Abstract
The assessment of prostanoids (TXB2, 6-keto-PGF1α, PGD2 and PGE2) generated in isolated or cultured cells in vitro can be directly assessed in media without extraction mainly using immunoassays, i.e. radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), or enzyme immunoassay (EIA). However, the immunoassays have to have some fundamental requisites, such as the use of highly specific and sensitive antibodies. Importantly, the results obtained with immunoassays should be validated by comparison with gas chromatography (GC), mass spectrometry (MS), or liquid chromatography (LC)–MS. The measurement of prostanoid generation in vivo can be performed by the evaluation of prostaglandin breakdown products in urine using LC–MS, GC–MS, or validated immunoassays after extraction and rigorous sample purification. The measurement of F2-isoprostanes (F2-iPs), containing F-type ring analogous to PGF2α, provides a reliable tool for identifying populations with enhanced rates of lipid peroxidation. Levels of 8- iso-PGF2α (also referred as iPF2α-III) in plasma or urine are assessed by GC/MS or validated immunoassays.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Reid G, Wielinga P, Zelcer N, van der Heijden I, Kuil A, de Haas M, Wijnholds J, Borst P (2003) The human multidrug resistance protein MRP4 functions as a prostaglandin efflux transporter and is inhibited by nonsteroidal antiinflammatory drugs. Proc Natl Acad Sci U S A 100:9108–9110
FitzGerald GA (2003) COX-2 and beyond: approaches to prostaglandin inhibition in human disease. Nat Rev Drug Discov 11:879–890
Simmons DL, Botting RM, Hla T (2004) Cyclooxygenase isozymes: the biology of prostaglandin synthesis and inhibition. Pharmacol Rev 56:387–437
Samuelsson B, Morgenstern R, Jakobsson PJ (2007) Membrane prostaglandin E synthase-1: a novel therapeutic target. Pharmacol Rev 59:207–224
Kudo I, Murakami M (2005) Prostaglandin E synthase, a terminal enzyme for prostaglandin E2 biosynthesis. J Biochem Mol Biol 38:633–638
Urade Y, Eguchi N (2002) Lipocalin-type and hematopoietic prostaglandin D synthases as a novel example of functional convergence. Prostaglandins Other Lipid Mediat 68–69:375–382
Wu KK, Liou JY (2005) Cellular and molecular biology of prostacyclin synthase. Biochem Biophys Res Commun 338:45–52
Ueno N, Takegoshi Y, Kamei D, Kudo I, Murakami M (2005) Coupling between cyclooxygenases and terminal prostanoid synthases. Biochem Biophys Res Commun 338:70–76
Komoto J, Yamada T, Watanabe K, Woodward DF, Takusagawa F (2006) Prostaglandin F2alpha formation from prostaglandin H2 by prostaglandin F synthase (PGFS): crystal structure of PGFS containing bimatoprost. Biochemistry 45:1987–1996
Haurand M, Ullrich V (1985) Isolation and characterization of thromboxane synthase from human platelets as a cytochrome P-450 enzyme. J Biol Chem 260:15059–15067
Kang YJ, Mbonye UR, DeLong CJ, Wada M, Smith WL (2007) Regulation of intracellular cyclooxygenase levels by gene transcription and protein degradation. Prog Lipid Res 46:108–125
Topper JN, Cai J, Falb D, Gimbrone MA Jr (1996) Identification of vascular endothelial genes differentially responsive to fluid mechanical stimuli: cyclooxygenase-2, manganese superoxide dismutase, and endothelial cell nitric oxide synthase are selectively up-regulated by steady laminar shear stress. Proc Natl Acad Sci U S A 93:10417–10422
FitzGerald GA, Pedersen AK, Patrono C (1983) Analysis of prostacyclin and thromboxane biosynthesis in cardiovascular disease. Circulation 67:1174–1177
Ciabattoni G, Maclouf J, Catella F, FitzGerald GA, Patrono C (1987) Radioimmunoassay of 11-dehydrothromboxane B2 in human plasma and urine. Biochim Biophys Acta 918:293–297
FitzGerald GA, Brash AR, Falardeau P, Oates JA (1981) Estimated rate of prostacyclin secretion into the circulation of normal man. J Clin Invest 68:1272–1276
Murphey LJ, Williams MK, Sanchez SC, Byrne LM, Csiki I, Oates JA, Johnson DH, Morrow JD (2004) Quantification of the major urinary metabolite of PGE2 by a liquid chromatographic/mass spectrometric assay: determination of cyclooxygenase-specific PGE2 synthesis in healthy humans and those with lung cancer. Anal Biochem 334:266–275
Song WL, Wang M, Ricciotti E, Fries S, Yu Y, Grosser T, Reilly M, Lawson JA, FitzGerald GA (2008) Tetranor PGDM, an abundant urinary metabolite reflects biosynthesis of prostaglandin D2 in mice and humans. J Biol Chem 283:1179–1188
Granström E, Samuelsson B (1971) On the metabolism of prostaglandin F2a in female subjects. II. Structures of six metabolites. J Biol Chem 246:7470–7485
Patrono C, Ciabattoni G, Pinca E, Pugliese F, Castrucci G, De Salvo A, Satta MA, Peskar BA (1980) Low dose aspirin and inhibition of thromboxane B2 production in healthy subjects. Thromb Res 17:317–327
Patrignani P, Panara MR, Greco A, Fusco O, Natoli C, Iacobelli S, Cipollone F, Ganci A, Créminon C, Maclouf J, Patrono C (1994) Biochemical and pharmacological characterization of the cyclooxygenase activity of human blood prostaglandin endoperoxide synthases. J Pharmacol Exp Ther 271:1705–1712
Capone ML, Tacconelli S, Sciulli MG, Grana M, Ricciotti E, Minuz P, Di Gregorio P, Merciaro G, Patrono C, Patrignani P (2004) Clinical pharmacology of platelet, monocyte, and vascular cyclooxygenase inhibition by naproxen and low-dose aspirin in healthy subjects. Circulation 109:1468–1471
Roberts LJ 2nd, Morrow JD (2002) Products of the isoprostane pathway: unique bioactive compounds and markers of lipid peroxidation. Cell Mol Life Sci 59:808–820
Patrignani P, Panara MR, Tacconelli S, Seta F, Bucciarelli T, Ciabattoni G, Alessandrini P, Mezzetti A, Santini G, Sciulli MG, Cipollone F, Davì G, Gallina P, Bon GB, Patrono C (2000) Effects of vitamin E supplementation on F2-isoprostane and thromboxane biosynthesis in healthy cigarette smokers. Circulation 102:539–545
Rokach J, Khanapure SP, Hwang SW, Adiyaman M, Lawson JA, FitzGerald GA (1997) Nomenclature of isoprostanes: a proposal. Prostaglandins 54:853–873
Wang Z, Ciabattoni G, Créminon C, Lawson J, Fitzgerald GA, Patrono C, Maclouf J (1995) Immunological characterization of urinary 8-epi-prostaglandin F2α excretion in man. J Pharmacol Exp Ther 275:94–100
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Patrignani, P. (2010). Extraction and Measurement of Prostanoids and Isoprostanes: Introduction to Part II. In: Ayoub, S., Flower, R., Seed, M. (eds) Cyclooxygenases. Methods in Molecular Biology, vol 644. Humana Press. https://doi.org/10.1007/978-1-59745-364-6_12
Download citation
DOI: https://doi.org/10.1007/978-1-59745-364-6_12
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-58829-953-6
Online ISBN: 978-1-59745-364-6
eBook Packages: Springer Protocols