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Research Paper
IJERSS Volume 2 | Issue 3 MAR 2015
I n t e r n a t i o n a l J o u r n a l o f E n g i n e e r i n g R e s e a r c h a n d S p o r t s S c i e n c e
Page
1
Foong Kiew Ooi
Sports Science Unit
,
S
chool of Medical Sciences, Universiti Sains Malaysia
,
16150, Kubang Kerian,
Kelantan Malaysia
Mohamed Husna
Afifah
Sports Science Unit
,
S
chool of Medical Sciences, Universiti Sains Malaysia
,
16150, Kubang Kerian,
Kelantan Malaysia
Chee Keong Chen
Sports Science Unit
,
S
chool of Medical Sciences, Universiti Sains Malaysia
,
16150, Kubang Kerian,
Kelantan Malaysia
Asari Mohd Asnizam
Anatomy Department, School of Medical Sciences, Universiti Sains Malaysia, 16150, Kubang
Kerian, Kelantan Malaysia
ABSTRACT
Although combination of physical activity with supplementation has being investigated on its effects in maintaining and enhancing
bone health, muscular strength and immune functions, little is known about the effectiveness of combined Eurycoma longifolia Jack
supplementation and a circuit training programme on bone metabolism markers, muscular strength and power, and immune functions
in adult men. This study investigated the combined effects of 8 weeks Eurycoma longifolia Jack supplementation and circuit training
programme on bone metabolism markers, isokinetic muscular strength and power, and immune functions in adult men. Forty adult
male subjects (mean age:46.8±5.5 years), were age-matched and subsequently assigned into four groups with n=10 per group: Control
group with placebo supplementation (C), Eurycoma longifolia Jack supplementation group (ElJ), circuit training programme group
with placebo (Ex), and combined circuit training programme with Eurycoma longifolia Jack group (ElJEx). Circuit training
programme consisted of 2 circuits of exercise, with 10 different exercise stations per circuit, three times per week for a total of 8
weeks. Subjects in C and Ex group consumed placebo, subjects in ElJ and ElJEx groups consumed Eurycoma longifolia Jack, 7
days/week for 8 weeks. Before and after 8 weeks of experimental period, subjects’ anthropometry, isokinetic muscular strength and
power were measured. Blood samples were taken to determine bone metabolism markers and immune functions. ElJEx increased knee
extension peak torque at 60o.s-1, shoulder flexion average power at 300o.s-1 and shoulder extension average power at 300o.s-1.
Meanwhile, Ex alone could only increase shoulder extension average power at 300o.s-1. These results suggested that combination of
circuit training with Eurycoma longifolia Jack (ElJEx) elicited more beneficial effect on isokinetic muscular strength and power than
circuit training alone (Ex). Regarding bone metabolism markers and immune function parameters, ElJEx, Ex and ElJ did not elicit any
effect on these measured parameters after 8 weeks of experimental period. Combined Eurycoma longifolia Jack supplementation with
circuit training programme has potential to be proposed for formulating guidelines in planning exercise and nutritional promotion
programmes for the enhancement of muscular strength and power in adult men.
__________________________________________________________________________________________________________
I. INTRODUCTION
Physical activity is an important factor in attaining peak bone
mass. Bone mineral density (BMD) is a static measure of bone
mass. Changes in bone mass occur at slow rate, therefore
measurement of BMD is not enough to detect acute changes in
bone metabolism as a result of exercise training. Osteocalcin
which is produced by osteoblasts during matrix mineralization
of bone formation and several other resorption biomarkers
have enabled the evaluation of acute changes in bone turnover
in response to biological and biomechanical variables (Remes
et al., 2004). Healthy lifestyle such as carrying out regular
weight-bearing exercises during growth is one of the factors
that can contribute to bone strength. Weight bearing exercises
can increase bone density by increasing osteoblastic activity,
endogenous electrical activity and serum osteocalcin levels
(Moffat et al., 2006). Bone mineral density changes have also
been reported were associated with increments in serum levels
of osteocalcin by Casez et al. (1995). Bone is especially
adaptable to dynamic exercise, and impact-producing exercise
which produces high ground reaction force is more osteogenic
than non impact-producing exercise (Welch et al., 2008). The
development of muscle and bone during growth is influenced
by gravitational forces associated with body weight and
physical activity. Muscles become larger and stronger in
response to increased loading produce by exercise, and bones
adapt to increased loads by strong muscle contraction with
enhancement in bone mass, size, and strength. The
relationship between muscle and bone supports the concept of
COMBINED EFFECTS OF EURYCOMA LONGIFOLIA
JACK SUPPLEMENTATION AND A CIRCUIT
TRAINING PROGRAMME ON BONE METABOLISM
MARKERS, MUSCULAR STRENGTH AND POWER,
AND IMMUNE FUNCTIONS IN ADULT MEN
SPORTS SCIENCE
Keywords: bone, exercise, Eurycoma
longifolia Jack, immune function,
muscular strength, power
Research Paper
IJERSS Volume 2 | Issue 3 MAR 2015
I n t e r n a t i o n a l J o u r n a l o f E n g i n e e r i n g R e s e a r c h a n d S p o r t s S c i e n c e
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a ‘functional muscle–bone unit’, in which changes in muscle
mass and strength can affect bone in mass, size, and strength
correspondingly (Daly et al., 2004). Aging is not a factor to
limit and improve muscle strength through training, and both
older men and women have been shown to increase muscle
strength as young subjects (Ciolac and Greve, 2011).
Regarding exercise and immunity, moderate exercise was
reported has been linked to positive immune system response
and a temporary boost in the production of macrophages, the
cells that attack bacteria. It is believed that regular exercise
can lead to substantial benefits in immune system over long-
term. During moderate exercise, immune cells circulate
through the body more quickly and are efficient in killing
bacteria and viruses (Quinn, 2008). Eurycoma longifolia Jack
is from Simaroubaceae family and it is identified locally as
‘Tongkat Ali’. It is found in primary and secondary, evergreen
and mixed deciduous forests in Burma, Indochina, Thailand,
Malaysia, Sumatra, Borneo and the Philippines. It is an
essential component for the treatment of fevers, aches, sexual
insufficiency and also as health supplements. Traditional
medicinal users usually take a decoction of its roots in water
as a health tonic and antistress supplementation (Ang and
Cheang, 1999). Eurycoma longifolia Jack is reported can
increase ones testosterone level (Tambi, 2005). Thus, it is
hypothesized that Eurycoma longifolia Jack consumption may
increase muscular strength. Eurycoma longifolia Jack is also
believed can increase immune function as antimalarial, and
antibacterial. Eurycoma longifolia jack contains mainly
quassinoids, and other active compounds such as tirucallene-
type triterpenes, squalene derivatives, biphenyl-neo-lignans,
anthraquinones, canthin-6-one alkaloids, α-carboline alkaloids
and dimeric dihydrobenzofuran. Some of these compounds
were found to possess antiulcer, anthelmintic, antimalaria, and
cytotoxic activities (Teh et al., 2010). Eurycoma longifolia
Jack has been reported could enhance muscular strength and
mean arm circumference when combined with an intense
strength training programme (Hamzah and Yusuf, 2003;
Sareena and Ashril, 2002). To date, no studies have been
carried out to investigate the combined effects of a moderate
intensity circuit training and a Malaysian local product of
Eurycoma longifolia Jack supplementation on bone
metabolism markers, muscular strength and power, and
immune function in adult men, thus the present study was
proposed. The objective of the present study was to investigate
the combined effects of a 8 weeks circuit training programme
and Eurycoma longifolia Jack supplementation on bone
metabolism markers, muscular strength and power, and
immune functions in adult men. If it is found that a combined
circuit training programme and Eurycoma longifolia Jack
supplementation can elicit positive effects on enhancing bone
metabolism, muscular strength and power, and immune
functions in adult men, results obtained from this study can
then be used for formulating guidelines in planning exercise
and nutritional promotion programmes for increasing bone
health, muscular strength and power, and immune functions in
this population.
II .METHODOLOGY
Forty Malaysian male subjects with age ranging from 35 to 55
year old were recruited in this study. The inclusion criteria of
the subjects including the subjects did not exercise more than
once per week, free from any health problems, did not
consume Eurycoma longifolia Jack or any other herbs as
supplements and did not engage with any other training
programmes. The subjects were recruited among the staff of
Health Campus, Universiti Sains Malaysia. The subjects were
age matched and than assigned into four groups, with ten
subjects per group (n=10) . All subjects were given a detail
explanation about the objectives, procedures, benefits, risks
and possible discomforts experienced in this study before
giving their formal consent. This study was approved by the
Human Research and Ethical Committee of Universiti Sains
Malaysia. In the present study, the subjects were divided into
four groups, with 10 subjects per group (n = 10): Eight weeks
of sedentary with placebo control (C), eight weeks of
sedentary with Eurycoma longifolia Jack supplementation
(ElJ), eight weeks of circuit training with placebo (Ex), and
eight weeks of circuit training with Eurycoma longifolia Jack
supplementation (ElJEx) groups. Subjects in the control group
did not perform neither exercises nor taking Eurycoma
longifolia Jack supplementation. Subjects in ElJ group
consumed 400mg of ElJ for 7 days per week for a total 8
weeks duration. Subjects in group Ex were required to
perform circuit training programme for 8 weeks with 3 session
per week and one hour per session. Subjects in the combined
group, i.e. ElJEx group performed circuit training programme
for one hour per session, 3 session per week and consumed
400mg ElJ for 7 days per week for 8 weeks after breakfast for
eight weeks. The subjects in both the exercise with placebo
group (Ex) and Eurycoma longifolia Jack supplementation
with exercise group (ElJEx) were required to carry out circuit
training sessions for 3 sessions per week, one hour per session
for 8 weeks. The exercise sessions started with 10 minutes of
warm-up and ended with 5 minutes of cooling down activities.
The circuit training programme consisted of two circuits. In
each circuit, subjects performed 10 different exercises in 10
different stations (one type of exercise per station, each
subject spent 30 seconds in one particular station). The work
rest ratio was 1:2, where subjects exercised for 30 seconds for
one activity, and rested for one minute before continued with
the next activities. Resting time between circuits was five
minutes. The activities that prescribed in the circuit training
Research Paper
IJERSS Volume 2 | Issue 3 MAR 2015
I n t e r n a t i o n a l J o u r n a l o f E n g i n e e r i n g R e s e a r c h a n d S p o r t s S c i e n c e
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programme were lateral raise elastic band exercise, leg
abduction elastic band, free-weight dumbbell triceps
extension, rope skipping, free-weight dumbbell concentration
curl, sit-up, back extension, burpee, push-up and split squat.
The intensity of circuit training programme was estimated by
using heart rate monitor (polar watch, S710, US) worn by one
subjects throughout the circuit training programme. Each
subject in Eurycoma longifolia Jack group (ElJ), and
combined circuit training programme with Eurycoma
longifolia Jack supplementation group (ElJEx) consumed
Eurycoma longifolia Jack supplementation in a dose of 400mg
per day (4 capsules; 100mg per capsule) for 7 days per weeks
for a total of 8 weeks duration. The subjects were required to
consume Eurycoma longifolia Jack supplementation after
breakfast. Subjects in sedentary with placebo control group
(C) and circuit training with placebo (Ex) were required to
consume 4 placebo capsules per day, 7 days per weeks for 8
weeks. Biotropics Malaysia Berhad was the supplier of ElJ
and placebo used in this study. Anthropometry measurements
were recorded at pre and post experimental after the blood
taking on the same day. Body mass were measured using a
Body Composition Analyser (TANITA, TBF-410 Model,
Japan). The body height was measured using a body height
(SECA UK). Immediately before eight weeks of experimental
period, all the subjects were required to have blood sample
taking. Blood taking was carried out again after 8 weeks of
experimental period. Both blood taking sessions were
conducted in the morning at 8.30 a.m after a 10 hour fast of
the subjects (drinking water was permitted). The blood was
drawn by the laboratory Universiti Sains Malaysia
technologist in the Sport Science Laboratory to determine the
bone metabolism markers and immune functions parameters.
About 6 ml of blood were drawn from each subject from the
antecubital vein in seated position. Blood taking for subjects
in Ex and ElJEx were carried out 18-20 hours (8-10 am) after
performing exercise. The blood were collected into two
separate test tubes, 2 ml into an EDTA tube and another 4 ml
into a plain tube, without EDTA. In the present study, EDTA
contained whole blood was used for routine full blood count
and determination of immunophenotyping status. Blood
samples collected into EDTA tubes were brought to the
immunology laboratory and processed on the same day of
collection. An automated hematology analyser (Sysmex XS-
800i) was used for analysis of full blood count, i.e. white
blood cells (WBC), lymphocytes, neutrophil, monocytes,
eosinophil and basophil counts. Immunophenotyping analysis
of serum T lymphocytes (CD3, CD4 and CD8) and B cell
(CD19) were carried out by using a flow cytometer (BD
FACS Cantor II, Becton Dickinson, USA) with a four-colour
direct immunofluorescence reagent of a reagent kit (BD
Multitest
TM
IMK). Serum from the clotted blood in the plain
tube was used for determining serum bone metabolism status.
Serum was obtained by centrifuged the blood sample using a
centrifuge (Hettich-Rotina 46RS, Germany) for 10 minutes at
4000 rpm in 4°C temperature, before being divided into equal
portions and stored at -80°C in a freezer (ThermoForma,
Model 705, USA) for subsequent analysis. An automatic
analyzer (Hitachi Automatic Analyzer 912, Bohringer
Mannheim, Germany) with reagent kits (Randox, UK) were
used to analysed serum total calcium. Serum osteocalcin was
analysed using an enzyme immunological test kit ( N-MID®
Osteocalcin ELISA), and the concentration was determined
using a photometric microplate reader (Molecular Devices;
Versa
max
tubable microplate reader, U.S.A). Serum 1CTP is a
bone resorption marker, it was analyzed by a quantitative
enzymeimmunoassay kit Orion Diagnostica UniQ 1CTP EIA
(Finland) and the concentration was determined by a
photometric microplate reader (Molecular Devices; Versa
max
tubable microplate reader, U.S.A). An isokinetic dynamometer
(Biodex System 3 Pro, New York, USA) were used to
measure the dominant side of lower and upper limbs peak
torque (strength) and power of subjects. The subjects were
allowed to have meals before performing the tests to have
energy to do the test. Two different angular velocities were
used (60
o
.s
-1
and 300
o
.s
-1
) to measure knee and arm flexion
and extension muscular strength and power of the subjects
prior and post 8 weeks of experimental period. To begin the
test, the subject was seated while leaning against a backrest
tilted 85° from the horizontal. The subjects’ shoulder, waist,
and thigh were strap to minimize body movement and gave
stabilization position of the body during the test. Shoulder
straps were applied diagonally across the chest to prevent
excessive upper body movement. The lateral femoral
epicondyle was palpated and used as a bony landmark for
matching the axis rotation of the knee joint and the axis
rotation of the dynamometer shaft. Then, the dynamometer
was set up to measure lower limbs strength and power. The
dynamometer and the chair were rotated to 90 degrees. The
procedure was fully informed to all of the subjects before
performing this test. There were asked to do 5 repetitions for
the 60
o
.s
-1
angular velocity and 10 repetitions for the 300
o
.s
-1
angular velocity for each upper and lower limb. The subjects
were given 60 seconds to rest between each angular velocity.
All statistical analysis were performed using statistical
software in the Statistical Package for Social Sciences (SPSS)
Version 20. Repeated measure ANOVA was performed to
determine the significance of the difference between and
within groups. Statistical significance was accepted at p <0.05.
All data are expressed as means + standard deviation (SD).
Research Paper
IJERSS Volume 2 | Issue 3 MAR 2015
I n t e r n a t i o n a l J o u r n a l o f E n g i n e e r i n g R e s e a r c h a n d S p o r t s S c i e n c e
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III. ANALYSIS OF DATA AND RESULTS
Physical Characteristics of the Subjects
Forty healthy sedentary adult men with mean age 46.80 ± 5.52
years old completed the present study. There were no
significant differences between groups in age and body height
and body mass at the beginning of the experimental period
(Table 1). At the end of the study, no statistical significant
differences in body mass were observed among all groups.
Table 1: Means age, body height and body mass of the
subjects
Bone Metabolism Markers
Results of mean total calcium concentrations of all the groups
are presented in Table 2. In the pre and post tests, there were
no significant differences in ElJ, Ex and ElJEx groups
compared to respective C group respectively. After 8 weeks of
experimental period, no significant differences were observed
in serum total calcium concentrations between pre and post
tests in all the groups. Table 2 illustrated results of bone
formation marker of serum osteocalcin concentrations in all
the groups at pre and post tests. In pre test, there was
significant difference (p<0.05) in ElJ group compared to C
group respectively. In post test, there was significant
differences (p<0.05) in serum osteocalcin between Ex group
and ElJ group. After 8 weeks of experimental period, there
were no significant differences in serum osteocalcin in post
test compared to pre test in all the groups. Mean serum C-
terminal telopeptide of type 1 procollagen (1CTP)
concentrations of all groups can be seen in Table 2. In pre
test, there were no significant differences in all the ElJ, Ex,
and ElJEx groups compared to respective C group
respectively. However, in post test there was significant
difference (p<0.05) between ElJ group and C group.
Significant difference (p<0.05) was also observed between
ElJEx group and ElJ group. After 8 weeks of study period,
there were no significant differences in serum 1CTP
concentrations between pre and post tests in all groups.
Table 2. Mean serum total calcium, serum osteocalcin, serum
1CTP concentrations (Mean±SD)
#
significantly different from respective control group
(p<0.05)
&
significantly different from respective ElJ group (p<0.05)
Muscular Strength (Peak Torque) and Power
Figure 1(a) exhibited the results of knee flexion isokinetic
peak torque at 60
o
.s
-1
in all the groups at pre and post tests. In
pre test, there were significant differences (p<0.05) in ElJ, Ex
and ElJEx groups compared to C group respectively. In post
test, there were significant differences (p<0.05) in Ex and
ElJEx groups compared to C group respectively. After 8
weeks, there were statistically significant greater values of
knee flexion peak torque at 60
o
.s
-1
in C and Ex groups in post
test compared to pre test. Results of knee extension isokinetic
peak torque at 60
o
.s
-1
in all the groups at pre and post tests are
shown in Figure 1(b). In pre and post tests, there were no
significant differences in ElJ, Ex, and ElJEx groups compared
to C group respectively. After 8 weeks of experimental period,
statistically greater value of isokinetic knee extension peak
torque at 60
o
.s
-1
was found in ElJEx group compared to pre
test. Figure 1(c) showed the result of knee flexion average
power at 300
o
.s
-1
in all the groups at pre and post tests. In pre
test, there were no significant differences in all the ElJ, Ex and
ElJEx groups compared to C group respectively. In post test,
there were significant differences (p<0.05) in Ex and ElJEx
groups compared to C and ElJ groups respectively. After 8
weeks of experimental period, no statistically significant
differences in knee flexion average power at 300
o
.s
-1
was
found between pre and post tests in all the groups. Study
findings of knee extension average power at 300
o
.s
-1
in all the
groups at pre and post tests are shown in Figure 1(d). In pre
and post tests, there were significant differences (p<0.05) in
Ex group compared to C and ElJ groups respectively. After 8
weeks of experimental period, there were statistically
significant greater values of knee extension average power at
Research Paper
IJERSS Volume 2 | Issue 3 MAR 2015
I n t e r n a t i o n a l J o u r n a l o f E n g i n e e r i n g R e s e a r c h a n d S p o r t s S c i e n c e
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300
o
.s
-1
in C, Ex and ElJEx groups compared to pre test.
Figure 1: Knee flexion (a) and extension (b) peak torque at 60
o
.s
-1
and knee flexion (c) and extension (d) average power at 300
o
.s
-1
at
pre and post tests (Mean±SD)
# significantly different from respective control group
(p<0.05)
& significantly different from respective ElJ group (p<0.05)
* significantly different from pre test (p<0.05)
Figure 2: Shoulder flexion (a) and extension (b) peak torque at 60
o
.s
-1
and shoulder flexion (c) and extension (d) average power at 300
o
.s
-1
at pre and post tests (Mean±SD)
# significantly different from respective control group
(p<0.05)
* significantly different from pre test (p<0.05)
Figure 2(a) illustrated result of shoulder flexion peak torque at
60
o
.s
-1
in all the groups at pre and post tests. In pre test, there
were no significant differences in ElJ, Ex and ElJEx groups
compared to respective C group respectively. In post test,
there were significant difference (p<0.05) between Ex and
ElJEx groups compared to C group respectively. After 8
weeks, no statistically significant differences in shoulder
flexion peak torque at 60
o
.s
-1
were observed between pre and
post test in all the groups. Result of shoulder extension peak
torque at 60
o
.s
-1
in all the groups at pre and post tests are
shown in Figure 2(b). In pre and post test, there were
significant differences (p<0.05) in ElJ groups compared to C
group. After 8 weeks, no statistically significant differences
were observed between pre and post test in isokinetic shoulder
extension peak torque at 60
o
.s
-1
in all the groups. Findings of
shoulder flexion peak torque at 300
o
.s
-1
in all the groups at pre
and post tests are shown in Figure 2(c). In pre test, there were
no significant differences in ElJ, Ex and ElJEx groups
compared to C group respectively. In post test, there was
significant difference (p<0.05) between ElJ group and C
group. After 8 weeks, there was statistically significant greater
value of shoulder flexion average power at 300
o
.s
-1
in ElJEx
compared to pre test. Figure 2(d) exhibited result of shoulder
extension average power at 300
o
.s
-1
in all the groups at pre and
post tests. In pre test, there was significant difference (p<0.05)
between ElJEx and C group. In post test, there were no
significant differences in ElJ, Ex and ElJEx groups compared
to C group respectively. After 8 weeks, there were statistically
significant greater values of shoulder extension average power
at 300
o
.s
-1
in Ex and ElJEx groups compared to pre test.
Immune Functions
Study findings of white blood cells (WBC), neutrophils and
lymphocytes count in all the groups at pre and post tests are
presented in Table 3. In pre test, WBC count concentrations
were significantly different (p<0.05) in Ex and ElJEx groups
compared to C group respectively. There was also significant
difference (p<0.05) in ElJEx group compared to ElJ group. In
post test, there were no significant differences in ElJ, Ex and
ElJEx groups compared to C group respectively. After 8
weeks of study period, there were no statistically significant
differences in white blood cells in all the groups in post test
compared to pre test. In pre test, neutrophil counts were
significant different (p<0.05) in ElJEx group compared to C
and ElJ group respectively. In post test, there were no
significant differences in ElJ, Ex and ElJEx groups compared
to C group respectively. After 8 weeks of study period, there
were no statistically significant differences in neutrophil count
in all the groups in post test compared to pre test. Lymphocyte
count was significant different (p<0.05) in Ex group compared
to C group respectively in pre test. In post test, there were no
significant differences in ElJ, Ex and ElJEx groups compared
to C group respectively. After 8 weeks of study period, there
were no statistically significant differences in lymphocyte
count in all the groups in post test compared to pre test.
Research Paper
IJERSS Volume 2 | Issue 3 MAR 2015
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Table 3: Mean white blood cells (WBC), neutrophil and total
lymphocyte counts
# significantly different from respective control group
(p<0.05)
& significantly different from respective ElJ group (p<0.05)
Results of monocyte, eosinophil and basophil counts in all the
groups at pre and post tests are presented in Table 4. In pre
test, monocyte count was significant different (p<0.05) in Ex
group compared to ElJ group. In post test, there were no
significant differences in ElJ, Ex and ElJEx groups compared
to C group respectively. After 8 weeks, there were no
statistically significant differences in monocyte count in all the
groups in post test compared to pre test. In pre and post test,
eosinophil counts were significant different (p<0.05) in ElJEx
group compared to C, ElJ and Ex groups respectively.
However, after 8 weeks of experimental period, there were no
statistically significant differences observed in eosinophil
count in all the groups in post test compared to pre test. In pre
test, basophil count was significant different (p<0.05) in Ex
group compared to C and ElJ group respectively. In post test,
there were no significant differences in ElJ, Ex and ElJEx
groups compared to C group respectively. After 8 weeks of
experimental period, there was significant lower value of
basophil in ElJ group in post test compared to pre test.
Table 4: Mean monocyte, eosinophil and basophil counts
# significantly different from respective control group
(p<0.05)
& significantly different from respective ElJ group (p<0.05)
+ significantly different from respective Ex group (p<0.05)
* significantly different from pre test (p<0.05)
Study results of total T cells (CD3) count in all the groups at
pre and post tests are presented in Table 5. In pre and post
tests, there was significant difference in ElJEx group
compared to C group respectively. However, after 8 weeks of
experimental period, there were no statistically significant
differences in total T cells (CD3) in all the groups in post test
compared to pre test. Table 5 illustrated results of T
helper/inducer (CD4) cell count in all the groups at pre and
post tests. In pre and post tests, there were no significant
differences in ElJ, Ex and ElJEx groups compared to C group
respectively. After 8 weeks, there were no statistically
significant differences in T helper/inducer (CD4) cell count in
all the groups in post test compared to pre test. Regarding T
cytotoxic/ suppressor (CD8) count in all the groups at pre and
post tests as shown in Table 5, it was found that there were
no significant differences in ElJ, Ex and ElJEx groups
compared to C group respectively in pre test. However, in post
test, there was significant difference (p<0.05) between ElJEx
group and C group. After 8 weeks, there were no statistically
significant differences in T cytotoxic/suppressor (CD8) cell
count in all the groups in post test compared to pre test. Result
of total B cells (CD19) counts in all of the groups at pre and
post tests can be seen in Table 5. In pre and post tests, there
were no significant differences in ElJ, Ex and ElJEx groups
compared to C group respectively. After 8 weeks, there were
no statistically significant differences in total B cells (CD19)
count in all the groups in post test compared to pre test.
Table 5: Mean total T cells (CD3), T helper/inducer (CD4), T
cytotoxic /suppressor (CD8), Total B cells(CD19) cell counts
(Mean±SD)
# significantly different from respective control group
(p<0.05)
IV. FINDINGS & CONCLUSIONS
Understanding the reaction of bone to physical exercise is
important for developing strategies to achieve higher bone
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mass and prevent bone loss. One of the main objectives of the
study was to investigate the effects of combined circuit
training exercise with Eurycoma longifolia Jack
supplementation on bone metabolism markers. The present
study found that serum osteocalcin, a bone formation marker
was not affected by Eurycoma longifolia Jack
supplementation, exercise nor combination of both. Similarly,
there was no significant main effect of time on serum 1CTP, a
bone resorption marker between pre and post tests for all the
groups. In serum total calcium, the present study also found
that there were no significant differences between pre and post
tests for all the groups. In a previous study done by Maimoun
et al. (2005), it was shown that bone metabolism markers such
as serum osteocalcin and 1CTP did not change after exercise
in active elderly subjects. The authors mentioned that bone
tissue of elderly population may have become less responsive
to exercise. This was supported by Wallace et al. (2000)
whomentioned that age is probably a negative determinant of
the bone marker response to exercise. Similarly, the present
study findings with no significant changes in bone metabolism
markers in men with mean age of 47 years may imply that
bones are less responsive to exercise with middle aged men.
Bassey and Ramsdale (1995), reported that there was no
significant increase in bone mineral density of post
menopausal women who performed 12 months of weight-
bearing exercise. The authors speculated that postmenopausal
women are with low blood estrogen levels, and this could have
reduced the potential osteogenic response. In the present
study, the mean age of the subjects was 47 years old, and their
hormone levels could be one of the factors in influencing the
present study results on bone metabolism markers. Changes in
bone metabolism with exercise was also reported by Remes et
al. (2004), who found that serum osteocalcin decreased in
middle aged men after 4 years of performing low to moderate
exercise, and the authors concluded that bone turnover in men
may be influenced by circulating sex hormone levels. The
discrepancy between the finding of this previous study and the
present study could be attributed to differences in the duration
and the type of exercise prescribed. With regards to effects of
sex hormones on bone metabolism markers, Falahati et al.
(2000) found that bone resorption markers increased while
serum osteocalcin decreased significantly in the absence of
testosterone and estradiol in elderly men with mean age of 68
years. Eurycoma longifolia Jack was reported to have
increased one’s testosterone level (Tambi, 2005), but we
found that Eurycoma longifolia Jack supplementation alone
did not elicit any positive effects on bone metabolism markers
in our subjects in the present study. Hence, future studies with
longer experimental period and with measurement of blood
testosterone are warranted to confirm the beneficial effects of
this supplementation on bone. Bellometti et al. (2002) found
that physical exercise could not increase alkaline phosphatase,
a type of bone formation marker in osteoarthritic aging
women. However, Lester et al. (2009) found that in inactive
young women, serum bone alkaline phosphatase concentration
increased by 15.8% and serum osteocalcin increased by 8.5%
in response to eight weeks of combined aerobic and resistance
exercises, even though there were no changes in bone
resorption markers (1CTP) after eight weeks of the combined
exercise programme. This finding may reflect that bone
formation can occur at a greater rate in younger participants
compared to an older cohort with exercise training and bone
metabolism in a person can be reduced with aging. Regarding
combined effects of exercise and nutritional supplementation
on bone formation marker in humans, it was found by the
present research team that bone formation marker which is
serum alkaline phosphatase, significantly increased in young
female subjects after 6 weeks of consuming honey combined
with 6 weeks of aerobic dance exercise (Ooi et al., 2011).
Similarly, Lau and Ooi (2014) also reported that combined
circuit training programme and chocolate malt drink
supplementation elicited significant effect on reducing bone
resorption marker i.e. 1CTP in young males. The lack of
significant changes in bone metabolism markers in the
combined Eurycoma longifolia Jack and circuit training group
compared to the findings of our previous studies mentioned
above may be due to the difference in of the type of
supplementation prescribed and the age of the subjects
particularly. With regards to effects of exercise on muscular
strength, Dias et al (2010) found that muscular strength
increased in untrained young men after 8 weeks of exercise.
Kida et al. (2008) also found that muscular strength increased
in men with mean age 60 years after performing 3 months of
treadmill running exercise. In their study, subjects’ peak
torque for knee extension and flexion at 60
o
.s
-1
increased after
3 months of exercise. Similar increases in muscular strength
reflected by peak torque were observed with ElJEx. In Weng
et al. (2009), male and female knee osteoarthritic patients with
mean age 64 years old showed significant improvement in
muscular strength reflected by 60
o
.s
-1
angular velocity peak
torques, after following an isokinetic exercise programme at
three times per week for 8 weeks. In the present study,
combined group of ElJEx showed more beneficial effects than
Ex alone in increasing muscular strength, and this could be
due to Eurycoma longifolia Jack which was reported to be
able to increase testosterone level (Tambi, 2005) and
subsequently increase muscular strength, when combined with
exercise, it can elicit better positive effect in enhancing muscle
strength than exercise alone. In muscular power, discernable
significant increase in power of shoulder flexion at 300
o
.s
-1
was observed in ElJEx group. Additionally, discernable
significant increase in power of shoulder extension at 300
o
.s
-1
was observed in both ElJEx and Ex groups. In a previous
study by Treiber et al. (1998), it was found that resistance
training programme by using theraband elastic tubing and
lightweight dumbbells could increase shoulder strength and
power after following 4 weeks of exercise programme. Similar
to this previous study, the prescribed circuit training in the
present study also included dumbbells and elastic bands and
significant increases in power of shoulder extension and
shoulder flexion at 300
o
.s
-1
were observed
in both Ex and
ElJEx groups. These findings imply that these equipments
were suitable for enhancing muscular power. Beneka et al.
(2005) mentioned that high intensity training protocol can
cause the most impressive improvement of knee extensors’
isokinetic performance as compared to low intensity or
Research Paper
IJERSS Volume 2 | Issue 3 MAR 2015
I n t e r n a t i o n a l J o u r n a l o f E n g i n e e r i n g R e s e a r c h a n d S p o r t s S c i e n c e
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8
moderate intensity at all testing velocities in elderly men and
women. However, moderate and low intensity training can
cause a significant improvement in strength as well but at a
lesser extent. In the present study, each of the subject in the
group of Ex and ElJEx carried out the circuit training
exercises, with exercise heart rate ranging from 110 to 120
beats.min
-1
i.e. approximately 60%-70% of the prescribed
maximum heart rate of the subjects. Thus, the present findings
have confirmed that moderate intensity circuit training could
also elicit significant beneficial effects on muscular
performance in middle aged men. In the present study, it was
shown that there were no significant changes in white blood
cells, neutrophil, lymphocyte, eosinophil, monocyte, basophil,
CD3 T cells, CD4 T cells, CD8 T cells and CD19 B cells in
the groups of ElJ, Ex and ElJEx in post test compared to pre
test. These findings imply that ElJ alone, Ex alone and
combined ElJEx could not improve or increase immune
functions in adult men. The present findings with exercise
alone on immune function parameters reflect that the immune
status of subjects did not change with the prescribed exercise
programme. Vider et al. (2001) found that there were increases
in leukocytes, lymphocytes subsets, neutrophils and B
lymphocytes immediately after exercise. Nevertheless, the
proportions declined after 30 minute of exercise during
recovery phase. In the present study, the blood withdrawal of
the subjects at post test after 8 weeks of experimental period
were carried out 18 to 20 hours after subjects performed
circuit training. The differences in the blood taking time may
have caused the differences in the present study results
compared to study result of Vider et al (2001). Nieman et al.
(1993) found that a 3 month moderate aerobic exercise
programme failed to increase NK cell cytotoxicity, T
lymphocyte mitogenesis, NK cell or T cell subsets in
sedentary women. Woods et al. (1999) also found that there
were no intervention-induced changes in total white blood
cell, neutrophil, lymphocyte, monocyte, eosinophil, or
basophil blood counts in the elderly after 6 months of
moderate aerobic exercise training. Furthermore, the
percentage and number of CD3, CD4 and CD8 T cells in the
blood remained unchanged. Similarly, this present study
findings also did not show any positive effects of 8 weeks of
exercise on immune function parameters. It was speculated
that the duration of exercise of 8 weeks in the present study
was not long enough to elicit positive effects on immune
function parameters Starkie et al. (2005) reported that there
were increases in circulating neutrophil, monocytes and
lymphocyte concentrations in the subjects immediately after a
cycling exercise. However, after 2 hours of post exercise, only
monocyte and neutrophil remained elevated whereas
lymphocyte concentration returned to pre-exercise levels. The
subjects in this previous study were young endurance trained
men with mean age of 26 years old. Hence, the age of the
subjects maybe one of the contributing factors that caused
equivocal findings of this previous study with the present
study, where low stimulation of immune function may occur
in older subjects compared to younger subjects. Aging appears
to have an effect on the ability of the lymphatic system to
remove fluid from tissues, absorb nutrients from the digestive
tract, or remove defective red blood cells from the blood.
Aging also seems to have a direct effect on the ability of B
cells to respond to antigens, and the number of circulating B
cells remaining stable in most individuals (Seeley et al., 2006).
With age, thymic tissue is replaced with adipose tissue and the
ability to produce new and mature T cells in the thymus is
eventually lost. The number of T cells remaining stable in
most individuals is due to the replication of T cells in
secondary lymphatic tissues. In many individuals, however,
there is a decreased ability of helper T cells to proliferate in
response to antigens. Thus, antigen exposure produces fewer
helper T cells which results in less stimulation of B cells and
effector T cells. Consequently, both antibody mediated
immunity and cell-mediated immunity responses to antigens
decrease (Seeley et al, 2006). The above explanation could be
the reasons for the insignificant findings on immune functions
in the middle aged subjects with mean age of 47 years in the
present study. In a previous study carried out by the present
research team (Muhamad et al., 2009), it was found that
supplementation of Eurycoma longifolia Jack did not cause
any changes in white blood cells, monocytes, basophils,
neutrophils, eosinophils, platelets, CD3 T cells, CD4 T cells,
CD8 T cells and CD19 B cells after a bout of running in the
heat in recreational athletes. The subjects consumed 150mg of
ElJ for 7 days before the running trials. In the present study,
there were no changes in blood immune function parameters
after consuming 400 mg of Eurycoma longifolia Jack
combined with exercise for 8 weeks. In summary, the present
study found that combined circuit training programme and
Eurycoma longifolia Jack supplementation elicited more
beneficial effects on isokinetic muscular strength and power
compared to circuit training programme alone or Eurycoma
longifolia Jack alone in adult men. In bone metabolism
markers, ElJ, Ex and ElJEx groups did not elicit positive
effect on serum total calcium, osteocalcin, and C-terminal
telopeptide of type 1 procollagen (1CTP) concentrations. With
regards to muscular strength and average power, combined
circuit training with Eurycoma longifolia Jack (ElJEx) elicited
beneficial effects on isokinetic muscular strength and power,
and circuit training alone (Ex) could only increase power.
Combined circuit training with Eurycoma longifolia Jack
(ElJEx) elicited more beneficial effect on isokinetic muscular
strength and power than circuit training alone (Ex). In immune
functions, the combination of circuit training and Eurycoma
longifolia Jack (ElJEx), circuit training alone (Ex) and
Eurycoma longifolia Jack (ElJ) did not elicit positive effect on
immune function parameters. In conclusion, supplementation
of Eurycoma longifolia Jack (400mg/day) with circuit training
programme (3 days/week) can be recommended for increasing
muscular strength and power in adult men. Additionally, this
combination has potential to be proposed for formulating
guidelines in planning exercise and nutritional promotion
programmes for the enhancement of muscular strength and
power in adult men.
Research Paper
IJERSS Volume 2 | Issue 3 MAR 2015
I n t e r n a t i o n a l J o u r n a l o f E n g i n e e r i n g R e s e a r c h a n d S p o r t s S c i e n c e
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ACKNOWLEDGEMENT
We wish to extend our sincere gratitude to all the subjects who
have participated in this study. We also want to express our
appreciation to Mdm. Jamaayah bt. Meor Osman, Mdm.
Norlida bt Azalan, Mdm. Hafizah bt. Hamzah and Mdm. Nor
Aini bt. Sudin from Sports Science Unit, Universiti Sains
Malaysia for their technical assistance. Special thanks to Mr.
Jamaruddin Mat Asan from Immunology Department for his
assistance. We would like to thank Biotropics Malaysia
Berhad, Kuala Lumpur, Malaysia for partially financing this
study.
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