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Eur J Nutr (2009) 48:5461DOI 10.1007/s00394-008-0761-4
ORIGINAL CONTRIBUTION
Neuroprotective effect of cocoa flavonids on in vitro oxidative stress
Emma Ramiro-Puig Gemma Casadess Hyoung-gon Lee Xiongwei ZhuAndrew McShea George Perry Francisco J. Prez-Cano Mark A. Smith Margarida Castell
j Abstract Background Cocoa is a rich source of avonoids that, among other functions, can act as antioxidants. In living systems, the production of reactive oxygen species (ROS) activate an array of intracellular cascades, including mitogen-activated protein kinases (MAPK), that are closely associated with cell death or survival pathways. Aim of the study To ascertain the role of a cocoa extract and its main avonoid, (-)-epicatechin, in an in vitro model of oxidative stress induced in a neuronal cell line. Methods We analyzed ROS production by uorometry (dichlorouorescein assay), and activation of MAPK pathways including extracellular signal-regulated kinases 1/2 (ERK 1/2), c-Jun N-terminal kinase (JNK), and p-38, by Western blot analysis. Results Cells incubated with cocoa extract or (-)-epicatechin, reduced ROS production in a dose-dependent manner, reaching 35% inhibition. pJNK and p38, involved in apoptosis, were down-modulated by cocoa extract and (-)-epicatechin
with p38 inhibition reaching up to 70%. Conclusions Our results show that cocoa extract and (-)-epicate-chin may exert a neuroprotective action by reducing ROS production and modulating MAPK activation.
j Key words cocoa
c-JNK MAPK neuronal
damage oxidative stress
Received: 16 September 2008 Accepted: 17 November 2008 Published online: 29 November 2008
E. Ramiro-Puig F.J. Prez-CanoM. Castell (&)
Dept. de FisiologiaFacultat de Farmcia (Universitat de Barcelona)
Av. Joan XXIII s/nBarcelona 08028, SpainE-Mail: [email protected]
G. CasadessDept. of NeuroscienceCase Western Reserve University Cleveland (OH), USA
H. Lee X. Zhu G. Perry M.A. Smith Dept. of PathologyCase Western Reserve University Cleveland (OH), USA
A. McSheaResearch and Development Theo ChocolateSeattle (WA), USA
G. PerryCollege of SciencesUniversity of Texas at San Antonio San Antonio (TX), USA
j Abbreviations AD: Alzheimers
disease, ANOVA: Analysis of variance, Ab: Amyloid-b protein, DA: diacetate, DCF: 2,7-dichlorouorescein, DMSO: Dimethyl sulfoxide, ERK 1/2: Extracellular signal-regulated kinases 1/2, GSTPi: Glutathione-S-transferase enzyme-class Pi, H2DCF: reduced DCF, JNK: c-Jun N-terminal kinase, LDH: Lactate dehydrogenase, MAPK: Mitogen-activated protein kinases, ORAC: Oxygen radical absorbance capacity, PMSF: Phenylmethylsulfonyl uo-ride, ROS: Reactive oxygen species, RT: Room temperature, SOD: Superoxide dismutase, TBS: Tris buffered saline solution, TBST: TBS with 0.1% Tween 20
Introduction
Aerobic organisms...