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FOOD COMPONENTS AFFECTING THE EPIGENOME:

“ERGOGENETIC” AIDS FOR PERFORMANCE

Andrea G. Izquierdo, Manuel Portela, Paula M. Lorenzo, Federico

Mallo, Ana B. Crujeiras

PII: S2213-4344(20)30056-6
DOI: https://doi.org/10.1016/j.phanu.2020.100231
Reference: PHANU 100231

To appear in: PharmaNutrition

Received Date: 18 May 2020

Revised Date: 17 September 2020
Accepted Date: 29 September 2020

Please cite this article as: Izquierdo AG, Portela M, Lorenzo PM, Mallo F, Crujeiras AB, FOOD
COMPONENTS AFFECTING THE EPIGENOME: “ERGOGENETIC” AIDS FOR
PERFORMANCE, PharmaNutrition (2020), doi: https://doi.org/10.1016/j.phanu.2020.100231

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© 2020 Published by Elsevier.

TITLE: FOOD COMPONENTS AFFECTING THE EPIGENOME:

"ERGOGENETIC" AIDS FOR PERFORMANCE

Andrea G Izquierdo1,2*, Manuel Portela 3*, Paula M Lorenzo1*, Federico Mallo3#, Ana B

Crujeiras1,2#

1Epigenomics in Endocrinology and Nutrition Group, Instituto de Investigacion Sanitaria

(IDIS), Complejo Hospitalario Universitario de Santiago (CHUS/SERGAS), Santiago de

Compostela, Spain.

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2CIBER Fisiopatologia de la Obesidad y Nutricion (CIBERobn), Madrid, Spain.

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3 Laboratory of Endocrinology (LabEndo). Biomedical Research Center (CINBIO) –

University of Vigo – e-36310 – Vigo - Spain.

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*These authors contributed equally to this work and should be considered co-first authors.
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#Both authors contributed equally to this work and should be considered co-main authors.
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CORRESPONDENCE: Dra. Ana B. Crujeiras. Laboratory of Epigenomics in

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Endocrinology and Nutrition. EpigenomicsUnit. Instituto de Investigación

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Sanitaria,Complejo Hospitalario Universitario de Santiago (CHUS). Travesía da

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Choupana Street s/n, 15706 Santiago de Compostela, La Coruña, Spain; E-mail:

anabelencrujeiras@hotmail.com. Tel: +34 981955069.

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KEY WORDS: exercise, dietary supplementation, athletes, DNA methylation,

nutriepigenetic, epigenetic diet.

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ABSTRACT

The interest in epigenetics has grown exponentially in recent years and this is because
epigenetic marks are present in most life processes. It is known that the epigenome is
determined by different environmental factors such as diet or exercise. For athletes,
improving performance during physical activity or reducing recovery times after training
are key priorities that are currently focusing on research in the world of sports. Nutritional
strategies are important tools for this purpose, since more and more dietary supplements
are used as ergogenic potentials. Therefore and given that nutrition modulates gene
expression through epigenetic modifications, recent studies have aimed to elucidate
possible epigenetic mechanisms responsible for the link between dietary supplements for

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athletes and improving performance during exercise. Thus, in this review we cite the latest

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most relevant studies carried out to date in relation to epigenetic marks associated with
sports supplementation and performance.

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INTRODUCTION

In the last decade, epigenetics has experienced a growing interest within the scientific
community since it participates in most of the organism's biological processes [1]. During
development, epigenetic marks participate in cell differentiation, which give the cells
different characteristics and functions despite containing the same genetic information
[2]. In general terms, epigenetics is defined as a branch of science that is responsible for
studying molecular bases through external agents modulating gene expression
(phenotype), without implying an alteration of the DNA sequence (genotype) [3].
Epigenetics refers to alternative modifications to the DNA chain that exerts a direct effect
on transcriptional levels, causing the activation or inactivation of genes [4]. Numerous

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cellular processes are influenced by epigenetic modifications, hence the importance of

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systematically understanding this regulation on cellular machinery [5] and the
pathogenesis of the disease [6–8]. Epigenetic marks are stable and can be transmitted to
the next generations [9], although they can also change in response to specific stimuli to

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the organism providing a dynamic response in a short period of time [10]. Thus, these
marks are characterized by being acquired, inheritable and reversible [11]. Among the
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main most relevant mechanisms in epigenetic regulation are DNA methylation, post-
translational histone modifications (PTM) and non-coding RNA (ncRNA), which
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includes microRNA (miRNA) and long non-coding RNA (lncRNA) [4,12]. Among these,
DNA methylation is the most abundant epigenetic mechanism in the body and the most
studied [13].
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It has been shown that both the environment and lifestyle play a very important role in
gene expression [14]. Among the main determinants of health, genetics only contributes
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30% to the existing variability, while the remaining 70% depends on environmental
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factors [15]. Recent research shows that exposure of the organism to certain external
agents, such as diet and physical exercise, can trigger alterations in the epigenome [16–
18]. This implies that nutrients and physical activity influence the expression of genes
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and therefore have a relevant role in the state of health and disease prevention in
individuals. In this context, the most widely accepted current hypothesis postulates
epigenetic marks as the molecular machinery responsible for these processes [19,20]. As
an example, it was hypothesized that epigenetic dysregulation could contribute to a rapid
increase in the prevalence of obesity and its complications due to the heritability of
acquired epigenetic marks [21]. Both global DNA methylation and the methylation levels

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of specific genes related to adipose tissue function and metabolism in visceral adipose
tissue were found to be related to the metabolic syndrome aetiology [22,23] and insulin
resistance [7]. Moreover, it was evidenced that the association between obesity and cancer
can be mediated by epigenetic mechanisms because a specific methylome was observed
for example in tumors from breast [24] or colon [25] depending on adiposity. These
epigenetic marks associated with the pathogenesis of obesity also link with the beneficial
effect of different therapeutic strategies to lose weight such as hypocaloric diets [26,27]
or bariatric surgery [8,28], even after physical activity in patients with obesity [29,30].
Nutritional compounds or dietary patterns such as Mediterranean diet or other bioactive
components are able to modulate epigenetic marks to induce healthy effects [4].
In a similar way, in the field of exercise training and physical performance the relationship

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of epigenetic modulations with acute and chronic effect of exercise training was also
evidenced [31,32]. Relevantly, in the last decades it is usual the consumption of different
nutritional compounds or ergogenic aids among athletes to improve their

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performance[33]. The effect of these nutritional and non-nutritional components on
muscle function during exercise performance could be mediated by epigenetic
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mechanisms.
In this work we review the latest work on epigenetic mechanisms associated with the
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effect of bioactive components of sports supplementation and dietary patterns on

improving performance in physical training.
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EFFECT OF DIET, NUTRIENTS AND BIOACTIVE COMPOUNDS ON
EXERCISE PERFORMANCE

Dietary habits are among the factors that influence sports performance [34]. Nutritional
interventions may further augment high-intensity training by enhancing energy
metabolism during exercise [35]. In this context, Mediterranean diet is postulated as one
of the healthiest eating patterns due to the high consumption of plant-based foods, olive
oil, a certain number of dairy products, as well as a style active life [34]. Recent studies
have showed a positive relation between a high performance and a high adherence to
Mediterranean diet [36]. Adequate dietary intake and the correct balance of
macronutrients and micronutrients, with appropriate timing to enhance performance and

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recovery, will enable athletes to train and perform optimally [37].

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Macronutrients
The primary component to optimize training and performance through nutrition is to

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ensure the athlete is consuming enough calories to offset energy expenditure produced by
training [38,39]. In this way, the caloric needs of elite athletes can approach 40-70
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kcals/kg/day [38], in addition to needing adequate amounts of carbohydrates, proteins and
fat [39]. Carbohydrate intake is used by athletes as fuel to enhance athletic performance
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by reducing fatigue [40], to support the immune system, and to improve the
bioavailability of other supplements (e.g., protein). When exercising at high intensities,
carbohydrates become the main energy substrate, and thus glycogen depletion is
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considered an important limiting factor of performance [41]. In particular and as it relates

to exercise performance, the need for optimal glycogen store before, during and after
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intense and high-volume bouts of training and competition is evident [38]. In this way,
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current reviews and research highlight the importance of carbohydrate intake for athletes
competing in various endurance activities or team sports [40,42,43].
Other relevant macronutrients in the success of sports performance are dietary proteins.
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There is a large amount of literature that suggests that higher amounts of protein are
needed when exercising at high intensity to optimize athletic performance. Although there
is some variability in the data on the necessary amounts of protein, these sources indicate
that people who undergo intense training should consume between 1.4-2.0 g of protein
per kilogram of body weight per day, above the recommended 0.8-1 g for the general
population [35,38]. In this way, it is guaranteed the synthesis of muscle proteins, the

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maintenance of muscle mass and recovery after exercise [39]. Moreover, whey protein
has had a great impact as a nutritional supplement for athletes, as it contains almost 50%
of essential amino acids and approximately 26% of branched chain amino acids. This
amino acid composition has a pattern similar to the amino acid composition of human
skeletal muscle, so it is absorbed more quickly than other protein sources [44]. Regardless
of the source of protein or type of food, research indicates that the intake of 6-20 g of
protein before and/or after resistance exercise might stimulates protein synthesis [38,45].
This amount of protein can easily achieved with a proper balance meal. On a daily basis
may range 1.2-2g/kg/day [46]. Regarding to fat, although there are contradictory results,
it has been observed that supplementation with fatty acids, specifically omega-3
polyunsaturated fatty acids and extra virgin olive oil, reduces exercise-induced

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inflammation [47].
Additionally, there are several nutritional compounds obtained from food or provided as
supplements that in specific conditions may improve performance in athletes. Those with

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some scientific evidence are reported in Table 1, despite about most of them there is not
consistent support to be used in common conditions or to be generally recommended to
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athletes as part of their normal nutritional plan.
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Micronutrients
Deficient of common vitamins and minerals would markedly reduce athletic performance
and even affect the health of the sportsmen/women. Intense exercise, increase the risk of
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some common mineral deficits, as a trend to anaemia because iron insufficient supply,
and iodine deficiency in athletes living in continental regions with not easy access to fish
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and seafood. As general population, 2/3 of athletes are in risk of moderate vitamin D3
deficiency, which also may greatly affect performance and recovery after efforts [39].
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However, there is no clear scientific evidence about the role of vitamins and minerals in
sports performance when additionally given as supplements. Research focuses on
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whether an athlete is deficient in vitamins or minerals, supplements or dietary

modifications to achieve optimal amounts of these nutrients can improve health and
performance. In particular, acute changes in sodium, potassium and magnesium during
moderate to high intensity exercise are important features to consider [38]. As happens
with macronutrients, the athlete´s demand of micronutrients are achieved with a healthy
balance diet.

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The best evidence that exists in relation to the role of micronutrients on performance is
in relation to iron. Iron is the metal needed to configure hemoglobin heme-group,
allowing oxygen transport. In addition, iron is an important co-factor to several enzymes
involved in muscle function and repair. The iron deficiency may limit exercise work
capacity, which would lead to a compromised adaptation to training and reduced
performance [48]. Iron supplementation has been used to help increase aerobic
performance through the restoration of hemoglobin concentrations and thus improve
oxygen carrying capacity [49], although there is controversy in this regard [50]. In relation
to iron intake, its absorption rate in gut is in the range of 5-25% 14 depending whether is
non-heme iron (Fe +++, ferric) from vegetables or heme iron (Fe ++, ferrous) coming
from animal sources. 90-95% of the absorbed iron is linked to animal protein15. A

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balanced diet supplies approximately 6 mg of iron per 1000 kcal16, enough to cover
athletes daily requirements (about 20 mg/day).

Cholecalciferols are the food precursors of vitamin D3. This molecule has an action

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mechanism similar to all other steroid hormones, regulating the transcription of the genes
having vitamin D3 responsive elements (DRE) in their promotor sequences. Usually
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vitamin D3 interacts with thyroid hormone and estrogens in regulating these genes.
Vitamin D3 has many relevant biological actions to keep physiological function and
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homeostasis in all tissues. Its action is critical to maintain respiratory chain in

mitochondria and energy metabolism. Vitamin D3 regulates the absorption and
metabolism of calcium and phosphorus and plays a key role in the maintenance of bone
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health. Some studies have reported that normal levels of vitamin D3 prevent injuries in
sports [51], accelerate rehabilitation [52] ameliorate neuromuscular function [53],
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promote hypertrophy of the type II muscle fibres, reduce inflammation and diminish the
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risk of stress fractures [54].

Ergogenic aids
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Caffeine
Caffeine is consumed by general population as part of occidental diets [55] that may
improve high-intensity boots of athletic performance for short times [35,56]. A large
number of studies have shown the effectiveness of caffeine supplementation on athletic
performance in both aerobic [57], anaerobic [58, 59] or mixed [60] activities. Among the
highlighted effects are the reduction of pain and exertion perception, improvement of
muscle relaxation time by optimizing calcium mobilization and improvement of muscle

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function [35]. There is growing evidence that the mechanism by which caffeine acts to
improve athletic performance is its ability to act as an adenosine antagonist, inhibiting the
negative effects that adenosine induces on neurotransmission, excitation and pain
perception [57,61]. Current ingesting doses of caffeine ranging from 3 to 6 mg/kg,
approximately 60 to 15 minutes before exercise will provide these improvements
[57,61,62], than will last no longer than 30-45 minutes. The use of caffeine is not
recommended for all athletes/training conditions as may contribute to over-excitabilit y,
aggressiveness, increased diuresis and trend to dehydration and difficulty to sleep.

Creatine monohydrate
Creatine monohydrate is one of the most frequently supplements used by athletes [63–

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65], despite there is quite a controversy about its efficacy. Creatine is produced by the
organism and very abundant in foods containing animal protein, including fish, meat,
eggs and diaries, since is not difficult to supply even the increased demand of athletes.

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However, the everyday more frequent, restrictive diets, especially vegans and
vegetarians, are highly recommended to take creatine as part of their nutritional plan [35],
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as their muscle creatine stores will quickly shrunk. Creatine supplementation increases
intramuscular phospho-creatine stores which may have a beneficial effect on high-energy
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phosphate metabolism in High-Intensity Interval Training (HIIT) mode [35], although not
all studies found positive effects. In general creatine monohydrate improved aerobic and
anaerobic performance, strength, reduced muscle damage, and anabolic-catabolic
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hormones in male but not in female athletes [63]. Conclusive research is highly
demanded.
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Taurine
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Taurine is synthesized from the amino acid cysteine, but most of the taurine in the body
is obtained through dietary sources [66,67]. Taurine is used as a nutritional supplement
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for its anti-inflammatory and antioxidant effect. Taurine may exert an ergogenic effect by
increasing the post-exercise oxidation of lipids in specific conditions as fasting subjects
[66]. The potential benefits of taurine on muscle function may be due to its cytoprotective
effect and modulate mitochondrial function in skeletal muscle [67].

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Ketogenic diet or ketone body supplementation
Ketogenic diets have been proposed as a promising strategy to treat obesity. Specially a
very-low-calorie ketogenic diet (VLCKD) is able to induce a reduction of 20 kg of body
weight in 6 months and maintain the weight loss until 24 months [68]. This weight loss
is induced as expenses of body fat and preserving muscle mass and function [69]. In fact,
this kind of diet is able to modulate the secretion of myokines in a different way than
bariatric surgery or low-calorie diet [70,71]. Beyond the use of ketogenic diets for weight
and body composition management, these kind of diets or ketone body supplements are
also gaining interest among athletes because their ergogenic properties [72].
In this regards the supplementation with β-hydroxy-β-methylbutyrate (HMB) is currently
very common among athletes [73]. It is a metabolite of the amino acid leucine [38,63]

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and belongs to the ketone bodies group of molecules, which includes the more abundant
acetone and acetoacetate. The main role of HMB is likely serve as alternative fuel in
starving conditions. Leucine, with isoleucine and valine constitutes the group of branched

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aminoacids (BCAA), used in different ways by the muscle metabolism to provide
supplementary energy. Therefore, HMB may be a link between aminoacid metabolism
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and ketone bodies, thus being a relevant molecule in fuelling metabolism. Leucine has
been shown to stimulate muscle protein synthesis and HMB as well. Some studies showed
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that HMB could enhance sports performance in terms of aerobic power and capacity,
anaerobic capacity, strength, body composition, muscle damage, and hormone status [64].
HMB also stimulates protein synthesis in muscle via mTOR. However, these effects are
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more prominent in non-trained individuals, having low effectiveness in trained athletes.

On the other hand, HMB is a direct precursor of cholesterol synthesis [74]. It has been
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documented that supplementing with 1.5 to 3 g/day of HMB with calcium during
resistance training can increase muscle mass and strength particularly among untrained
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subjects initiating training [38]. However, there is still not sufficient scientific evidence
to warrant recommendation of consuming exogenous ketone supplements to enhance
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physical performance [75,76].

Sport Supplements and Microbiota

The composition of the diet and the microorganisms present in certain foods (for example
probiotics) in athletes can influence the composition of the intestinal microbiota with
microorganisms capable of improving energy metabolism, oxidative stress and
inflammation. Therefore, the use of sports supplements that exert a certain influence on

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the microbiota is nowadays a study objective [77]. Sports supplements that have strong
scientific evidence of interactions with microbiota are polyphenols, probiotics, protein
[78] and dietary fiber [79].
High intensity exercise can lead to a sharp increase in reactive oxygen species, causing
oxidative damage, muscle weakness and fatigue. The use of antioxidant supplements
(polyphenols) can counteract these harmful effects, in addition to acting as prebiotics and
bactericides by positively modulating the microbiota [78]. However, it has been also
described that several adaptations of skeletal muscle exercise require a moderate oxidant
concentration which may be blunted by the use of antioxidants and therefore compromise
training adaptations and performance [80]. This controversial guarantee the need of
biomarkers for personalized indications. The administration of certain strains of

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probiotics reduces the gastrointestinal disturbance [79,81] that usually occurs in elite
athletes [82], improves nutrient absorption, recovery and sports performance [79]. In
relation to protein intake, the amino acid composition has different effects on the

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intestinal microbiota, where previous research says that plant-based proteins are more
useful than animal-derived proteins [78]. Also, carbohydrates have been widely studied
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for their already recognized effect on the intestinal microbiota, and especially a higher
intake of dietary fiber is associated with greater microbial richness and diversity [79].
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EPIGENETIC MECHANISMS AS THE LINK BETWEEN FOOD

COMPONENTES AND IMPROVEMENT IN EXERCISE PERFORMANCE
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In spite of the growing interest in dietary supplementation of sportspeople to improve the

exercise performance and the increased number of scientific evidences on potential
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beneficial effects, the molecular mechanism involved in this effect is still few studied.
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Both exercise per se and ergogenic supplementations are able to induce a modulation in
gene expression and this modulation can be mediated by epigenetic mechanisms (Figure
1). In fact, within nutritional epigenetics, studies based on dietary supplementation with
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different vitamins such as folate, choline or betaine show a constant increase in DNA
methylation, as result of their condition as donors of methyl groups. Other B vitamins,
especially B2, B6 and B12, also seems to increase DNA methylation due to their role as
cofactors in the methylation process [83,84]. From all the precursors obtained from diet,
cholecalciferols (vitamin D), which action mechanism includes direct effect in the
modulation of gene´s transcription interacting with estrogens or thyroid hormones,

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potentially have larger capacities to modulate epigenetics. In fact, the effect of vitamin
D3 in bone growth and remodelling involve epigenetic mechanisms [85]. Circulating
levels of vitamin D3 were related to the methylation profile of genes related to
carcinogenesis, such as SFRP2 in colorectal cancer [86,87]. A large number of athletes
have suboptimal levels of vitamin D3 (<30 mg/dL), thus supplementation is frequently
needed [88]. This molecule and its metabolites are essential for skeletal muscle growth
and energy metabolism in mitochondria [89,90], immune function and inflammation [91],
nervous system [92], cardiovascular function [93] and lungs [94]. All together might
greatly affect athletic performance and health.
Regarding the effects of a supplementation with certain microelements such as zinc (Zn),
a variable response on methylation in specific genes was observed, so that no specific

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effects on the epigenome are generally concluded [95]. These data provide relevant
information regarding the effect on the levels of DNA methylation that these nutrients
give when they are included in the diet. Thus, the use of these components in

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supplementation products for high-performance athletes would be an effective measure
in those conditions in which a global increase in methylation resulted in direct benefits
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on performance, improved potency during exercise or even a rapid functional recovery in
athletes (Figure 1). Although the epigenetic modifications responsible for the metabolic
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and physiological adaptations induced by sodium bicarbonate, beta-alanine, nitrate,

caffeine or creatine are still unknown it is possible that their effect is mediated by
epigenetic mechanisms [96,97].
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Regarding to other nutritional strategies to maximize exercise performance and

adaptation, new data suggests that ketosis coordinates cellular function through
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epigenetic regulation, since ketosis has been associated with covalent modifications such
as acetylation and methylation [98]. Determine whether these changes in the epigenome
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are attributable to the ketone bodies themselves or to other aspects of the ketotic states
[98] is relevant topic. Elucidating the epigenetic mechanisms through which ketogenic
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diet or ketone supplements exert their action on increasing sports performance would
allow the use of ketone bodies to be guaranteed as ergogenic aids. Furthermore, including
in the diet of athletes foods providing a healthy profile of fats may have beneficial effects
on athletic performance, and these effects may be due to changes in DNA methylation
after supplementation. In this context, it has been observed that n-3 polyunsaturated fatty
acids increase methylation and extra virgin olive oil decreases the methylation of CpG
sites in the IL6 and TNF promoter region [47].

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On the other hand, the diet-related microbiota is also sensitive to homeostatic and
physiological changes associated with training. It has been shown that exercise increases
the diversity of the microbiota, thereby improving the metabolic profile and immune
responses [99]. Intense resistance exercise requires adequate measures to counteract
oxidative stress, intestinal permeability, systemic inflammation and immune responses.
The microbiota could be an important tool to improve health, performance and energy
availability, while controlling inflammation and redox levels in endurance athletes. This
effect could also be mediated by epigenetic mechanisms since there is a cross-talk
between microbiota and epigenome in the organisms that is involved in healthy promotion
[100,101]. In fact, epigenetic marks such as DNA methylation and histone modifications
are regulated in part by enzymes whose activity depends on the microbiota [101].

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Finally, the exercise itself causes a decrease in the methylation of many genes, although
in some genes the levels of methylation increase with physical activity. Physical training
can modulate the epigenome of an athlete in a favourable way towards greater

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performance; however, current research lines do not provide enough data [102]. Recently,
progress has been made towards a better understanding of the "epigenetic response" to
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specific training stimuli. It is known that both individual episodes of exercise and long-
term training programs alter epigenetic characteristics, causing specific epigenetic effects
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associated with a specific physical activity. Little it is known about differential DNA
methylation and histone modification in response to exercise, specifically in skeletal
muscle [103,104]. Therefore, the main objective of future studies will focus on
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developing specific epigenetic marker panels that allow predicting an individua l's
reaction to the stimulation of a specific training. In addition, other factors such as diet
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contribute to the modulation of epigenetic "memory", further determining this particular

reaction of a person to a specific training stimulus in the present [105]. However,
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epigenetic changes maybe more tissue-specific. This fact represents a huge challenge in
the search for epigenetic biomarkers for outcomes of tissues inaccessible without surgery.
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In this regards, several studies have demonstrated that non-invasive samples such as
leukocytes, serum or saliva could be a surrogate of target tissues for several diseases.
Thus, leukocytes reflect DNA methylation profile of adipose tissue in obesity [106,107].
Moreover, an DNA methylation mark identified in breast cancer tumors in association
with obesity [24] was also reflected in leukocytes and associated with adherence to
mediterranean diet [108]. Thus, the impact of exercise and nutrients supplementation on
DNA methylation can be also quantified in leukocytes [47] or saliva [109]. Moreover,

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other epigenetic mechanisms such as non-coding RNA, particularly miRNAs, may be
modulated by diet and exercise and these changes can be quantified in blood samples.
Circulating microRNAs (c-miRNAs) were proposed as biomarkers of exercise dose
response [110,111] or cardiac disease and inflammation signatures in response to acute
physical activity [112,113]. Therefore, peripheral blood leukocytes, serum or saliva could
be a useful tool for investigate the individual response to exercise and to evaluate the
effects of ergogenic aids to improve the exercise performance in a personalized way.
Further studies are needed to evaluate the suitability of leukocytes, serum or saliva to
provide better insight into tissue-specific epigenetic signatures in the field of exercise
performance and healthy effect. Finally, animal model experiments are also postulated as
a useful alternative in understanding the ability of exercise and diet to influence our

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epigenome. This type of study will help to elucidate the epigenetic modifications induced
by the effect of exercise in the target tissues, determinations that if carried out in humans
would involve the use of invasive techniques or surgery. Recent studies in mice focus on

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elucidating the epigenetic regulatory effect that exercise exerts on gene expression in
skeletal muscle in situations of health and disease [114,115].
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CONCLUDING REMARKS
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In summary, optimizing training, physical performance, nutrition and recovery are keys
priorities that athletes want to improve. Nutritional strategies are crucial to help athletes
perform at the highest level, so assessing the ergogenic potential of a nutritional
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supplement is important in terms of athletic performance for a given event. The increased
energy expenditure and nutrients demand maintained in time and the incremented intake
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to supply them may affect epigenetic profile. However, short-term changes in food intake
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regime, specific nutrients & food components, or ergogenic aids and supplementation are
hardly supposed to alter epigenetics in specific tissues. Very few studies have addressed
this question and little evidences are available. New studies focused on elucidating the
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epigenetic mechanisms responsible for the link between dietary supplements for athletes
and improving their performance during exercise are needed. Thus, the discovery of these
potentially ergogenic epigenetic marks will be an important advance in the elaboration of
personalized diets for sports professionals.

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Funding

The study was supported by Centro de Investigación Biomedica en Red fisiopatología de

la obesidad y nutricion (CIBEROBN) and the Miguel Servet project, initiatives of
Instituto de Salud Carlos III (ISCIII) co-financed by the European Regional Development
Fund (FEDER). PL is funded by predoctoral grant from the Instituto de Investigacion
Sanitaria de Santiago (IDIS). AC is a Miguel Servet researcher (ISCIII; CP17/0008).

Conflict of Interest Statement

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The authors declare that the research was conducted in the absence of any commercial or

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financial relationships that could be construed as a potential conflict of interest

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FIGURE LEGENDS
Figure 1. Summary of the interaction between exercise and dietary supplementation
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on athletes through the epigenetic machinery. The lifestyle of competition athletes is

marked by factors such as practicing physical activity at a high level and by a specific
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personalized diet enriched with ergogenic food supplements. To elucidate the epigenetic
mechanisms through which these factors interact and their direct consequences on the
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response, the training of athletes is currently the focus of research in this area. Therefore,
the discovery of these potentially ergogenic epigenetic marks will be an important
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advance in the development of personalized diets for sports professionals. This figure was
produced using modified elements from BioRender.io (Toronto, Ontario, Canada).

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Table 1. List of dietary supplements and nutrients to improve athletic performance and its effect potentially mediated by epigenetic mechanisms.
Epigenetic Sample
Dietary factors Potential beneficts

ro
mechanisms
Healthy diet Enhancing energy metabolism [35,38,39]. ↓ Methylation in Whole blood [116].

-p
Enhance performance and recovery [37]. CDH4, ROBO1,
↓ Fatigue [40]. PCDHGA1-12,
↑ Bioavailability of other supplements [41]. PCDHGB1-2,4-6

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genes.
↑ Methylation in

lP
DAB1, NECTIN4,
CDH9 genes [116].
Protein Maintenance of muscle mass No effects.
a
↑ Recovery after exercise [39].
rn
Healthy fat ↓ Exercise-induced inflammation [47]. ↑ ↓ Methylation in Venous blood [47].
region of IL6 and
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TNF. [47].
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Micronutrients: iron, zinc, folate, Little evidence ↑ Global DNA Primary breast tumors,
choline, betaine, B vitamins… Iron: ↑ Oxygen carrying capacity [49]. methylation levels cord blood leukocytes
[84,95,117–119]. [118,119].

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 of
Mouse fetal brain [117].

ro
Caffeine ↑ Aerobic, anaerobic or mixed performance Effect on DNA Peripheral blood cells and
[57,58,60]. methylation (e.g. leukocytes in saliva [120].

-p
↓ Pain and exertion perception ALPPL2 gene)
Improvement of muscle relaxation time [120].

re
Improvement of muscle function [35].
Creatine monohydrate ↑ Aerobic and anaerobic performance ↓ Global DNA- 5- methylcytosine

lP
↑ Strength methylation levels concentrations in liver
↓ Muscle damage [121]. tissue [121].
Improvement of hormonal status [63].
a
Taurine ↑ Physical performance [66] No effects.
↓ Exercise-related muscle damage [67]
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↑ Lipids oxidation [66]
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β-hydroxyβ-methylbutyrate ↑ Aerobic and anaerobic capacity Epigenetic Animal brain tissues

↑ Strength regulation: [122].
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↓ Muscle damage histone acetylation, Rat hippocampal tissue

Improvement of hormonal status [64]. [123].

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histone
phosphorylation

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(e.g. BDNF gene)
[122],
global DNA

-p
methylation [123].
Vitamin D3 system ↑ Aerobic and anaerobic capacity Osteoblast- IDG-SW3 cell culture and

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↑ Mitochondria energy supply osteoclast bone animal tissues [85].
↑ Nervous system capacity growth and

lP
↑ Immune response remodeling.
↓ Muscle damage Global epigenetic
↓ Inflammation [51,52,88,89] histone
a
modifications [85].
rn
Exercise + diet - miRNA modulation Serum [110].
[110].
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