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Open Access

Modulation of oxidative stress and TNFα secretion by peritoneal macrophages of arthritic rats fed with a flavonoid-enriched diet

  • S Ramos-Romero1,
  • F J Pérez-Cano1,
  • C Ramírez-Santana1,
  • C Castellote1,
  • A Franch1 and
  • M Castell1
Journal of Translational Medicine20108(Suppl 1):P24

https://doi.org/10.1186/1479-5876-8-S1-P24

Published: 25 November 2010

Keywords

FlavonoidPeritoneal MacrophageDichlorofluoresceinGriess ReactionDichlorofluorescein Diacetate

Introduction

The inflammatory response increases the oxidative stress and cytokine production, such as TNFα, in macrophages. Flavonoids, present in vegetables, have potential antioxidant and anti-inflammatory properties. Previous studies have shown that a flavonoid-enriched cocoa extract added to macrophages in vitro decreases the secretion of NO and TNFα [1].

Aim

The objective of the present study was to ascertain the production of oxidant products and a pro-inflammatory cytokine (TNFα) by peritoneal macrophages obtained from arthritic rats after a long-term cocoa diet.

Methods

Female LOU rats were fed with a 10% cocoa diet or standard chow. After 2 weeks of diet, collagen-induced arthritis (CIA) was induced in part of animals from each group. One month later, peritoneal macrophages were obtained by injecting ice-cold sterile PBS into the peritoneal cavity. Cells were cultured and stimulated by addition of 1 µg/mL LPS. TNFα secretion was quantified by ELISA in 24 h supernatants. In the same samples, NO2- concentration (stable end product of NO) was measured by a modification of Griess reaction. To determine intracellular ROS production, macrophages were incubated with reduced 2’,7’-dichlorofluorescein diacetate (H2DCF-DA) for 30 min at 37 ºC. Fluorescence of DCF oxidized by ROS was measured by fluorometry every 30 min up to 2 h.

Results

Macrophages from CIA animals fed cocoa decreased TNFα and NO production by ~57% and ~36% with respect to those from CIA reference group under LPS-stimulation conditions (P<0.05). ROS production by macrophages from CIA animals fed standard diet was higher with respect to healthy reference animals (P<0.05). Cocoa diet avoided ROS secretion increase in CIA animals (P<0.05), raising similar values to healthy animals.

Conclusion

A long-term cocoa diet reduced the production of oxidants and TNFα in peritoneal macrophages obtained from collagen-induced arthritic rats.

Authors’ Affiliations

(1)
Dept. of Physiology, Faculty of Pharmacy, University of Barcelona, Spain

References

  1. Ramiro E, Franch A, Castellote C, Pérez-Cano FJ, Permanyer J, Izquierdo-Pulido M, Castell M: Flavonoids from Theobroma cacao down-regulate inflammatory mediators. J Agric Food Chem. 2005, 53: 8506-8511. 10.1021/jf0511042.View ArticlePubMedGoogle Scholar

Copyright

© Castell et al; licensee BioMed Central Ltd. 2010

This article is published under license to BioMed Central Ltd.

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