Skip to main content

Advertisement

SpringerLink
Log in

Neuroprotective effect of cocoa flavonids on in vitro oxidative stress

  • ORIGINAL CONTRIBUTION
  • Published:
European Journal of Nutrition Aims and scope Submit manuscript

An ERRATUM to this article was published on 10 December 2008

Abstract

Background

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.

Methods

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.

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 (-)-epicatechin may exert a neuroprotective action by reducing ROS production and modulating MAPK activation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Abbreviations

AD:

Alzheimer’s disease

ANOVA:

Analysis of variance

Aβ:

Amyloid-β protein

DA:

diacetate

DCF:

2′,7′-dichlorofluorescein

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 fluoride

ROS:

Reactive oxygen species

RT:

Room temperature

SOD:

Superoxide dismutase

TBS:

Tris buffered saline solution

TBST:

TBS with 0.1% Tween 20

References

  1. Andrés-Lacueva C, Lamuela-Raventós RM, Jáuregui O (2000) An LC method for the analysis of cocoa phenolics. LC-GC Eur 12:902–905

    Google Scholar 

  2. Canas N, Valero T, Villarroya M, Montell E, Verges J, Garcia AG, Lopez MG (2007) Chondrotitin sulfate protects SH-SY5Y cells from oxidative stress. J Pharmacol Exp Ther 323:946–953

    Article  CAS  Google Scholar 

  3. Casadesús G, Ramiro-Puig E, Webber KM, Atwood CS, Castell Escuer M, Bowen RL, Perry G, Smith MA (2006) Targeting gonadotropins: an alternative option for Alzheimer disease treatment. J Biomed Biotechnol 39508:1–8

    Article  CAS  Google Scholar 

  4. Cho ES, Lee KW, Lee HJ (2008) Cocoa procyanidins protect PC12 cells from hydrogen-peroxide-induced apoptosis by inhibiting activation of p38 MAPK and JNK. Mutat Res 640:123–130

    CAS  Google Scholar 

  5. Commenges D, Scotet V, Renaud S, Jacqmin-Gadda H, Barberger-Gateau P, Dartigues J-F (2000) Intake of flavonoids and risk of dementia. Eur J Epidemiol 16:357–363

    Article  CAS  Google Scholar 

  6. Cooper KA, Campos-Giménez E, Jiménez Alvarez D, Nagy K, Donovan JL, Williamson G (2007) Rapid reversed phase ultra-performance liquid chromatography analysis of the major cocoa polyphenols and inter-relationships of their concentrations in chocolate. J Agric Food Chem 55:2841–2847

    Article  CAS  Google Scholar 

  7. Cooper KA, Donovan JL, Waterhouse AL, Williamson G (2008) Cocoa and health: a decade of research. Br J Nutr 99:1–11

    Article  CAS  Google Scholar 

  8. Crews WD Jr, Harrison DW, Wright JW (2008) A double-blind, placebo-controlled, randomized trial of the effects of dark chocolate and cocoa on variables associated with neuropsychological functioning and cardiovascular health: clinical findings from a sample of healthy, cognitively intact older adults. Am J Clin Nutr 87:872–880

    CAS  Google Scholar 

  9. Dai Q, Borenstein AR, Wu Y, Jackson JC, Larson EB (2006) Fruit and vegetable juices and Alzheimer’s disease: the Kame project. Am J Med 119:751–759

    Article  CAS  Google Scholar 

  10. Datla KP, Zbarsky V, Rai D, Parkar S, Osakabe N, Aruoma OI, Dexter DT (2007) Short-term supplementation with plant extracts rich in flavonoids protect nigrostriatal dopaminergic neurons in a rat model of Parkinson’s disease. J Am Coll Nutr 26:341–349

    CAS  Google Scholar 

  11. de Boer VC, Dihal AA, van der Woude H, Arts IC, Wolffram S, Alink GM, Rietjens IM, Keijer J, Hollman PC (2005) Tissue distribution of quercetin in rats and pigs. J Nutr 135:1718–1725

    Google Scholar 

  12. Erlejman AG, Fraga CG, Oteiza PI (2006) Procyanidins protect caco-2 cells from bile acid- and oxidant-induced damage. Free Radic Biol Med 41:1247–1256

    Article  CAS  Google Scholar 

  13. Granado-Serrano AB, Martin MA, Bravo L, Goya L, Ramos S (2006) Quercetin induces apoptosis via caspase activation, regulation of bcl-2, and inhibition of PI-3-kinase/Akt and ERK pathways in a human hepatoma cell line (HepG2). J Nutr 136:2715–2721

    CAS  Google Scholar 

  14. Granado-Serrano AB, Martin MA, Izquierdo-Pulido M, Goya L, Bravo L, Ramos S (2007) Molecular mechanisms of (-)-epicatechin and chlorogenic acid on the regulation of the apoptotic and survival/proliferation pathways in a human hepatoma cell line. J Agric Food Chem 55:2020–2027

    Article  CAS  Google Scholar 

  15. Heo HJ, Lee CY (2005) Epicatechin and catechin in cocoa inhibit amyloid beta protein induced apoptosis. J Agric Food Chem 53:1445–1448

    Article  CAS  Google Scholar 

  16. Kamata H, Hirata H (1999) Redox regulation of cellular signalling. Cell Signal 11:1–14

    Article  CAS  Google Scholar 

  17. Lecumberri E, Goya L, Mateos R, Alía M, Ramos S, Izquierdo-Pulido M, Bravo L (2007) A diet rich in dietary fiber from cocoa improves lipid profile and reduces malondialdehyde in hypercholesterolemic rats. Nutrition 23:332–341

    Article  CAS  Google Scholar 

  18. McEligot AJ, Yang S, Meyskens FL Jr (2005) Redox regulation by intrinsic species and extrinsic nutrients in normal and cancer cells. Annu Rev Nutr 25:261–295

    Article  CAS  Google Scholar 

  19. McShea A, Ramiro-Puig E, Munro SB, Casadesus G, Castell M, Smith MA (2008) Clinical benefit and preservation of flavonols in dark chocolate manufacturing. Nutr Rev 66:630–641

    Article  Google Scholar 

  20. Miller KB, Stuart DA, Smith NL, Lee CY, McHale NL, Flanagan JA, Ou B, Hurst WJ (2006) Antioxidant activity and polyphenol and procyanidin contents of selected commercially available cocoa-containing and chocolate products in the United States. J Agric Food Chem 54:4062–4068

    Article  CAS  Google Scholar 

  21. Natsume M, Osakabe N, Yasuda A, Baba S, Tokunaga T, Kondo K, Osawa T, Terao J (2004) In vitro antioxidative activity of (-)-epicatechin glucuronide metabolites present in human and rat plasma. Free Radic Res 38:1341–1348

    Article  CAS  Google Scholar 

  22. Nijveldt RJ, van Nood E, van Hoorn DE, Boelens PG, Van Norren K, Van Leeuwen PAM (2001) Flavonoids: a review of probable mechanisms of action and potential applications. Am J Clin Nutr 74:418–425

    CAS  Google Scholar 

  23. Pensalfini A, Cecchi C, Zampagni M, Becatti M, Favilli F, Poli P, Catarzi S, Bagnoli S, Nacmias B, Sorbi S, Liguri G (2008) Protective effects of new derivatives against amyloid-induced oxidative stress. Free Radic Biol Med 44:1624–1636

    Article  CAS  Google Scholar 

  24. Pereda J, Sabater L, Cassinello N, Gómez-Cambronero L, Closa D, Folch-Puy E, Aparisi L, Calvete J, Cerda M, Lledo S, Vina J, Sastre J (2004) Effect of simultaneous inhibition of TNF-alpha production and xanthine oxidase in experimental acute pancreatitis: the role of mitogen activated protein kinases. Ann Surg 240:108–116

    Article  Google Scholar 

  25. Porter LJ, Ma Z, Chan BG (1991) Cacao procyanidins: major flavanoids and identification of some minor metabolites. Phytochemistry 30:1657–1663

    Article  CAS  Google Scholar 

  26. Pura Naik J (2001) Improved high-performance liquid chromatography method to determine theobromine and caffeine in cocoa and cocoa products. J Agric Food Chem 49:3579–3583

    Article  CAS  Google Scholar 

  27. Ramiro E, Franch A, Castellote C, Andrés-Lacueva C, Izquierdo-Pulido M, Castell M (2005) Effect of theobroma cacao flavonoids on immune activation of a lymphoid cell line. Br J Nutr 93:859–866

    Article  CAS  Google Scholar 

  28. Ramiro E, Franch A, Castellote C, Pérez-Cano FJ, Permanyer J, Izquierdo-Pulido M, Castell M (2005) Flavonoids from Theobroma cacao down-regulate inflammatory mediators. J Agric Food Chem 53:8506–8511

    Article  CAS  Google Scholar 

  29. Ramiro-Puig E, Urpi-Sarda M, Pérez-Cano FJ, Franch A, Castellote C, Andres-Lacueva C, Izquierdo-Pulido M, Castell M (2007) Cocoa-enriched diet enhances antioxidant enzyme activity and modulates lymphocyte composition in thymus from young rats. J Agric Food Chem 55:6431–6438

    Article  CAS  Google Scholar 

  30. Ramos S, Alia M, Bravo L, Goya L (2005) Comparative effects of food-derived polyphenols on the viability and apoptosis of a human hepatoma cell line (HepG2). J Agric Food Chem 53:1271–1280

    Article  CAS  Google Scholar 

  31. Raza H, John A (2005) Green tea polyphenol epigallocatechin-3-gallate differentially modulates oxidative stress in PC12 cell compartments. Toxicol Appl Pharmacol 207:212–220

    CAS  Google Scholar 

  32. Ruffels J, Griffin M, Dickenson JM (2004) Activation of ERK1/2, JNK and PKB by hydrogen peroxide in human SH-SY5Y neuroblastoma cells: role of ERK1/2 in H2O2-induced cell death. Eur J Pharmacol 483:163–173

    Article  CAS  Google Scholar 

  33. Sánchez-Rabaneda F, Jáuregui O, Casals I, Andrés-Lacueva C, Izquierdo-Pulido M, Lamuela-Raventos RM (2003) Liquid chromatographic/electrospray ionization tandem mass spectrometric study of the phenolic composition of cocoa (Theobroma cacao). J Mass Spectrom 38:35–42

    Article  CAS  Google Scholar 

  34. Schauss AG, Wu X, Prior RL, Ou B, Huang D, Owens J, Agarwal A, Jensen GS, Hart AN, Shanbrom E (2006) Antioxidant capacity and other bioactivities of the freeze-dried Amazonian palm berry, Euterpe oleraceae mart. (acai). J Agric Food Chem 54:8604–8610

    Article  CAS  Google Scholar 

  35. Smith MA, Hirai K, Hsiao K, Pappolla MA, Harris PL, Siedlak SL, Tabaton M, Perry G (1998) Amyloid-beta deposition in Alzheimer transgenic mice is associated with oxidative stress. J Neurochem 70:2212–2215

    CAS  Google Scholar 

  36. Spencer JP, Abd-el-Mohsen MM, Rice-Evans C (2004) Cellular uptake and metabolism of flavonoids and their metabolites: implications for their bioactivity. Arch Biochem Biophys 423:148–161

    Article  CAS  Google Scholar 

  37. Tomas-Barberan FA, Cienfuegos-Jovellanos E, Marín A, Muguerza B, Gil-Izquierdo A, Cerda B, Zafrilla P, Morillas J, Mulero J, Ibarra A, Pasamar MA, Ramón D, Espín JC (2007) A new process to develop a cocoa powder with higher flavonoid monomer content and enhanced bioavailability in healthy humans. J Agric Food Chem 55:3926–3935

    Article  CAS  Google Scholar 

  38. Uberti D, Piccioni L, Colzi A, Bravi D, Canonico PL, Memo M (2002) Pergolide protects SH-SY5Y cells against neurodegeneration induced by H2O2. Eur J Pharmacol 434:17–20

    Article  CAS  Google Scholar 

  39. Waetzig V, Herdegen T (2005) Context-specific inhibition of JNKs: overcoming the dilemma of protection and damage. Trends Pharmacol Sci 26:455–461

    CAS  Google Scholar 

  40. Wang H, Joseph JA (1999) Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader. Free Radic Biol Med 27:612–616

    Article  CAS  Google Scholar 

  41. Wollgast J, Anklam E (2000) Review on polyphenols in Theobroma cacao: changes in composition during the manufacture of chocolate and methodology for identification and quantification. Food Res Int 33:423–447

    Article  CAS  Google Scholar 

  42. Yilmaz Y, Toledo RT (2004) Major flavonoids in grape seeds and skins: antioxidant capacity of catechin, epicatechin, and gallic acid. J Agric Food Chem 52:255–260

    Article  CAS  Google Scholar 

  43. Youdim KA, Dobbie MS, Kuhnle G, Proteggente AR, Abbott NJ, Rice-Evans C (2003) Interaction between flavonoids and the blood-brain barrier: in vitro studies. J Neurochem 85:180–192

    Article  CAS  Google Scholar 

Download references

Acknowledgments

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

Author information

Authors and Affiliations

  1. Dept. de Fisiologia, Facultat de Farmàcia (Universitat de Barcelona), Av. Joan XXIII s/n, 08028, Barcelona, Spain

    Emma Ramiro-Puig, Francisco J. Pérez-Cano & Margarida Castell

  2. Dept. of Neuroscience, Case Western Reserve University, Cleveland, OH, USA

    Gemma Casadesús

  3. Dept. of Pathology, Case Western Reserve University, Cleveland, OH, USA

    Hyoung-gon Lee, Xiongwei Zhu, George Perry & Mark A. Smith

  4. Research and Development, Theo Chocolate, Seattle, WA, USA

    Andrew McShea

  5. College of Sciences, University of Texas at San Antonio, San Antonio, TX, USA

    George Perry

Authors
  1. Emma Ramiro-Puig
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gemma Casadesús
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hyoung-gon Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xiongwei Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Andrew McShea
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. George Perry
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Francisco J. Pérez-Cano
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Mark A. Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Margarida Castell
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Margarida Castell.

Additional information

An erratum to this article can be found under doi:10.1007/s00394-008-0766-z.

An erratum to this article can be found at http://dx.doi.org/10.1007/s00394-008-0766-z

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ramiro-Puig, E., Casadesús, G., Lee, Hg. et al. Neuroprotective effect of cocoa flavonids on in vitro oxidative stress. Eur J Nutr 48, 54–61 (2009). https://doi.org/10.1007/s00394-008-0761-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00394-008-0761-4

Keywords

Search

Navigation