Top

Published in:

Open Access 01-08-2019 | Original Contribution

Gene expression changes by high-polyphenols cocoa powder intake: a randomized crossover clinical study

Authors: P. K. Barrera-Reyes, N. Hernández-Ramírez, J. Cortés, L. Poquet, K. Redeuil, C. Rangel-Escareño, M. Kussmann, I. Silva-Zolezzi, M. E. Tejero

Published in: European Journal of Nutrition | Issue 5/2019

Abstract

Purpose

To assess the effect of the intake of a single dose of high-polyphenols cocoa on gene expression in peripheral mononuclear cells (PBMCs), and analyze conjugated (−)-epicatechin metabolites in plasma, which may be related with an antioxidant response in healthy human.

Methods

A randomized, controlled, double-blind, cross-over, clinical trial in healthy young adults who consumed a single dose of high-polyphenols cocoa powder and maltodextrins as control, with a one-week washout period. Analysis of circulating metabolites, plasma antioxidant capacity and gene expression changes in PBMCs were performed under fasting conditions and 2-h after treatment using microarray in a subsample. Pathway analysis was conducted using Ingenuity Pathway Analysis (IPA).

Results

Twenty healthy participants (9 F) were included in the study. A significant increase in circulating (−)-epicatechin metabolites was found after cocoa intake in all participants without related changes in antioxidant capacity of plasma. The metabolites profile slightly varied across subjects. Treatments triggered different transcriptional changes in PBMC. A group of 98 genes showed changes in expression after cocoa treatment, while only 18 were modified by control. Differentially expressed genes included inflammatory cytokines and other molecules involved in redox balance. Gene and network analysis after cocoa intake converged in functions annotated as decreased production of reactive oxygen species (p = 9.58E−04), decreased leukocyte activation (p = 4E−03) and calcium mobilization (p = 2.51E–05).

Conclusions

No association was found between conjugated metabolites in plasma and antioxidant capacity. Changes in PBMCs gene expression suggest anti-inflammatory effects.

Available only for authorised users

Corti R et al (2009) Cocoa and cardiovascular health. Circulation. 119(10):1433–1441PubMedCrossRef

Buitrago-Lopez A et al (2011) Chocolate consumption and cardiometabolic disorders: systematic review and meta-analysis. BMJ 343:d4488PubMedPubMedCentralCrossRef

Andujar I et al (2012) Cocoa polyphenols and their potential benefits for human health. Oxidative Medicine and Cellular Longevity

Blumberg JB et al (2014) The science of cocoa flavanols: bioavailability, emerging evidence, and proposed mechanisms. Adv Nutr 5(5):547–549PubMedPubMedCentralCrossRef

Murphy KJ et al (2003) Dietary flavanols and procyanidin oligomers from cocoa (Theobroma cacao) inhibit platelet function. Am J Clin Nutr 77(6):1466–1473PubMedCrossRef

Scalbert A, Johnson IT, Saltmarsh M (2005) Polyphenols: antioxidants and beyond. Am J Clin Nutr 81(1 Suppl):215S–217SPubMedCrossRef

Ramiro-Puig E et al (2007) Cocoa-enriched diet enhances antioxidant enzyme activity and modulates lymphocyte composition in thymus from young rats. J Agric Food Chem 55(16):6431–6438PubMedCrossRef

Steffen Y, Schewe T, Sies H (2007) (-)-Epicatechin elevates nitric oxide in endothelial cells via inhibition of NADPH oxidase. Biochem Biophys Res Commun 359(3):828–833.PubMedCrossRef

Cooper KA et al (2008) Cocoa and health: a decade of research. Br J Nutr 99(1):1–11PubMedCrossRef

10.

Duenas M et al (2010) Antioxidant evaluation of O-methylated metabolites of catechin, epicatechin and quercetin. J Pharm Biomed Anal 51(2):443–449PubMedCrossRef

11.

Actis-Goretta L et al (2012) Elucidation of (−)-epicatechin metabolites after ingestion of chocolate by healthy humans. Free Radic Biol Med 53(4):787–795PubMedCrossRef

12.

Ramiro E et al (2005) Effect of Theobroma cacao flavonoids on immune activation of a lymphoid cell line. Br J Nutr 93(6):859–866PubMedCrossRef

13.

Erlejman AG et al (2008) TNFalpha-induced NF-kappaB activation and cell oxidant production are modulated by hexameric procyanidins in Caco-2 cells. Arch Biochem Biophys 476(2):186–195PubMedCrossRef

14.

Verstraeten SV et al (2008) (−)-Epicatechin and related procyanidins modulate intracellular calcium and prevent oxidation in Jurkat T cells. Free Radic Res 42(10):864–872PubMedCrossRef

15.

Granado-Serrano AB et al (2010) Epicatechin induces NF-kappaB, activator protein-1 (AP-1) and nuclear transcription factor erythroid 2p45-related factor-2 (Nrf2) via phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) and extracellular regulated kinase (ERK) signalling in HepG2 cells. Br J Nutr 103(2):168–79PubMedCrossRef

16.

Mao T et al (2000) Cocoa procyanidins and human cytokine transcription and secretion. J Nutr 130(8S Suppl):2093S–2099SPubMedCrossRef

17.

Mao TK et al (2002) Modulation of TNF-α secretion in peripheral blood mononuclear cells by cocoa flavanols and procyanidins. Dev Immunol 9(3):135–141PubMedPubMedCentralCrossRef

18.

Ramiro-Puig E et al (2007) Spleen lymphocyte function modulated by a cocoa-enriched diet. Clin Exp Immunol 149(3):535–542PubMedPubMedCentralCrossRef

19.

Zempo H et al (2016) Cacao polyphenols ameliorate autoimmune myocarditis in mice. Hypertens Res 39(4):203–209PubMedCrossRef

20.

Kroon PA et al (2004) How should we assess the effects of exposure to dietary polyphenols in vitro? Am J Clin Nutr 80(1):15–21PubMedCrossRef

21.

Ottaviani JI et al (2016) The metabolome of [2-(14)C](−)-epicatechin in humans: implications for the assessment of efficacy, safety, and mechanisms of action of polyphenolic bioactives. Sci Rep 6:29034PubMedPubMedCentralCrossRef

22.

Goya L et al (2016) Effect of cocoa and its flavonoids on biomarkers of inflammation: studies of cell culture. Anim Hum Nutr 8(4):212

23.

Olsen KS, Skeie G, Lund E (2015) Whole-blood gene expression profiles in large-scale epidemiological studies: what do they tell? Curr Nutr Rep 4(4):377–386PubMedPubMedCentralCrossRef

24.

Monagas M et al (2009) Effect of cocoa powder on the modulation of inflammatory biomarkers in patients at high risk of cardiovascular disease. Am J Clin Nutr 90(5):1144–1150PubMedCrossRef

25.

Vazquez-Agell M et al (2013) Cocoa consumption reduces NF-kappaB activation in peripheral blood mononuclear cells in humans. Nutr Metab Cardiovasc Dis 23(3):257–263PubMedCrossRef

26.

Craig CL et al (2003) International physical activity questionnaire: 12-country reliability and validity. Med Sci Sports Exerc 35(8):1381–1395PubMedCrossRef

27.

Morey JS, Ryan JC, Van Dolah FM (2006) Microarray validation: factors influencing correlation between oligonucleotide microarrays and real-time PCR. Biol Proced Online 8:175–193PubMedPubMedCentralCrossRef

28.

Irizarry RA et al (2003) Summaries of affymetrix GeneChip probe level data. Nucleic Acids Res 31(4):e15PubMedPubMedCentralCrossRef

29.

Bolstad BM et al (2003) A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics 19(2):185–193PubMedCrossRef

30.

Smyth GK (2004) Linear models and empirical bayes methods for assessing differential expression in microarray experiments. Stat Appl Genet Mol Biol 3:Article3CrossRefPubMed

31.

Ritchie ME et al (2015) limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res 43(7):e47PubMedPubMedCentralCrossRef

32.

Benjamini Y et al (2001) Controlling the false discovery rate in behavior genetics research. Behav Brain Res 125(1–2):279–284PubMedCrossRef

33.

Suzuki R, Shimodaira H (2006) Pvclust: an R package for assessing the uncertainty in hierarchical clustering. Bioinformatics 22(12):1540–1542PubMedCrossRef

34.

Kramer A et al (2014) Causal analysis approaches in ingenuity pathway analysis. Bioinf 30(4):523–530CrossRef

35.

Lakens D (2013) Calculating and reporting effect sizes to facilitate cumulative science: a practical primer for t-tests and ANOVAs. Front Psychol 4:863PubMedPubMedCentralCrossRef

36.

Faul F et al (2007) G*Power 3: a flexible statistical power analysis program for the social, behavioral, and biomedical sciences. Behav Res Methods 39(2):175–191PubMedCrossRef

37.

Vogiatzoglou A et al (2014) Assessment of the dietary intake of total flavan-3-ols, monomeric flavan-3-ols, proanthocyanidins and the aflavins in the European Union. Br J Nutr 111(8):1463–1473PubMedCrossRef

38.

Rodriguez-Mateos A et al. (2015) Interactions between cocoa flavanols and inorganic nitrate: additive effects on endothelial function at achievable dietary amounts. Free Radic Biol Med 80:121–128PubMedCrossRef

39.

Vogiatzoglou A et al (2015) Associations between flavan-3-ol intake and CVD risk in the Norfolk cohort of the European Prospective Investigation into Cancer (EPIC-Norfolk). Free Radical Biol Med 84:1–10CrossRef

40.

Rodriguez-Mateos A et al (2014) Uptake and metabolism of (−)-epicatechin in endothelial cells. Arch Biochem Biophys 559:17–23PubMedCrossRef

41.

Franconi F, Campesi I (2014) Pharmacogenomics, pharmacokinetics and pharmacodynamics: interaction with biological differences between men and women. Br J Pharmacol 171(3):580–594PubMedPubMedCentralCrossRef

42.

Manach C et al (2017) Addressing the inter-individual variation in response to consumption of plant food bioactives: Towards a better understanding of their role in healthy aging and cardiometabolic risk reduction. Mol Nutr Food Res 61(6):1600557CrossRef

43.

Waxman DJ, Holloway MG (2009) Sex differences in the expression of hepatic drug metabolizing enzymes. Mol Pharmacol 76(2):215–228PubMedPubMedCentralCrossRef

44.

Ottaviani JI et al (2012) Structurally related (−)-epicatechin metabolites in humans: assessment using de novo chemically synthesized authentic standards. Free Radical Biol Med 52(8):1403–1412CrossRef

45.

Lotito SB, B. Frei (2006) Consumption of flavonoid-rich foods and increased plasma antioxidant capacity in humans: cause, consequence, or epiphenomenon? Free Radic Biol Med 41(12):1727–1746PubMedCrossRef

46.

Wiswedel I et al (2004) Flavanol-rich cocoa drink lowers plasma F(2)-isoprostane concentrations in humans. Free Radic Biol Med 37(3):411–421PubMedCrossRef

47.

Spadafranca A et al (2010) Effect of dark chocolate on plasma epicatechin levels, DNA resistance to oxidative stress and total antioxidant activity in healthy subjects. Br J Nutr 103(7):1008–1014PubMedCrossRef

48.

Fraga CG, Oteiza PI (2011) Dietary flavonoids: role of (−)-epicatechin and related procyanidins in cell signaling. Free Radic Biol Med 51(4):813–823PubMedCrossRef

49.

Chen L et al (2017) Intracellular signaling pathways of inflammation modulated by dietary flavonoids: The most recent evidence. Crit Rev Food Sci Nutr 6:1–17

50.

Tome-Carneiro J et al (2013) One-year supplementation with a grape extract containing resveratrol modulates inflammatory-related microRNAs and cytokines expression in peripheral blood mononuclear cells of type 2 diabetes and hypertensive patients with coronary artery disease. Pharmacol Res 72:69–82PubMedCrossRef

51.

Milenkovic D et al (2014) Dietary flavanols modulate the transcription of genes associated with cardiovascular pathology without changes in their DNA methylation state. PLoS One 9(4):e95527PubMedPubMedCentralCrossRef

52.

Cialdella-Kam L et al (2016) A mixed flavonoid-fish oil supplement induces immune-enhancing and anti-inflammatory transcriptomic changes in adult obese and overweight women-a randomized controlled trial. Nutrients 8(5)

53.

Martin-Pelaez S et al (2017) Effect of olive oil phenolic compounds on the expression of blood pressure-related genes in healthy individuals. Eur J Nutr 56(2):663–670PubMedCrossRef

54.

Keen CL et al (2005) Cocoa antioxidants and cardiovascular health. Am J Clin Nutr 81(1 Suppl):298S–303SPubMedCrossRef

55.

Whitney AR et al (2003) Individuality and variation in gene expression patterns in human blood. Proc Natl Acad Sci USA 100(4):1896–901PubMedCrossRefPubMedCentral

56.

Milenkovic D et al (2014) Dietary flavanols modulate the transcription of genes associated with cardiovascular pathology without changes in their DNA methylation state. Plos One 9(4)

57.

Kim CH et al (2008) Epicatechin protects auditory cells against cisplatin-induced death. Apoptosis 13(9):1184–1194PubMedCrossRef

58.

Shin HA et al. (2014) Radioprotective effect of epicatechin in cultured human fibroblasts and zebrafish. J Radiat Res 55(1):32–40PubMedCrossRef

59.

Afman L, Milenkovic D, Roche HM (2014) Nutritional aspects of metabolic inflammation in relation to health–insights from transcriptomic biomarkers in PBMC of fatty acids and polyphenols. Mol Nutr Food Res 58(8):1708–20PubMedCrossRef

60.

Lipton RB, Bigal ME, Stewart WF (2005) Clinical trials of acute treatments for migraine including multiple attack studies of pain, disability, and health-related quality of life. Neurology 65(12 Suppl 4):S50–S58PubMedCrossRef

Title: Gene expression changes by high-polyphenols cocoa powder intake: a randomized crossover clinical study
Authors: P. K. Barrera-Reyes
N. Hernández-Ramírez
J. Cortés
L. Poquet
K. Redeuil
C. Rangel-Escareño
M. Kussmann
I. Silva-Zolezzi
M. E. Tejero
Publication date: 01-08-2019
Publisher: Springer Berlin Heidelberg
Published in: European Journal of Nutrition / Issue 5/2019
Print ISSN: 1436-6207
Electronic ISSN: 1436-6215
DOI: https://doi.org/10.1007/s00394-018-1736-8

ACC 2024 Congress Coverage

Heart Failure Video

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Gene expression changes by high-polyphenols cocoa powder intake: a randomized crossover clinical study

Abstract

Purpose

Methods

Results

Conclusions

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 5/2019

Food groups and risk of age-related macular degeneration: a systematic review with meta-analysis

An open-label, randomized, placebo-controlled study on the effectiveness of a novel probiotics administration protocol (ProbiotiCKD) in patients with mild renal insufficiency (stage 3a of CKD)

Pharmacokinetics of vitamin E, γ-oryzanol, and ferulic acid in healthy humans after the ingestion of a rice bran-enriched porridge prepared with water or with milk

Selenium and sulphur derivatives of hydroxytyrosol: inhibition of lipid peroxidation in liver microsomes of vitamin E-deficient rats

Еffects of fortified milk on cognitive abilities in school-aged children: results from a randomized-controlled trial

Effects of vitamin D supplementation on metabolic and endocrine parameters in PCOS: a randomized-controlled trial

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page