. F. G., Buckle, P. E., Kuzniar, E. J., Lad, N., Lai, S. M. F., Lunt, D. O., and Tuffin, D. P. (1987) Structure-activity relationships in an imidazole-based series of thromboxane synthase inhibitors. J. Med. Chem. 30, 1588–1595.
Stier, C. T., Jr. and Itskovitz, H. D. (1988) Thromboxane A2 and the development of hypertension in spontaneously hypertensive rats. Eur. J. Pharmacol. 146, 129.
Defreyn, G., Deckmyn, H., and Vermylen, J. (1982) A thromboxane synthetase inhibitor reorients endoperoxide metabolism in whole blood towards prostacyclin and prostaglandin E2. Thromb. Res. 26, 389–400.
Spokas, E. G., Suleymanov, O. D., Bittner, S. E., Campion, J. G., Gorczynski, R. J., Lenaers, A., and Walsh, G. M. (1987) Cardiovascular effects of chronic high-dose atriopeptin III infusion in normotensive rats. Toxicol. Appl. Pharmacol. 91, 305–314.
Sinzinger, H., Reiter, S., and Peskar, B. A. (1984) Removal, preparation and storage of human plasma for radioimmunological detection of prostaglandins, in Prostaglandins and other Eicosanoids in the Cardiovascular System (Schror, K., ed.), Karger, Basel, Switzerland, pp. 62–67.
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Spokas, E.G., Nicholson, N.S., Suleymanov, O.D., Walsh, G.M., Tuffin, D. (1999). Whole Blood Assays for Evaluation of Thromboxane Synthase Inhibition. In: Lianos, E.A. (eds) Eicosanoid Protocols. Methods in Molecular Biology, vol 120. Humana Press. https://doi.org/10.1385/1-59259-263-5:249
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DOI: https://doi.org/10.1385/1-59259-263-5:249
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