Advances in Physical Education
2013. Vol.3, No.4, 197-204
Published Online November 2013 in SciRes (
Open Access 197
Effects of Carnosine and Beta-Alanine Ingestion on Anaerobic
Sprint Performance and Peripheral Blood Mononuclear Cell
Interleukin-6 and -10 Gene Expression
Pietro Luigi Invernizzi1, Bruno Venerando2, Francesco Di Pierro3,
Sandro Saronni4, Nadia Papini2
1Department of Biomedical Science for Health, University of Milan, Milan, Italy
2Department of Medical Biotechnology and Translational Medicine, University of Milan, Milan, Italy
3Velleja Research, Pontenure, Piacenza, Italy
4School of Sports Science, University of Milan, Milan, Italy
Received September 25th, 2013; revised October 25th, 2013; accepted November 2nd, 2013
Copyright © 2013 Pietro Luigi Invernizzi et al. This is an open access article distributed under the Creative
Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium,
provided the original work is properly cited.
Chronic administration of β-alanine has been shown to increase muscle carnosine content and improve
anaerobic performance. It is not clear whether acute ingestion of carnosine and beta alanine may have the
same effects. With a view to investigating acute effects of carnosine and β-alanine ingestion on anaerobic
intermittent running performance and on the responses of Interleukin-6 and -10 to exercise, twelve
healthy, young, active participants (age: 21 ± 4 years) underwent the running-based anaerobic test (RAST)
twice (with 30 min recovery in between) on two separate occasions (randomized, crossover design). The
test consisted of 6 × 35-m sprints interspersed with 10 s rests after acute ingestion (4 hours before the test)
of either 2 g L-carnosine + 2 g β-alanine or placebo. The overall performance decreased (RAST1 vs
RAST2, carnosine + β-alanine: 32.8 ± 1.3 s, 33.4 ± 1.2 s; Placebo: 32.9 ± 1.0 s, 33.6 ± 1.2 s), pain after
RASTs increased (RAST1 vs RAST2, carnosine + β-alanine: 3.0 ± 2.1 a.u., 4.2 ± 1.9 a.u.; Placebo: 3.0 ±
1.8 a.u., 3.4 ± 1.2 a.u.) almost in the same way in both groups, and RPE did not show any difference. IL6
and IL10 gene expression increased and decreased respectively in response to exercise in the same fash-
ion in both conditions. During RAST 2 we found a potentially increased performance in the carnosine +
β-alanine group (main effect of condition, p < 0.05). In conclusion these findings suggest that acute ad-
ministration of carnosine + β-alanine does not influence the cytokine response to exercise but might have
a very small enhancing effect on anaerobic sprint performance.
Keywords: Running; Supplementation; Cytokines
Dipeptide carnosine (β-alanyl-L-histidine) is found in vari-
ous tissues such as brain and heart, but especially in skeletal
muscle fibres (Harris et al., 2012) of both types I and II, where
carnosine concentration is the highest (Baguet et al., 2010).
Although carnosine is synthesized endogenously, its tissue
concentration is influenced by the diet (Baguet et al., 2010).
Indeed, exogenous supplementation of β-alanine (precursor and
limiting factor in the synthesis of carnosine) has been shown to
increase muscle carnosine concentration in vitro (Margolis et
al., 1985) and in animal models (Hill et al., 2007). Beta-alanine
is usually administered orally and for a period of 1 - 4 weeks
(Jagim et al., 2013), since its chronic consumption seems to be
necessary to ensure carnosine increase inside muscle fibres
(Derave et al., 2007).
Carnosine levels seem to be correlated to exercise perform-
ance: Suzuki et al (Suzuki et al., 2002) have observed a positive
correlation between carnosine content in the vastus lateralis and
the power generated at the end of a 30-second cycle ergometer
test (Wingate test) in untrained individuals.
Hill et al. (Hill et al., 2007) demonstrated that β-alanine ad-
ministration in active males produced a significant increase in
carnosine levels in skeletal muscles, which was related to an
improvement of exercise performance. In particular, β-alanine
was administered to untrained males for 10 weeks, producing
not only a remarkable increase in carnosine content in the vas-
tus lateralis, but also a significant prolongation of the time to
exhaustion during a cycling test performed at 110% of energy
production and maximal heart rate, where the resistance time is
estimated to be about 2.5 minutes (Hill et al., 2007). The role
played by carnosine, or by β-alanine, is of great interest not
only for untrained subjects: both compounds have a positive
effect on strength in trained athletes, especially sprinters, row-
ers and body-builders as well (Baguet et al., 2010; Derave et al.,
At physiological pH, carnosine exerts a strong buffering ac-
tion (stabilization of the degree of acidity) that is essential dur-
ing muscle activity, usually associated with an acidification of
the intracellular compartment (Hobson et al., 2012). Further-
more, carnosine has been recognized to have antioxidant prop-
erties (Alhamdani et al., 2007), as well as stimulating actions
on the immune system and various neurotransmitters (L-carno-
sine lowers neural activities of sympathetic nerves and en-
hances those of parasympathetic nerves) (Nagai et al., 2012).
Some evidence indicates that carnosine is available and ready
to exert its function inside the muscle fibres just after a few
hours from its ingestion in high doses (Begum et al., 2005;
Gardner et al., 1991).
Carnosine was recently shown to play a role in inflamma-
tion-reducing, pro-inflammatory cytokine release in vitro and in
vivo. In particular, a study performed in murine microglial cells
demonstrated that carnosine inhibits the synthesis of inflam-
matory mediators during lipopolysaccharide-induced inflam-
mation (Fleisher-Berkovich et al., 2009). This protective effect
of carnosine was also demonstrated in astroglial cells (Nicoletti
et al., 2007). Moreover, carnosine prevents IL8 release, follow-
ing H2O2 treatment, in human intestinal Caco-2 cells (Son et al.,
2004). Lee et al. demonstrated that the intake of carnosine sig-
nificantly suppresses the increase in inflammatory cytokines
interleukin-6 (IL6) and tumour necrosis factor-alpha (TNF-α) in
diabetic mice (Lee et al., 2005).
Several studies have provided evidence that strenuous exer-
cise induces increased expression of several pro- and anti-in-
flammatory cytokines in skeletal muscle and blood (Ostrow-
ski et al., 1999). Among these, interleukin-6 seems to play a
major role: IL6 expression is up-regulated during the exer-
cise-induced inflammatory process and exerts an anti-inflam-
matory effect leading to the release of anti-inflammatory inter-
leukins, such as interleukin-10 (IL10), and inhibiting TNF-α
production (Capomaccio et al., 2011; Fischer, 2006; Petersen &
Pedersen, 2006). Large amounts of IL6 are produced in skeletal
muscles and released into plasma in response to exercise and
training, then muscle IL6 seems to stimulate the release of IL6
from peripheral blood mononuclear cells (PBMCs) (Fischer,
2006; Rhind et al., 2001).
In spite of the potential for a positive effect of acute inges-
tion of carnosine on high-intensity exercise performance, in a
recent study (Invernizzi et al., 2013), we failed to demonstrate
any improvement in performance during an intermittent
high-intensity running test following acute ingestion (4 hours
prior to the test) of 1 g L-carnosine + 1 g β-alanine, in com-
parison with that of the placebo group. These results seem to be
in agreement with the data obtained by other authors who un-
successfully tried to improve high-intensity cycling perform-
ance following acute ingestion of a soup with a high content in
carnosine and anserine (Suzuki et al., 2006). It cannot be ex-
cluded that in these two previous studies the dose of carnosine
administered to the participants was too low (13.5 and 6.2
mg·Kg1 respectively).
The aim of this study was to evaluate whether acute ingestion
of a high dosage of L-carnosine and β-alanine (2 g + 2 g re-
spectively) would induce an improvement in performance dur-
ing a running anaerobic sprint test (RAST) (Zagatto et al., 2009)
in comparison with placebo. In addition, we investigated the
effect of acute carnosine + β-alanine supplementation on the
synthesis of exercise-induced cytokines, and in particular we
studied the mRNA expression of interleukine-6 and -10 in
PBMCs in response to an anaerobic sprint.
On the basis of our previous results, in this study we decided
to double the carnosine + β-alanine dosage (about 26.6 mg·Kg1
body weight each compound) and the physical effort (by asking
the volunteers to repeat a second RAST following a recovery
period). Our hypothesis was that this high dosage of carnosine
+ β-alanine (compared to the dosage of previous researches)
would enhance performance, both in the first and especially in
the second RAST, and affect the mRNA expression of inter-
leukine-6 and -10 in PBMCs.
The rationale of our hypothesis is based on the fact that a
high acute (4 hours after the ingestion) dosage of carnosine and
β-alanine (2 g + 2 g) can increase the muscle content of car-
nosine such that the muscle buffering capacity raises. From an
ethical point of view we decided to test the biological effects of
our hypothesis first. Once this hypothesis is tested, a biopsy
study would be reasonable and appropriate to look into the
mechanisms responsible. Although this decision represents a
limit for this study and can raise criticisms, we would like to
avoid any invasive practice for our participants prior to having
ascertained the enhancing effect on performance of an acute
administration of carnosine + β-alanine.
Materials and Methods
The participants were recruited in the Milan area in Italy,
according to the following inclusion criteria: males aged be-
tween 20 and 30 years, no smokers, absence of chronic diseases
or chronic drug treatment. The study was approved by the
Ethical Committee of the University of Milan. All the subjects
signed a written informed consent prior to participation, ac-
cording to the Declaration of Helsinki. All the procedures used
complied with the Good Clinical Practice (GCP) principles.
Twelve healthy male subjects were enrolled (age: 21 ± 4
years, height 181 ± 3 cm, body mass 75.1 ± 4.6, body fat per-
centage 12.0 ± 3.6, body mass index: 23.0 ± 1.3 kg/m2); all the
participants were on a stable diet and were requested not to
modify their diet during the period of the data collection. They
were physically active and used to exercise on average 3.3 ±
2.2 times a week for 1.5 ± 1.0 hours (carnosine + β-alanine
group) or 2.2 ± 1.2 times a week for 2.5 ± 1.3 hours (placebo
group) at vigorous and moderate intensities according to the
International Physical Activity Questionnaire (IPAQ) (Craig et
al., 2003). Most of the participants recruited for this study were
soccer players accustomed to repeated intermittent all-out
sprints. Prior to the beginning of the study, the volunteers par-
ticipated in a familiarization session with the intermittent all-
out running sprint test used in this study.
Study Desi gn
The study was conducted using a double blind, randomized,
counterbalanced, cross-over, placebo-controlled design. The
randomization and the preparation of treatment and placebo
tablets were made by a person unrelated to the study. The
treatment and placebo conditions were disclosed to the authors
of the study only after the statistical analysis had been com-
pleted. All the participants underwent two double intermittent
all-out running tests, separated by 30 min on two separate occa-
sions: on the first test day, half of the participants (selected by a
computer-assisted simple randomization procedure) ingested 2
g of L-carnosine + 2 g of β-alanine (DDM Carnosina, kindly
provided by Omeopiacenza s.r.l., Pontenure—PC, Italy) 4
hours before the test, whereas the other half of the participants
ingested placebo (made of microcrystalline cellulose, calcium
phosphate, hydroxypropyl methylcellulose, magnesium stea-
Open Access
Open Access 199
rate, silicon dioxideno and having the same appearance and
taste as the tablets made of L-carnosine and β-alanine), and vice
versa on the second test day (2 weeks later). The purity of the
supplement was independently tested by the formulator of the
tested product (Velleja Research) with the aim to exclude the
presence of performance-enhancing compounds such as ana-
bolic agents or stimulants. None of the participants reported the
presence of possible side effects following the high acute inges-
tion of L-carnosine + β-alanine neither during the testing period
nor in the next hours or the day after (in particular no one ex-
perienced any form of paresthesia that is recognised to be a
likely symptom when high dosage of carnosine are adminis-
tered). The 30 min interval between RAST 1 and RAST 2 was
chosen so as to participants could rate the session RPE also
after RAST 1.
Two weeks seems to be a more than sufficient washout pe-
riod when carnosine is acutely administered to human subjects
(Suzuki et al., 2006). In this study the participants treated with
DDM Carnosina, even if the product contains β-Alanine, are
simply called “carnosine group” or “carnosine condition”.
DDM Carnosine (a commercially-available supplement), in
agreement with the Italian law number 169/2004, was notified
to the Minister of Health in 2010 (Registration number: 53436)
and registered as food supplement with both its active ingredi-
ents (β-alanine and L-carnosine) belonging to the positive list
of ingredients admitted as nutraceuticals, and with all of its
excipients being food grade. DDM Carnosine and the placebo
tablets were both manufactured by Procemsa Farmaceutici
(Nichelino, Turin, Italy). According to the main formulator of
the tested product (DDM Carnosina), the combination of car-
nosine + β-alanine was thought with the aim to maximize the
carnosine content within the muscle cell. Theoretically, subjects
with low level of carnosinase might directly benefit of the car-
nosine as is while subjects with higher level of carnosinase
might take advantage of the fraction of beta alanine already
present in each tablet (250 mg) plus the amount coming from
the splitting up of carnosine (β-alanine and L-istidine).
The Running-Based Anaerobic Sprint Test (RAST)
A standardized 10-min warm-up, consisting of 5 min running
at a moderate pace followed by 5 accelerations over a distance
of 40 m, interspersed with 1-min passive recovery, was com-
pleted by each participant prior to the beginning of the RAST.
After 5-minute passive recovery, the participants started the
first RAST (RAST 1). A thirty-minute passive recovery was
observed before the beginning of the second RAST (RAST 2).
The RAST was performed as follows: starting from a stand-
ing position, on a 400 m track, each participant sprinted all-out
over a distance of 35 m for six times (forth and back), inter-
spersed by 10-second passive recovery. The performance times
were electronically recorded using photocells gates located 0.5
meters ahead of the start line and at a height of 1.3 meters from
the ground. The 35 m were measured as the distance between
the two photocells gates. During all the sprints the athletes were
verbally encouraged by the researchers, in order to obtain the
maximum performance. The RAST has been shown to be a
valid and reliable tool for assessing anaerobic power during
running (Zagatto et al., 2009). The outcome measures were 1)
time to complete each section of 35 m, 2) total time (the sum of
the six partial times over the 35 m sections), 3) ratings of per-
ceived exertion (RPE) and muscle pain (PAIN) immediately
after the completion of each test (RPE 1 and PAIN 1, RPE 2
and PAIN 2 after RAST 1 and RAST 2 respectively) using the
validated CR 10 Borg scale (Borg, 1998). Thirty minutes after
the end of RAST 2, RPE was assessed again. The RPE values
measured at this time were used to assess the internal load
through the session RPE-based method (Foster et al., 2001;
Impellizzeri et al., 2004).
Blood Sample Collection and RNA Extraction
One hour before the beginning of RAST 1 and one hour after
the completion of RAST 2, 200 μl of blood were drawn from
the fleshy pad of the fingertip of each subject.
Blood was collected in a heparinized tube, and total RNA
from PBMCs was isolated using the PureLink Total RNA
Blood Purification kit (Invitrogen) according to the instructions
provided by the manufacturer.
Reverse Transcription and Real-Time PCR
The iScript cDNA Synthesis kit (Bio-Rad Laboratories) was
used to reverse-transcribe 150 ng RNA, and gene expression of
IL6 and IL10 was assayed by real-time PCR.
Real-time PCR was performed by the iCycler thermal cycler
(Bio-Rad Laboratories) using cDNA corresponding to 10ng
total RNA as a template. PCR mixture included 0.2 μM primers,
50 mM KCl, 20 mM Tris/HCl pH 8.4, 0.8 mM dNTPs, 0.7U
iTaq DNA Polymerase, 3 mM MgCl2, and SYBR Green (iQ
SYBR Green Supermix from Bio-Rad Laboratories) in a final
volume of 20 μl. Amplification and real-time data acquisition
were performed using the following cycle conditions: initial
denaturation at 95˚C for 3 minutes, followed by 40 cycles of 10
seconds each at 95˚C and 30 seconds at 58˚C. The fold change
in expression of the different genes was normalized to the ex-
pression of β-actin mRNA and was calculated by the equation
2−ΔΔCt. All reactions were performed in triplicate. The primers
used are reported in Table 1. The accuracy was monitored by
the analysis of the melting curves.
Statistical Analysis
The normality of data distribution was preliminary checked
by the Shapiro-Wilk’s test. Differences in performance (total
time given by the sum of each 35 m run), perceived exertion
(RPE) and pain at the end of each RAST, between the two con-
Table 1.
Primers used for real-time PCR.
Primer Forward Reverse
ditions (carnosine and placebo, main factor Condition) and the
two RAST (RAST 1 and RAST 2, main factor Time), were
tested using a 2 × 2 (Condition × Time) fully repeated multi-
variate ANOVA measures. Session RPE (30 minutes after the
completion of RAST 2) between treatment and placebo was
tested using a 2 × 2 (Condition × Time) fully repeated multi-
variate ANOVA.
A series of 2 × 6 (Condition × Time) fully repeated multi-
variate ANOVA measures were used to test the differences
between the two conditions (treatment with carnosine or pla-
cebo, main factor condition) and the times to complete each
bout of the RAST.
Changes in the relative expression of IL6 and IL10 between
groups were calculated using a 2 × 2 (Condition × Time) fully
repeated multivariate ANOVA measures, where the main factor
Condition is represented by the carnosine or placebo admini-
strations and the main factor Time is represented by the base-
line (one hour prior to RAST 1) and post exercise (one hour
after the completion of RAST 2) blood samples. Data are pre-
sented as the means ± standard deviations (S.D.). The level of
significance was set at α < 0.05.
Performance, Perceived Exertion and Pain
No significant interactions were found for performance (p =
0.472), RPE (p = 0.653; carnosine + β-alanine: RAST 1 6.3 ±
2.0 vs RAST 2 6.8 ± 1.9; placebo: RAST 1, 6.3 ± 2.2 vs RAST
2, 6.9 ± 1.5) or pain (p = 0.099). However, the significant main
effect of time on performance (p < 0.001) and pain (p < 0.05;
carnosine + β-alanine: RAST 1, 3.0 ± 2.1 vs RAST 2, 4.2 ± 1.9;
placebo: RAST 1, 3.0 ± 1.8 vs RAST 2, 3.4 ± 1.2), showed that
between RAST 1 and RAST 2 there was a decrease in per-
formance (that is, a higher total time to complete RAST 2 in
comparison with RAST 1) (Figure 1) and an augmented per-
ceived pain at the end of the RAST 2 in comparison with RAST
1. No significant interaction and main effect of condition were
detected for session RPE; on the contrary, the significant main
effect of time (p < 0.01) showed an augmented session RPE
over time (carnosine condition, 68 ± 28 a.u. vs 239 ± 49 a.u.,
post RAST 1 vs RAST 2 respectively; placebo, 62 ± 17 a.u. vs
233 ± 52 a.u. post RAST 1 vs post RAST 2 respectively).
During RAST 1 the time to complete each 35 m bout sig-
nificantly increased in both conditions (main factor time, p <
0.001) but without significant interaction (Condition × Time, p
= 0.281) (Figure 2). Both the main effects of time (p < 0.001)
and condition (p < 0.05) of RAST 2 were found significant
(Figure 3). The time to complete each 35 m bout progressively
increased in both conditions; at group level, the significant
main effect of condition (p < 0.05) stands for an overall faster
time to complete the RAST 2 in the carnosine condition.
IL6 and IL10 Gene Expression
As shown in Figure 4 we found significant differences in the
expression of IL6 and IL10 before the beginning of RAST 1
and one hour after the completion of RAST 2. In particular, IL6
expression showed approximately a two-fold increase after
Figure 1.
Effects of DDM-C, compared to Placebo, on performance, expressed as total time to complete the Running Anaerobic Sprint Test (RAST),
between RAST 1 and RAST 2; *Significant main effect of Time, p < 0.05. DDM-C, L-carnosine + β-alanine.
Open Access
Figure 2.
Effects of DDM-C, compared to Placebo, on the time to complete each single sprint of RAST 1 (Running Anaerobic Sprint
Test); *Significant main effect of Time, p < 0.05. DDM-C, L-carnosine + β-alanine.
Figure 3.
Effects of DDM-C, compared to Placebo, on the time to complete each single sprint of RAST 2 (Running Anaerobic
Sprint Test); *Significant main effect of Time, p < 0.05; #Significant main effect of Condition, p < 0.05. DDM-C,
L-carnosine + β-alanine.
exercise (Figure 4(a)), whereas IL10 expression was reduced
by approximately fifty per cent (Figure 4(b)). However, there
were no statistically significant differences in IL6 and IL10
gene expression between the carnosine and placebo groups (p >
0.05). Discussion
The purpose of this study was to determine if an acute inges-
tion of L-carnosine and β-alanin would improve performance e
Open Access 201
Figure 4.
Interleukin-6 (a) and -10 (b) mRNA expression by real time PCR before the beginning
of RAST1 (pre) and one hour after completion of RAST 2 (post). Values are the
means ± SD. Significant main effect of Time *p < 0.05 and **p < 0.01.
during an anaerobic sprint test. Moreover, we analysed whether
this supplementation would affect exercise-induced inflamma-
tory response in blood, in particular decreasing IL6 gene ex-
pression, therefore exerting a positive effect on performance.
The main findings of this study are that ingestion of a dose of
carnosine + β-alanine (2 + 2 g), compared to a placebo con-
dition, 4 hours prior to the exercise did not affect the overall
performance or the first bout of an intermittent all-out running
exercise but had an enhancing, albeit very low, effect on the
second bout of the same intermittent exercise performed 30
minutes later. In addition, the large dose of carnosine ingested
by the participants did not affect the IL6 or IL10 response to
exercise in comparison with the placebo condition.
During RAST 1, the time to complete each 35 m bout in-
creased similarly in both conditions, with no difference in total
time. During RAST 2, the trend was similar to RAST 1 but a
likely small difference was detected between the two condi-
Contrary to our hypothesis, performance was not improved
following the high dosage of carnosine during RAST 1. Inter-
estingly, during RAST 2 there was a very small enhancement in
performance in the carnosine condition in comparison with
placebo. It is likely that this effect is very modest and this issue
is supported by the low-to-moderate partial eta squared value
(partial η2 = 0.363). However, from a practical point of view,
just few hundredths of a second can represent a sufficient gap
to make the difference between the winner and the second place
in sprinting races of different sports (such as 200 and 400 m in
the track and field, 50 and 100 m in swimming, 500 and 1000
m time trial in track cycling, etc.). We believe this is a concern
not to be underestimated.
According to Suzuki et al. (2006) the possible enhancement
in performance during RAST 2 might be due to the role played
by the non-bicarbonate buffering system. Carnosine, within the
muscle cells, can rapidly accept the protons produced during
high-intensity exercise in favour of a preservation of the
[HCO3]. The lack of improvement in high-intensity cycling
performance (Suzuki et al., 2006) was attributed to the limited
buffering potential of the carnosine supplementation (dosage
about 6.2 mg·Kg1). A higher dosage, adopted in our previous
study (data not published), failed to improve performance. In
this study a double dosage of carnosine failed again to show an
improvement in performance during RAST 1 but a possible
enhanced performance was highlighted in RAST 2. It may be
Open Access
supposed, therefore, that the carnosine buffering capacity can
be exerted when the total duration of a high-intensity intermit-
tent exercise is longer than 30 - 40 seconds. Indeed, in the pla-
cebo condition any carnosine present in the muscle might have
been depleted at the end of RAST 1 and was therefore lower
during RAST 2. Such a circumstance did not likely occur in the
carnosine condition. This hypothesis seems to be in harmony
with the consideration proposed in a recent meta-analysis (Ba-
guet et al., 2010) that shows how the effect of chronic supple-
mentation of β-alanine is particularly effective in exercises
lasting from 60 to 240 s. It can be believed that similar mecha-
nisms occur also following acute ingestion of L-carnosine +
β-alanine: in the present study the total duration of exercise is
around 60 - 65 s and the higher amount of L-carnosine and
β-alanine administered acutely to the participants might have
significantly increased the muscle content of carnosine and the
muscle buffering capacity.
A further and alternative mechanism through which muscle
carnosine can improve muscle performance (Dutka & Lamb,
2004) is the increased Ca2+ sensitivity exerted on the contractile
fibres. A high dosage of carnosine, like that used in this study,
might have had an effect on the Ca2+ sensitivity and was likely
to have contributed to lowering times in some of the sprints
during RAST 2.
In addition, due to the potential antioxidant activity of car-
nosine (Kohen et al., 1988; Decker et al., 2000), some not yet
well understood mechanisms might have contributed to posi-
tively affect short term all out performance in the carnosine
group of the present study.
As far as performance is concerned, this study is in agree-
ment with a study on cycling (Suzuki et al., 2002), in which
carnosine was supposed to have a positive effect towards the
end of a Wingate test. However, this study is in contrast with
previous results showing a lack of increase in intermittent cy-
cling performance after the ingestion of carnosine, although it
was effective in the preservation of blood [HCO3] (Suzuki et
al., 2006). On the whole, it is necessary to point to some dif-
ferences among these studies to understand the diverse and
similar results. First of all, the type of exercise: in this study we
used running while in the other studies it was cycling (Suzuki et
al., 2002; Suzuki et al., 2006). In addition, the test was based on
continuous versus intermittent cycling. Second, the amount of
carnosine administered to the subjects was higher than that used
in the two studies on cycling (Suzuki et al., 2002; Suzuki et al.,
2006) and in our previous study (data not published).
Although we observed a significant difference in interleukin
gene expression prior to, and following, exercise, we did not
find a correlation with L-carnosine-β-alanine and placebo
treatments. The marked exercise-associated increase in IL6
gene expression in PBMCs is in agreement with other studies
(Rhind et al., 2001; Sander et al., 1998; Hagiwara et al., 1995),
but supplementation failed to modify interleukin expression and
in particular the IL6/IL10 ratio. It is conceivable that an acute
supplementation may not be enough to modulate PBMC cyto-
kine production or to modify IL6 and IL10 expression, and
probably a prolonged period of supplementation could be need-
ed to have a significant anti-inflammatory effect.
One of the main limits of our study is the lack of blood
analysis, which could have been evidence of potential differ-
ences in lactate, pH or [HCO3] and, consequently, provide a
better explanation of the mechanisms underpinning the possible
positive effect of carnosine on performance.
In conclusion, the ingestion of L-carnosine and β-alanine (2
g + 2 g) 4 hours prior to an intermittent all-out anaerobic exer-
cise does not have any effect on the PBMCs interleukin-6 and
-10 gene expression and on a first trial of intermittent running
performance but increases overall performance during a second
trial run 30 min after the first one. The mechanisms of this in-
crease in performance could be ascribed to a long term en-
hanced muscle buffering capacity exerted by the acute ingestion
of L-carnosine and β-alanine.
The Authors wish to thanks all the participants for their en-
thusiasm and commitment during data collection. A particular
thank to Andrea Bosio for his valuable feedback during the
design of the study and the preparation of this manuscript.
The authors, but Di Pierro F. who is the main formulator of
the tested product (DDM Carnosina), declare no conflicts of
interest that are directly relevant to the content of this article.
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