Human Neutrophils Convert the Sebum-derived Polyunsaturated Fatty Acid Sebaleic Acid to a Potent Granulocyte Chemoattractant

Orval A Mamer,Sashikala Sivendran,Sylvie Gravel,Gue Jae Lee,Chantal Cossette,Jaganmohan R Anumolu,Pranav Patel,William S Powell,François D Graham,Joshua Rokach,Alain Lesimple

doi:10.1074/jbc.m709531200

Orval A Mamer, Sashikala Sivendran + Show 9 more

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https://doi.org/10.1074/jbc.m709531200

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Abstract

Sebaleic acid (5,8-octadecadienoic acid) is the major polyunsaturated fatty acid in human sebum and skin surface lipids. The objective of the present study was to investigate the metabolism of this fatty acid by human neutrophils and to determine whether its metabolites are biologically active. Neutrophils converted sebaleic acid to four major products, which were identified by their chromatographic properties, UV absorbance, and mass spectra as 5-hydroxy-(6E,8Z)-octadecadienoic acid (5-HODE), 5-oxo-(6E,8Z)-octadecadienoic acid (5-oxo-ODE), 5S,18-dihydroxy-(6E,8Z)-octadecadienoic acid, and 5-oxo-18-hydroxy-(6E,8Z)-octadecadienoic acid. The identities of these metabolites were confirmed by comparison of their properties with those of authentic chemically synthesized standards. Both neutrophils and human keratinocytes converted 5-HODE to 5-oxo-ODE. This reaction was stimulated in neutrophils by phorbol myristate acetate and in keratinocytes by oxidative stress (t-butyl-hydroperoxide). Both treatments dramatically elevated intracellular levels of NADP(+), the cofactor required by 5-hydroxyeicosanoid dehydrogenase. In keratinocytes, this was accompanied by a rapid increase in intracellular GSSG levels, consistent with the involvement of glutathione peroxidase. 5-Oxo-ODE stimulated calcium mobilization in human neutrophils and induced desensitization to 5-oxo-6,8,11,14-eicosatetraenoic acid but not leukotriene B(4), indicating that this effect was mediated by the OXE receptor. 5-Oxo-ODE and its 8-trans isomer were equipotent with 5-oxo-6,8,11,14-eicosatetraenoic acid in stimulating actin polymerization and chemotaxis in human neutrophils, whereas 5-HODE, 5-oxo-18-hydroxy-(6E,8Z)-octadecadienoic acid, and 5S,18-dihydroxy-(6E,8Z)-octadecadienoic acid were much less active. We conclude that neutrophil 5-lipoxygenase converts sebaleic acid to 5-HODE, which can be further metabolized to 5-oxo-ODE by 5-hydroxyeicosanoid dehydrogenase in neutrophils and keratinocytes. Because of its chemoattractant properties, sebum-derived 5-oxo-ODE could be involved in neutrophil infiltration in inflammatory skin diseases.

Highlights

One possible route for the oxidative metabolism of sebaleic acid is the 5-lipoxygenase (5-LO) pathway. 5-LO converts the ␻6 polyunsaturated fatty acids (PUFA) arachidonic acid (5,8,11,14-eicosatetraenoic acid) to
Since sebaleic acid is a ⌬5,8-PUFA like arachidonic acid, we hypothesized that it might be a substrate for 5-LO and could possibly be converted to an analogous 5-hydroxy metabolite, further conversion to leukotrienes would not be possible due to the lack of a ⌬11-double bond
There were peaks corresponding to metabolites of endogenous arachidonic acid, which were present when neutrophils were incubated with A23187 and PMA in the absence of sebaleic acid

Summary

Introduction

One possible route for the oxidative metabolism of sebaleic acid is the 5-lipoxygenase (5-LO) pathway. 5-LO converts the ␻6 PUFA arachidonic acid (5,8,11,14-eicosatetraenoic acid) to. Metabolism of Sebaleic Acid to the Chemoattractant 5-Oxo-ODE (Sigma) [17]. 5-hydroperoxy-6,8,11,14-eicosatetraenoic acid and leukot- decadienoic acid (8-trans-5-oxo-ODE; Fig. 1E) were prepared rienes [7]. 5R-HODE converted to leukotriene B4 (LTB4), a potent neutrophil chemoattractant, and 5-hydroxy-6,8,11,14-eicosatetraenoic acid (5-HETE), respectively [8]. 5-HETE has only weak biological activities on neutrophils and eosinophils, it is oxidized to the potent granulocyte chemoattractant 5-oxo-6,8,11,14-eicosatetraenoic acid (5-oxoETE) by the NADPϩ-dependent enzyme 5-hydroxyeicosanoid dehydrogenase (5-HEDH) [9]. The objective of the current study was to determine whether sebaleic acid is a substrate for the 5-LO pathway and whether it can be converted to biologically active proinflammatory products that could potentially be involved in attracting granulocytes to the skin

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