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Protectin DX, a Double Lipoxygenase Product of DHA, Inhibits Both ROS Production in Human Neutrophils and Cyclooxygenase Activities

  • Original Article
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Lipids

Abstract

Neutrophils play a major role in inflammation by releasing large amounts of reactive oxygen species (ROS) produced by NADPH oxidase (NOX) and myeloperoxidase (MPO). This ROS overproduction is mediated by phosphorylation of the NOX subunits in an uncontrolled manner. Therefore, targeting neutrophil subunits would represent a promising strategy to moderate NOX activity, lower ROS, and other inflammatory agents, such as cytokines and leukotrienes, produced by neutrophils. For this purpose, we investigated the effects of protectin DX (PDX)—a docosahexaenoic acid di-hydroxylated product which inhibits blood platelet aggregation—on neutrophil activation in vitro. We found that PDX decreases ROS production, inhibits NOX activation and MPO release from neutrophils. We also confirm, that PDX is an anti-aggregatory and anti-inflammatory agent by inhibiting both cyclooxygenase-1 and -2 (COX-1 and COX-2, E.C. 1.14.99.1) as well as COX-2 in lipopolysaccharides-treated human neutrophils. However, PDX has no effect on the 5-lipoxygenase pathway that produces the chemotactic agent leukotriene B4 (LTB4). Taken together, our results suggest that PDX could be a protective agent against neutrophil invasion in chronic inflammatory diseases.

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Abbreviations

ARA:

Arachidonic acid

BSTFA:

N,O-Bis(trimethylsilyl)-trifluoroacetamide

CLA:

Conjugated linoleic acid

ClnA:

Conjugated linolenic acid

COX:

Cyclooxygenase

DHA:

Docosahexaenoic acid

ECL:

Enhanced chemiluminescence

EPA:

Eicosapentaenoic acid

5-HETE:

5-hydroxy-6E,8Z,11Z,14Z-eicosatetraenoic acid

fMLF:

Formyl-methionyl-leucylphenylalanine

GM-CSF:

Granulocyte macrophage-colony stimulating factor

HBSS:

Hank’s balanced salt solution

HOCl:

Hypochloric acid

HRP:

Horseradish peroxidase

IBD:

Inflammatory bowel disease

IL:

Interleukin

LA:

Linoleic acid

LPS:

Lipopolysaccharides

LTB4 :

Leukotriene B4

MPO:

Myeloperoxidase

NOX:

NADPH oxidase

PAF:

Platelet-activating factor

PAGE:

Polyacrylamide gel electrophoresis

PBS:

Phosphate-buffered saline

PDX:

Protectin DX

PG:

Prostaglandin

Phox:

Phagocyte oxidase

PMA:

Phorbol myristate acetate

PMN:

Polymorphonuclear neutrophil

PUFA:

Polyunsaturated fatty acids

ROS:

Reactive oxygen species

RP-HPLC:

Reverse phase high performance liquid chromatography

sLOX:

Soybean lipoxygenase

TLC:

Thin-layer chromatography

TMB:

Tetramethyl benzidine

TNBS:

Trinitrobenzene sulfonic acid

TNF-α:

Tumor necrosis factor-α

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Acknowledgments

This study was supported by Inserm and the French Ministry of Education and Research. Miao Liu was received a grant from the China Scholarship Council.

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Authors and Affiliations

  1. UMR 1060 Inserm (CarMeN), IMBL/INSA-Lyon, Université de Lyon, 69621, Villeurbanne, France

    Miao Liu, Michel Lagarde & Michel Guichardant

  2. Inserm, UMR 773, CRB3, Faculté de Médecine, Université Paris 7 Denis Diderot, Site Bichat, 75018, Paris, France

    Tarek Boussetta, Karama Makni-Maalej, Michèle Fay, Fathi Driss & Jamel El-Benna

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  1. Miao Liu
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  2. Tarek Boussetta
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Correspondence to Michel Guichardant.

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M. Liu, T. Boussetta, K. Makni-Maalej contributed equally to this work.

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Liu, M., Boussetta, T., Makni-Maalej, K. et al. Protectin DX, a Double Lipoxygenase Product of DHA, Inhibits Both ROS Production in Human Neutrophils and Cyclooxygenase Activities. Lipids 49, 49–57 (2014). https://doi.org/10.1007/s11745-013-3863-6

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  • DOI: https://doi.org/10.1007/s11745-013-3863-6

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