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J Ethnopharmacol. 2013 Jan 9;145(1):42-9. doi: 10.1016/j.jep.2012.10.031. Epub 2012 Nov 6.

Mechanisms of anti-inflammatory property of Anacardium occidentale stem bark: inhibition of NF-κB and MAPK signalling in the microglia.

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1
Division of Pharmacy and Pharmaceutical Science, School of Applied Sciences, University of Huddersfield, Queensgate, Huddersfield HD1 3DH, United Kingdom. o.a.olajide@hud.ac.uk

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE:

Anacardium occidentale is used in traditional African medicine for the treatment of arthritis, fever, aches, pains, and inflammation of the extremities.

AIM OF THE STUDY:

In this study, we investigated the molecular mechanisms responsible for anti-inflammatory effects of a stem bark extract of A. occidentale (ANE) in LPS-stimulated microglia.

MATERIALS AND METHODS:

Nitric oxide (NO), prostaglandin E(2) and cytokine (TNFα and IL-6) production were evaluated in supernatants from LPS-stimulated BV-2 cells. Cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS) and microsomal prostaglandin E2 synthase (mPGES-1) protein expressions in rat primary microglia were measured using western blot. The effects of ANE on NF-κB activation and nuclear translocation were evaluated in the luciferase reporter gene assay and ELISA, while ability of ANE to influence IκB phosphorylation was determined using ELISA specific for phospho-IκB. The involvement of MAPK phosphorylation in the anti-inflammatory actions of ANE was evaluated using specific ELISA for phospho-p38, phospho-p42/44 and phospho-JNK. The MTT assay was used to determine the effect of ANE on BV-2 microglia viability.

RESULTS:

ANE (25-100 μg/ml) produced significant (p<0.05) reduction in the production of NO, PGE(2), TNFα and IL-6 in BV-2 microglia stimulated with LPS for 24h. Pre-treatment with ANE caused a significant (p<0.05) inhibition of COX-2, iNOS and mPGES-1 protein expressions in the rat primary microglia. Further experiments showed that ANE inhibited COX-2 and iNOS protein expression via IκB-mediated nuclear translocation and transactivation of NF-κB. Our studies also revealed that ANE produced significant (p<0.05) and dose-dependent inhibition of p38, p42/44 and JNK MAPK phosphorylation in LPS-activated BV-2 microglia.

CONCLUSIONS:

We conclude that ANE has an anti-inflammatory property related to inhibition of inflammation-associated cytokine production as well as iNOS and COX-2 gene expression by blocking NF-κB and MAPK pathways in the microglia. It is also suggested that mPGES-1 inhibition contributes to the effect of ANE on PGE(2) production in the microglia.

PMID:
23142196
DOI:
10.1016/j.jep.2012.10.031
[Indexed for MEDLINE]
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