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Gamma-Linolenic Acid Inhibits Inflammatory Responses by Regulating NF-κB and AP-1 Activation in Lipopolysaccharide-Induced RAW 264.7 Macrophages

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Abstract

Gamma linolenic acid (GLA) is a member of the n-6 family of polyunsaturated fatty acids and can be synthesized from linoleic acid (LA) by the enzyme delta-6-desaturase. The therapeutic values of GLA supplementation have been documented, but the molecular mechanism behind the action of GLA in health benefits is not clear. In this study, we assessed the effect of GLA with that of LA on lipopolysaccharide (LPS)-induced inflammatory responses and further explored the molecular mechanism underlying the pharmacological properties of GLA in mouse RAW 264.7 macrophages. GLA significantly inhibited LPS-induced protein expression of inducible nitric oxide synthase, pro-interleukin-1β, and cyclooxygenase-2 as well as nitric oxide production and the intracellular glutathione level. LA was less potent than GLA in inhibiting LPS-induced inflammatory mediators. Both GLA and LA treatments dramatically inhibited LPS-induced IκB-α degradation, IκB-α phosphorylation, and nuclear p65 protein expression. Moreover, LPS-induced nuclear factor-κB (NF-κB) and activator protein-1 (AP-1) nuclear protein–DNA binding affinity and reporter gene activity were significantly decreased by LA and GLA. Exogenous addition of GLA but not LA significantly reduced LPS-induced expression of phosphorylated extracellular signal-regulated kinase (ERK) 1/2 and c-Jun N-terminal kinase (JNK)-1. Our data suggest that GLA inhibits inflammatory responses through inactivation of NF-κB and AP-1 by suppressed oxidative stress and signal transduction pathway of ERK and JNK in LPS-induced RAW 264.7 macrophages.

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Abbreviations

AP-1:

activator protein-1

COX-2:

cyclooxygenase-2

EMSA:

electrophoretic mobility shift assay

ERK1/2:

extracellular signal-regulated kinase1/2

GAPDH:

glyceraldehydes-3-phosphate dehydrogenase

GLA:

gamma-linolenic acid

GSH:

glutathione

IL-1β:

interleukin-1β

iNOS:

inducible form of NOS

IS:

internal standard

JNK:

c-Jun NH2-terminal kinase

LA:

linoleic acid

LPS:

lipopolysaccharide

MAPK:

mitogen-activated protein kinase

MTT:

3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide

NF-κB:

nuclear factor-κB

NO:

nitric oxide

NOS:

nitric oxide synthase

PGE2 :

prostaglandin E2

PMSF:

phenylmethylsulfonyl fluoride

rcRNA:

recombinant RNA

RT-PCR:

reverse transcriptase polymerase chain reaction

SEAP:

secretory alkaline phosphatase

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Acknowledgments

This research was funded by the National Science Council, Republic of China, under Grant NSC 95-2320-B-040-034 and by Chung Shan Medical University, under Grant CSMU 94-OM-B-003.

Author information

Authors and Affiliations

  1. Department of Neurology, Changhua Christian Hospital, Changhua City, Changhua County, Taiwan

    Cheng-Sue Chang

  2. Department of Allergy, Chung Shan Medical University Hospital, Taichung, Taiwan

    Hai-Lun Sun

  3. School of Medicine, Chung Shan Medical University, Taichung, Taiwan

    Hai-Lun Sun

  4. Department of Nutrition, China Medical University, Taichung, Taiwan

    Chong-Kuei Lii & Haw-Wen Chen

  5. Department of Nutrition, Chung Shan Medical University, No. 110, Sec. 1, Chien-Kuo North Road, Taichung, 40203, Taiwan

    Pei-Yin Chen & Kai-Li Liu

  6. Department of Dietitian, Chung Shan Medical University Hospital, Taichung, Taiwan

    Kai-Li Liu

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  1. Cheng-Sue Chang
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Correspondence to Kai-Li Liu.

Additional information

Cheng-Shu Chang and HaiLun Sun contributed equally to this work.

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Chang, CS., Sun, HL., Lii, CK. et al. Gamma-Linolenic Acid Inhibits Inflammatory Responses by Regulating NF-κB and AP-1 Activation in Lipopolysaccharide-Induced RAW 264.7 Macrophages. Inflammation 33, 46–57 (2010). https://doi.org/10.1007/s10753-009-9157-8

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  • DOI: https://doi.org/10.1007/s10753-009-9157-8

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