Nutrients. 2016 Jul; 8(7): 441.
Published online 2016 Jul 22. doi: 10.3390/nu8070441
PMCID: PMC4963917
Abstract
This
study investigated Montmorency tart cherry concentrate (MC)
supplementation on markers of recovery following prolonged, intermittent
sprint activity. Sixteen semi-professional, male soccer players, who
had dietary restrictions imposed for the duration of the study, were
divided into two equal groups and consumed either MC or placebo (PLA)
supplementation for eight consecutive days (30 mL twice per day). On day
5, participants completed an adapted version of the Loughborough
Intermittent Shuttle Test (LISTADAPT). Maximal voluntary
isometric contraction (MVIC), 20 m Sprint, counter movement jump (CMJ),
agility and muscle soreness (DOMS) were assessed at baseline, and 24, 48
and 72 h post-exercise. Measures of inflammation (IL-1-β, IL-6, IL-8,
TNF-α, hsCRP), muscle damage (CK) and oxidative stress (LOOH) were
analysed at baseline and 1, 3, 5, 24, 48 and 72 h post-exercise.
Performance indices (MVIC, CMJ and agility) recovered faster and muscle
soreness (DOMS) ratings were lower in the MC group (p <
0.05). Additionally, the acute inflammatory response (IL-6) was
attenuated in the MC group. There were no effects for LOOH and CK. These
findings suggest MC is efficacious in accelerating recovery following
prolonged, repeat sprint activity, such as soccer and rugby, and lends
further evidence that polyphenol-rich foods like MC are effective in
accelerating recovery following various types of strenuous exercise.
Keywords: recovery, strenuous exercise, muscle damage, prunus cerasus, functional foods
1. Introduction
Prolonged, field-based, intermittent sprint sports are popular across the world at both elite and recreational levels [1].
Sports, such as soccer, field hockey and rugby, require a high volume
of energy turnover and eccentric muscle actions resulting in metabolic
and mechanically induced stress. Indeed, soccer play results in elevated
post-match inflammatory [2,3], oxidative stress [2,4,5] and muscle damage [2,4] markers, as well as decrements in physical performance [2,4,5].
The Loughborough Intermittent Shuttle Test (LIST), which closely
simulates activity patterns, and the physiological and metabolic demands
of soccer [6], has been shown to incur similar stress responses [7,8,9],
and, thus, provides a tool to induce similar physiological stress to
game-play, but in a controlled environment. In light that athlete
schedules require training or competition on multiple occasions within a
few days, the importance of recovery strategies when preparing for the
next game or training session is critical [10]. This is especially pertinent given that recovery is incomplete at 48 h following actual [2,11] and simulated [9] match-play.
Montmorency tart cherries have been shown on numerous occasions to be of benefit in exercise recovery [12,13,14,15,16],
which have been proposed to be as a result of the high concentrations
of phytochemicals, and in particular, the flavanoids anthocyanins [14,17,18,19].
These compounds can reduce oxidative stress and been shown to be a
cyclooxygenase inhibitor (COX), to a similar extent as NSAIDs [20,21]. A series of studies have investigated the use of Montmorency cherries in influencing recovery from running [14,22], heavy eccentric contractions [15,16] and cycling [12,13].
Collectively, these lines of investigation suggest that Montmorency
cherries could also be applied to aiding recovery following high
intensity concurrent sports that incorporate and very high metabolic
component that is accompanied by high intensity eccentric contractions.
Given
that Montmorency cherries have been shown to be of benefit in exercise
recovery following high intensity eccentric contractions and
metabolically challenging exercise, it makes the expectation tenable
that it could be applied to sports of a concurrent nature. Therefore the
aim of this study was to investigate the effect of Montmorency cherries
on recovery indices following a protocol designed to replicate the
physiological demands of prolonged intermittent sprint activity such as
those seen in field based sports. It was hypothesised that MC
supplementation would attenuate post-exercise inflammatory and oxidative
stress responses, and aid the return of functional performance.
2. Methods
2.1. Participants
Sixteen
semi-professional (step 5 and above in the Football Association
National pyramid, UK), male soccer players (mean ± SD age, height, mass,
predicted V̇O2max was 25 ± 4 years; 180.8 ± 7.4 cm, 81.9 ± 6.6 kg, 54.9 mL·kg−1·min−1,
respectively) volunteered to take part in the study. All procedures
were granted Ethical clearance by the University’s Research Ethics
Committee prior to testing and were conducted in accordance with the
Helsinki Declaration. Inclusion criteria required participants to have
trained in soccer consistently across the preceding 3 years and be free
of any lower limb injury for the preceding 6 months. This was assessed
through the completion of training history and health screening
questionnaires. Following both verbal and written briefings on the
requirements of the study, written informed consent was collected from
all participants.
2.2. Study Design
A
double blind, placebo controlled design with independent groups design
was employed. Participants attended the laboratory on six separate
occasions across a period of no longer than 15 days (Figure 1). On visit 1, participants completed a multi-stage shuttle test [23] in order to predict VO2max,
which was followed by familiarisation with a battery of functional
performance tests and one 15 min section of the Loughborough
Intermittent Shuttle Test Part A (LIST) [6].
Participants were then randomly but equally assigned to either a
Montmorency cherry (MC) or placebo (PLA) group, matched by predicted VO2max score (54.3 vs. 55.4 mL·kg−1·min−1).
Participants
returned to the laboratory for visit 2 within 5 days to complete the
battery of baseline functional measures that followed a standardised
warm up; these were countermovement jump height (CMJ), 20 m sprint time
(20 m), MVIC of the knee extensors, agility (5-0-5), which were preceded
by assessment of active muscle soreness (DOMS). Participants were then
provided with 24 MC or PLA beverages in a double blind manner along with
verbal and written instructions on how to consume the beverages. They
were also provided with a diet record diary and a list of foods to avoid
throughout the 4 days prior to and during the trial period. During the 4
day period leading up to the trial day, participants were contacted and
instructed to begin supplementation and dietary restrictions.
Visits
3–6 commenced at 8:00 a.m. in order to account for diurnal variation.
On visit 3, participants were required to complete an adapted version of
the LIST (LISTADAPT). Following the standardised warm-up,
participants completed a series of 12 × 20 m sprints with a 10 m
‘stopping zone’, departing every 60 s. This addition was included
because (1) the LIST protocol does not account for the many bounding,
leaping and directional changes that are associated with team sports
play; and (2) previous work from our laboratory has shown only moderate
responses with regards to the magnitude of stress response following the
LIST [9].
A secondary adaptation to the LIST protocol was the completion of 6 ×
15 min sections from the LIST Part A, as opposed to 5 × 15 min sections
detailed in the original protocol [6].
This section was included to standardise the distance covered by the
two groups (in the original protocol, LIST Part B required a run to
exhaustion, potentially resulting in group differences). During the LISTADAPT participants were provided with water ad libitum.
Visits
4–6 took place at 24, 48 and 72 h following the start of visit 3 and
required participants to complete the functional performance test
battery outlined in visit 2. Venous blood samples were collected at
baseline (prior to muscle soreness and performance test), immediately
pre-trial, immediately post-trial and 1, 3, 5, 24, 48 and 72 h
post-trial for markers of inflammation, oxidative stress and muscle
damage.
2.3. Supplementation
The
MC or PLA supplementation was provided to participants after the
initial visit along with instructions detailing the dosing schedule (30
mL twice per day, (8:00 a.m., 6:00 p.m.), 7 consecutive days (4 days
pre- and on each trial day [13])).
Supplements were prepared by mixing each dose with 100 mL of water
prior to consumption. The MC was a commercially available Montmorency
cherry concentrate (CherryActive, Sunbury, UK); previous work from our
laboratory has shown that the MC used in this study contains a total
anthocyanin content of 73.5 mg·L−1 of cyanidin-3-glucoside, a total phenolic content of 178.8 gallic acid equivalent·L−1 and an antioxidant capacity (TEAC) of 0.58 trolox equivalents·L−1 [24].
A commercially available, less than 5% fruit, cordial, mixed with water
and maltodextrin (MyProtein Ltd, Northwich, UK) until matched for
energy content of the MC (102 kcal) was used for the PLA supplement. All
supplements were prepared by an independent member of the department
prepared in opaque bottles in order to maintain the double blind design.
2.4. Dietary and Exercise Restrictions
Participants
were instructed to follow a low polyphenolic diet in the 48 h prior to
the beginning of each MC or PLA supplementation and throughout the
experimental phase of each study. A list of foods to avoid was provided
and compliance was assessed through the completion of daily food diaries
which has been successfully implemented in previous research [25].
This control measure was used to provide a washout period of
polyphenols to enable the efficacy of the phenolic-rich cherry
concentrate intervention. In addition, participants were instructed to
abstain from any exercise that was not a part of the protocol,
throughout the same time periods. Lastly, participants attended all
exercise trials following an overnight fast. These measures ensured that
dependant variable changes from baseline were likely to be in response
to the supplementation and the exercise trials implemented within each
study.
2.5. Functional Performance Tests
The
functional performance test battery was performed in the following
order on each occasion: Active muscle soreness assessment (DOMS), 20 m
sprint, 5-0-5 agility (CV 2.8% [26]),
countermovement jump (CV 1.9%), knee extensors (repeatability in
Chapter 3.4). Timings were kept consistent throughout all functional
performance tests, each test was performed 3 times (excluding DOMS),
with a 1 min rest between repetitions and 3 min rest between tests.
Delayed
onset of muscle soreness (DOMS) in the lower limbs was assessed using a
200 mm visual analogue scale (VAS) with ‘no soreness’ at one end and
‘unbearably painful’ at the other. On each occasion the VAS was used,
participants were instructed to place their hands on hips, squat down to
~90°, before standing up and immediately making a mark on the scale
consistent with their perceived soreness.
Sprint performance (20 m; coefficient of variation (CV) 0.9%) and the 5-0-5 agility test [27,28]
(CV 2.8%) were assessed using wireless telemetry and infra-red timing
gates (Brower Timing Systems, Draper, UT, USA) on an indoor athletics
track. Countermovement jump height (CV 1.9%) was assessed using a jump
mat (Just Jump, Probotics Inc., Huntsville, AL, USA); participants were
instructed to stand on the jump mat with their feet parallel and
approximately shoulder width apart. Following this, participants
completed a maximal vertical jump whilst maintaining hands on hips
through flight and landing. MVIC of the dominant knee extensors was
determined using a strain gauge (MIE Medical Research Ltd., Leeds, UK)
using the methods described previously [13]. The peak performance from each trial was used for data analysis.
2.6. Blood Sampling
Blood
samples (35 mL) were collected from a forearm vein located in the
antecubital fossa region in order to assess for markers of muscle damage
(creatine kinase [CK]), inflammation (interleukin-1-beta (IL-1-β),
interleukin-6 (IL-6), tumour necrosis factor-alpha (TNF-α),
high-sensitivity C-reactive protein (hsCRP)) and oxidative stress (lipid
hydroperoxides (LOOH)) using previously described methods [12,13];
intra-sample CVs ranged from 0.7% to 6.8%. Samples were collected into
serum gel, ethylenediaminetetraacetic acid (EDTA) or sodium heparin
treated tubes (Vacutainer®BD UK Ltd., Oxford, UK). Samples
were then immediately centrifuged (Allegra X-22 Centrifuge, Beckman
Coulter, Bucks, UK) at 2400× g at 4 °C for 15 min before having
the supernatant removed and stored in aliquots. Aliquots were then
immediately stored at −80 °C and subsequently analysed for the
respective indices in each study.
2.7. Statistical Analysis
All
data analyses were conducted using IBM SPSS Statistics 20 for Windows
(Surrey, UK) and are reported as mean ± standard deviation. All data was
confirmed as parametric via a Shapiro-Wilk test for normality.
Differences between blood marker variables were analysed by using a
group (MC vs. PLA) by time-point (Pre-supplement, Post-supplement, 1, 3,
5, 24, 48 and 72 h) mixed model ANOVA. Functional performance measures
were analysed using the same model, however with four fewer levels
(Pre-supplement, Post-supplement, 24, 48 and 72 h). Where significant
group baseline differences were apparent (MVIC, CMJ, 5-0-5 agility, 20 m
sprint, DOMS) results were normalised to baseline values prior to
subsequent statistical analysis. Mauchley’s Test of Sphericity was used
to assess homogeneity of data and where violations were present,
Greenhouse-Geiser adjustments were made. Where necessary, interaction
effects were assessed using LSD post hoc analysis. Prior to all
analyses, a significance level of p < 0.05 was set.
3. Results
MVIC (Figure 2) showed significant time (F(1,4) = 6.586, p = 0.001, η2 = 0.320), group (F(1,2) = 19.445, p = 0.001, η2 = 0.582) and interaction (F(1,4) = 8.970, p < 0.001, η2
= 0.391) effects when data was normalised to baseline values. The
decline in MIVC performance was not evident in the MC group whereas as
function had not returned to basal levels at 72 h in the PLA. The peak
difference occurred at 48 h where MVIC scores in the MC group were found
to be 19% higher.
Changes in maximum voluntary isometric contraction (MVIC; Panel (A)) and delayed onset muscle soreness (DOMS; Panel (B)) in response to Montmorency cherry (MC) or placebo (PLA) supplementation. * Group effect; $ Interaction effect (p < 0.05).
When data was normalised to baseline values, CMJ also showed significant time (F(1,4) = 30.320, p < 0.001, η2 = 0.684), group (F(1,2) = 7.336, p = 0.017, η2 = 0.345) and interaction (F(1,4) = 3.334, p = 0.028, η2 = 0.193) effects (Figure 3).
Both MC and PLA groups demonstrated reduced CMJ (vs. baseline) in the
72 h post-trial period, although the CMJ decrease in the MC group was
significantly attenuated at 24 h (5%, p = 0.022) and 48 h (6%, p = 0.017) versus placebo. Significant time (F(1,4) = 12.988, p < 0.001, η2 = 0.481) and group (F(1,2) = 7.963, p = 0.015, η2
= 0.355) effects were found for the 5-0-5 agility test. MC times for
the 5-0-5 agility were on average 3% faster (vs. PLA) across the 72 h
post-trial testing period. For the last of the performance measures, 20 m
sprint time, significant time (F(1,4) = 9.681, p = 0.001, η2 = 0.409) and interaction (F(1,4) = 3.145, p = 0.035, η2
= 0.183) effects were apparent, with both MC and PLA groups
demonstrating slower times in all three post-trial tests. At 48 h in the
MC group, 20 m sprint times were significantly (p = 0.043) faster (4%) than PLA. DOMS was significantly increased in both groups across the 72 h post-trial period (F(1,4) = 37.206, p < 0.001). A significant group effect (F(1,2) = 8.486, p = 0.011, η2 = 0.377) showed that DOMS ratings were lower in the MC group (vs. PLA), which were mirrored by interaction effects (F(1,4) = 4.069, p = 0.013, η2 = 0.225) at 24 (p = 0.044), 48 (p = 0.018) and 72 h (p = 0.007), which showed almost complete recovery of at 72 h.
Comparison
of countermovement jump (CMJ) height with Montmorency cherries (MC) or
placebo (PLA) supplementation. * Group effect; $ Interaction effect (p < 0.05).
With regards to inflammatory markers, IL-6 (Figure 4) was found to be elevated in both groups following the trial (F(1,8) = 52.180, p < 0.001, η2 = 0.788). Group comparisons (F(1,2) = 10.223, p = 0.006, η2
= 0.422) demonstrated an overall significantly attenuated IL-6 response
to the trial in MC (vs. PLA), with significant interaction effects (F(1,8) = 3.313, p = 0.003, η2 = 0.191) showing peak differences of 3.10 pg·mL−1 occurring immediately post-exercise (p = 0.03). Further inflammatory marker data for plasma IL-8 (F(1,8) = 4.905, p = 0.010, η2 = 0.259), TNF-α (F(1,8) = 6.343, p < 0.001, η2 = 0.312) and hsCRP (F(1,8) = 20.298, p < 0.001, η2
= 0.592) revealed significant increases in each variable in the 72 h
following the trial, however, group and interaction comparisons failed
to identify differences. IL-1-β was not found to be increased at any
measurement point across the trial period. The trial significantly
increased CK in both groups (F(1,4) = 10.243, p = 0.004, η2 = 0.423), although no group or interaction effects were found. Peaks values of 1200 IU/L were attained at 24 h (Table 1).
Interleukin-6
(IL-6) responses in the Montmorency cherry (MC) and placebo (PLA)
groups to the adapted Loughborough Intermittent Shuttle Test (LISTADAPT) exercise. * Group effect; $ Interaction effect (p < 0.05).
Lipid hydroperoxides were increased in the 72 h post-exercise period as indicated by a significant time effect (F(1,8) = 5.973, p < 0.001, η2
= 0.289). Peak increases of 35% above baseline occurred at 5 h.
Although there was a tendency for higher PLA group values, no
significant group or interaction effects were found. A summary of
variables is reported in Table 1.
In order to identify any group differences in LISTADAPT performance, a comparison of sprint times during the LISTADAPT protocol was performed using Student’s T-test. No group differences in LISTADAPT sprint performance were found (t = 1.511, p = 0.153).
4. Discussion
The
main finding of this study was that participants supplemented with MC
were able to maintain greater functional performance than PLA
counterparts following prolonged intermittent sprint activity. More
specifically, MVIC, CMJ, 20 m sprint and 5-0-5 agility performances were
superior in the 72 h post-exercise with MC (vs. PLA). In addition, DOMS
and IL-6 were lower in the MC group throughout the post-trial period.
The
attenuated declines in muscle performance are consistent with the
findings of previous studies investigating MC as a recovery aid [12,13,14,15,16] and additionally the magnitude of MVIC decline following the LISTADAPT was comparative to previous work utilising a similar protocol [29].
The MVIC performance was on average 17%, superior in the MC group (vs.
PLA). MC supplementation also resulted in better (vs. PLA) CMJ
performance at 24 and 48 h. Sprint times in both groups were slower in
the 72 h post-exercise period, however at 48 h the MC group was
significantly faster. Agility (5-0-5) times were also faster in the MC
group by an average of 3% across the 72 h post-trial period.
Interestingly, the MVIC results in the 72 h following exercise suggests
MC supplementation abolishes declines in this performance measure—a
result that has been previously reported [13].
These
data support the idea that supplementation with MC protect declines in
muscle function following strenuous exercise, specifically in activity
akin to repeated sprint sports and games play such as rugby, soccer and
field hockey. A reduction in post-exercise IL-6 suggests a lower acute
inflammatory response to the exercise bout that might contribute to the
performance differences between groups. The COX, prostaglandin, IL-6
pathway, which are activated during the secondary inflammatory response
to cellular disruption, has been associated with proteolytic and
lipolytic processes [30]
and subsequently muscular performance could be inhibited. Seemingly, MC
supplementation reduced (but did not abolish) this process and allowed
for greater maintenance of muscular performance in the recovery period.
Conversely, there were no group differences in hsCRP. This is unexpected
given that IL-6 is implicated as a signalling molecule for the
expression of hsCRP [31,32].
We are unable to resolve the discrepancy between IL-6 and hsCRP and
therefore suggest further work is needed to identify the mechanism by
which MC might exert its anti-inflammatory responses in response to
strenuous exercise.
In contrast to previous work [12]
that examined repeated days cycling exercise, there were no differences
in LOOH between groups. The obvious discrepancy between study findings
may be attributed to single versus repeated days exercise. Unlike Bell
et al. [12],
where repeated days cycling exercise (metabolic challenge) were used,
the current study investigated a single bout of strenuous exercise that
incorporated both a metabolic and mechanical exercise stress.
Conceptually, the accumulated stress response from repeated day’s
exercise would be greater than a single bout, but of course the
modalities, cycling versus simulated concurrent exercise, pose very
different exercise challenges and hence the redox response may also
differ considerably between exercise stimuli.
In agreement with previous work [16,22],
the lower post-exercise DOMS in the MC group provide further evidence
for the protective effect of cherries. Despite this, previous work from
our lab [13]
using MC has not shown this positive effect. This discrepancy may be
attributed to the different modes of exercise employed to induce stress.
Whilst muscle actions during cycling are almost exclusively concentric [33], the repeated sprints and decelerations during the LISTADAPT
protocol in the present study, place a heavy eccentric load on the same
muscle groups and as a result are likely to incur greater mechanical
stress. Indeed, DOMS ratings from the present study were consistently
higher than those in the cycling studies [12,13].
In further support of this supposition, CK (an index of cellular
disruption following damaging exercise) was considerably higher than the
aforementioned cycling studies. However, the CK response reported in
previous work [14,15] using protocols that also incorporate a heavy eccentric component, showed no evidence for a protective effect of MC.
The
high-intensity, prolonged, intermittent nature of soccer and other
repeated sprint sports places a high degree of both mechanical and
metabolic stress [2],
which is reflected by the increase in the appearance of physiological
stress responses in the present study. Whilst this is not the first
study to demonstrate accelerated recovery of functional performance with
MC supplementation, it is the first to do so following simulated games
play and therefore represents an important wider application of this
intervention to aid exercise recovery in sports of an intermittent
sprint nature, such as soccer, rugby, field hockey and basketball.
Collectively, there is a growing body of evidence suggesting that MC has
the ability to facilitate exercise recovery—perhaps by modulating
inflammation and/or oxidative stress. The exact mechanisms behind these
promising data are not clear, so it seems prudent to explore further;
perhaps by using animal, cellular and molecular techniques to provide a
greater understanding of the application of MC and other phenolic-rich
foods.
In summary, this study provides
further evidence for the use of MC as a recovery aid. For the first
time, MC supplementation has been shown to accelerate the recovery of a
number of functional performance measures following prolonged
intermittent sprint activity and suggest that some dampening of the
post-exercise inflammatory processes might be responsible. With regards
to application, the dampening of such responses could be highly
advantageous in sports requiring athletes to complete high volumes of
training whilst ‘in-season’, or athletes competing in tournament
scenario’s that require multiple performances within a short time period
and the ability to recover in sufficient time is a challenge.
Additionally, although dietary restrictions were imposed throughout the
study period, the results suggest that sports requiring sprinting or
high intensity directional changes might benefit from MC supplementation
when playing schedules are congested and recovery time is limited
between games. Finally, the issue of modulating the post-exercise
oxidative stress and inflammatory response has raised concerns; insofar
as these stressors are implicated in the adaptive response. Although
there is no evidence that the adaptive response is affected by
functional foods, this question should be addressed in order to
determine if periodiation of these sorts of supplements is warranted.
Author Contributions
P.G.B.,
E.S. and G.H. conceived and designed the experiments; P.G.B. performed
the experiments; P.G.B., G.W.D. and G.H. analyzed the data; G.W.D.
contributed reagents/materials/analysis tools; P.G.B., E.S. and G.H.
wrote the paper.
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