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DNA methylation regulates gene expression and can be modified by different bioactive compounds in foods, such as polyphenols. Cocoa is a rich source of polyphenols, but its role in DNA methylation is still unknown. The objective was to assess the effect of cocoa consumption on DNA methylation and to determine whether the enzymes involved in the DNA methylation process participate in the mechanisms by which cocoa exerts these effects in humans. The global DNA methylation levels in the peripheral blood were evaluated in 214 volunteers who were pre-hypertensive, stage-1 hypertensive or hypercholesterolemic. The volunteers were divided into two groups: 110 subjects who consumed cocoa (6 g/d) for two weeks and 104 control subjects. In addition, the peripheral blood mononuclear cells (PBMCs) from six subjects were treated with a cocoa extract to analyze the mRNA levels of the DNA methyltransferases ( DNMTs ), methylenetetrahydrofolate reductase ( MTHFR ), and methionine synthase reductase ( MTRR ) genes. Cocoa consumption significantly reduced the DNA methylation levels (2.991±0.366 vs. 3.909±0.380, p<0.001). Additionally, we found an association between the cocoa effects on DNA methylation and three polymorphisms located in the  MTHFR ,  MTRR , and  DNMT3B  genes. Furthermore, in PBMCs, the cocoa extract significantly lowered the mRNA levels of the  DNMTs ,  MTHFR , and  MTRR . Our study demonstrates for the first time that the consumption of cocoa decreases the global DNA methylation of peripheral leukocytes in humans with cardiovascular risk factors.  In vitro  experiments with PBMCs suggest that cocoa may exert this effect partially via the down-regulation of  DNMTs ,  MTHFR  and  MTRR , which are key genes involved in this epigenetic process.   Trial Registration    Clinicaltrials.gov   NCT00511420  and  NCT00502047

plos one

Cocoa Consumption Alters the Global DNA Methylation of Peripheral Leukocytes in Humans with Cardiovascular Disease Risk Factors: A Randomized Controlled Trial