Lipids

, Volume 49, Issue 1, pp 49–57 | Cite as

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

  • Miao Liu
  • Tarek Boussetta
  • Karama Makni-Maalej
  • Michèle Fay
  • Fathi Driss
  • Jamel El-Benna
  • Michel Lagarde
  • Michel Guichardant
Original Article

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.

Keywords

Human neutrophils Inflammation Protectin DX NADPH oxidase ROS-derived oxidative stress 

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

Notes

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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Copyright information

© AOCS 2013

Authors and Affiliations

  • Miao Liu
    • 1
  • Tarek Boussetta
    • 2
  • Karama Makni-Maalej
    • 2
  • Michèle Fay
    • 2
  • Fathi Driss
    • 2
  • Jamel El-Benna
    • 2
  • Michel Lagarde
    • 1
  • Michel Guichardant
    • 1
  1. 1.UMR 1060 Inserm (CarMeN), IMBL/INSA-LyonUniversité de LyonVilleurbanneFrance
  2. 2.Inserm, UMR 773, CRB3, Faculté de MédecineUniversité Paris 7 Denis DiderotParisFrance

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