European Journal of Nutrition

, Volume 48, Issue 1, pp 54–61 | Cite as

Neuroprotective effect of cocoa flavonids on in vitro oxidative stress

  • Emma Ramiro-Puig
  • Gemma Casadesús
  • Hyoung-gon Lee
  • Xiongwei Zhu
  • Andrew McShea
  • George Perry
  • Francisco J. Pérez-Cano
  • Mark A. Smith
  • Margarida Castell



Cocoa is a rich source of flavonoids 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 flavonoid, (-)-epicatechin, in an in vitro model of oxidative stress induced in a neuronal cell line.


We analyzed ROS production by fluorometry (dichlorofluorescein 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.


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%.


Our results show that cocoa extract and (-)-epicatechin may exert a neuroprotective action by reducing ROS production and modulating MAPK activation.


cocoa c-JNK MAPK neuronal damage oxidative stress 



Alzheimer’s disease


Analysis of variance

Amyloid-β protein






Dimethyl sulfoxide

ERK 1/2

Extracellular signal-regulated kinases 1/2


Glutathione-S-transferase enzyme-class Pi


reduced DCF


c-Jun N-terminal kinase


Lactate dehydrogenase


Mitogen-activated protein kinases


Oxygen radical absorbance capacity


Phenylmethylsulfonyl fluoride


Reactive oxygen species


Room temperature


Superoxide dismutase


Tris buffered saline solution


TBS with 0.1% Tween 20



E.R. is the recipient of fellowships from the Generalitat de Catalunya (2003FI 00578 and 2005BE 00709).


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Copyright information

© Spinger 2008

Authors and Affiliations

  • Emma Ramiro-Puig
    • 1
  • Gemma Casadesús
    • 2
  • Hyoung-gon Lee
    • 3
  • Xiongwei Zhu
    • 3
  • Andrew McShea
    • 4
  • George Perry
    • 3
    • 5
  • Francisco J. Pérez-Cano
    • 1
  • Mark A. Smith
    • 3
  • Margarida Castell
    • 1
  1. 1.Dept. de FisiologiaFacultat de Farmàcia (Universitat de Barcelona)BarcelonaSpain
  2. 2.Dept. of NeuroscienceCase Western Reserve UniversityClevelandUSA
  3. 3.Dept. of PathologyCase Western Reserve UniversityClevelandUSA
  4. 4.Research and DevelopmentTheo ChocolateSeattleUSA
  5. 5.College of SciencesUniversity of Texas at San AntonioSan AntonioUSA

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