Metabolism of eicosapentaenoic and docosahexaenoic acids by recombinant human cytochromes P450

doi:10.1016/j.abb.2008.01.002

Archives of Biochemistry and Biophysics

Volume 471, Issue 2, 15 March 2008, Pages 116-125

https://doi.org/10.1016/j.abb.2008.01.002 Get rights and content

Abstract

Epoxidation and hydroxylation of arachidonic acid (AA) are both catalyzed by cytochromes P450s (CYPs). The oxidized metabolites are known to be involved in the regulation of vascular tone and renal function. By using a panel of 15 human recombinant CYPs, this study demonstrates that other polyunsaturated long-chain fatty acids (PUFA-LC), especially the ω3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are also epoxidised. The regioselectivity of epoxidation of four PUFA-LC by CYPs was investigated. Among the several CYPs tested, CYP2C9/2C19 and 1A2 were the most efficient in EPA and DHA epoxidations. It ensued that 10 μM of these two ω3 fatty acids decreased by more than 80% and 60%, respectively, the formation by CYP2C9 of AA-epoxidised derivatives. These findings suggest that some physiological effects of ω3 fatty acids may be due to a shift in the generation of active epoxidised metabolites of AA through CYP-mediated catalysis.

Section snippets

Chemicals

All chemicals of analytical grade were from Sigma-Aldrich (L’Isle d’Abeau, France). HPLC-purity grade solvents were from Carlo Erba (Val de Reuil, France). The counter-ion for HPLC, n-dibutylamine (DBA) in its acetate concentrated form as well as m-chloroperbenzoic acid (m-CPBA) were both from Fluka (Buchs, Switzerland). The PUFAs (5,8,11-ETA, AA, EPA and DHA) and their epoxides (5,6-EET, 8,9-EET, 11,12-EET, 14,15-EET, 14,15-EETeTr, 16,17-EDP and 19,20-EDP) were from Cayman Chemicals (Spi-Bio,

Identification of metabolites

The presence of several (3–6) double bonds in PUFAs skeleton leads to their transformation into numerous epoxide regioisomers. This consideration has driven our group to develop a reversed-phase ion-pair HPLC method to separate, in a single run, all of the monoepoxides, ω- and (ω-1)-hydroxylated derivatives issued from PUFAs [22]. After separation by HPLC, under ion-pair conditions, PUFAs metabolites were identified by LC-MS with APCI source in negative mode. The monooxidized compounds were

Discussion

Cytochromes P450 are known to metabolize AA to HETEs and EETs [1], [2], [3]. Biological activities of EETs and the opposing actions of 20-HETE within the vasculature are now well established [2], [12]. By contrast, little is known about the CYP-dependent metabolism of PUFAs other than AA, as about the metabolic interaction between AA and ω3 PUFAs. Using the reversed-phase ion-pair HPLC method developed in our laboratory [22], we investigated the formation of epoxidized and hydoxylated

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¹: M.F. was supported by a fellowship from the Ministère de l’Enseignement Supérieur et de la Recherche.

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Metabolism of eicosapentaenoic and docosahexaenoic acids by recombinant human cytochromes P450

Abstract

Section snippets

Chemicals

Identification of metabolites

Discussion

Prog. Lipid Res.

J. Biol. Chem.

Arch. Biochem. Biophys.

Prostaglandins Other Lipid Mediat.

Biochem. Pharmacol.

Biochem. Biophys. Res. Commun.

Prostaglandins Leukot. Essent. Fatty Acids

Biochim. Biophys. Acta

J. Biol. Chem.

Am. J. Clin. Nutr.

J. Chromatogr. A

J. Lipid Res.

Anal. Biochem.

Anal. Biochem.

J. Lipid Res.

Arch. Biochem. Biophys.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Arch. Biochem. Biophys.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

Drug Metab. Pharmacokinet.

Arch. Biochem. Biophys.

Biochem. Pharmacol.

J. Biol. Chem.