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Medicina Sportiva (2013), vol.

IX, no 3, 2198-2202
Romanian Sports Medicine Society


Beneficial effect of aerobic exercise and green tea on serum antioxidants

Amani Sandyani S1, Erfani Karimzadeh Toosi A2, Sariri R3, Razmi N1
Science and Research Branch, Islamic Azad University, Fars, Iran
Gastrointestinal and Liver Disease Research Center, Guilan University of Medical Sciences,
Rasht, Iran 3Department of Biochemistry, University of Guilan, Rasht, Iran

Abstract. Introduction. Production of free radicals is a natural process during various metabolic events. The process
may be altered due to various internal and external factors. Increased antioxidant capacity is one of the responses of
human body against attack of free radicals. Many factors may affect the antioxidant capacity with various potential
values in health and disease. The aim of this study was assessment of antioxidant potential in plasma after aerobic
exercise, consumption of green tea and combination of both. Material and Method. Three groups of 35 healthy women
(mean age 30.11.5 years) entered the study. The first group was composed of those attending an aerobic training
course, the second group agreed to drink 3 mugs of green tea a day and the last group agreed to do both tests. Their
blood samples were collected three times during the study, i.e. beginning, after two weeks and at the end. Results. A
significant increase in catalase activity and uric acid concentration together with decreased concentration of
malondialdehyde was observed. Conclusions. Enhanced activity of antioxidants in plasma is the beneficial effect of
aerobic exercise, green tea and, most interesting, synergism of both. Since production of malondialdehyde is marker of
cell damage, its reduced concentration after the period of treatment is indicative of increased natural resistance of the
body to free radical attack.
Key words: serum antioxidants, malondialdehyde, aerobic exercise, green tea, health.

Free radicals are reactive oxygen or nitrogen the important aims of this research was to
species, having unpaired electrons. Within a determining whether MDA serum levels are
living organism, free radicals can produce harmful associated with health improving habits including
effects due to their tendency to react with proteins regular exercise and supplementation with natural
and DNA by absorbing their electrons. Production antioxidants. We assumed that MDA serum levels
of free radicals is a natural process during various may be used as biomarkers to predict health
metabolic events. The process may be altered due status.
to various internal and external factors. Increased The antioxidant system in human serum protects
antioxidant capacity is one of the responses of body form attack of free radicals. The system
human body against attack of free radicals. contains enzymatic and non-enzymatic
Malondialdehyde (MDA) is a low molecular mechanisms, both of which may change in
weight aldehyde that is formed due to the attack response of external and internal factors. The
of oxidative species on polyunsaturated fatty acids most important antioxidant enzymes are catalase,
present in the phospholipid structure of cell peroxidase and superoxide dismutase. On the
membrane. In the case of an imbalance between other hand, non-enzymatic antioxidants are
oxidants and antioxidants sever oxidative damage smaller molecules such as ascorbic acid, a water
may be resulted. This condition includes oxidative soluble vitamin, and uric acid, the end product of
modification of cellular macromolecules, purine catabolism.
induction of cell death by apoptosis or necrosis It has been recommended that to maintain cardio
and structural tissue damage. Measurement of any respiratory fitness a program of aerobic type
alternation in the level of MDA, therefore, can be exercise for 20-60 min, three to five times per
considered a valuable screening tool as a week at intensities of 50-80% VO2max is necessary
biomarker of oxidative damage (1). Thus, one of for healthy adults (2).
Beneficial effect of aerobic exercise and green tea on serum antioxidants
Amani Sandyani S & all


The beneficial effect of aerobic exercise is mostly (Figure 1). Most of these compounds act as
related to increase in the antioxidant capacity of effective scavengers of free radicals (4, 5).
human body. Therefore, absorption of the reactive Polyphenols present in green tea are considered to
oxygen (nitrogen) species is accelerated leading to be responsible for the anticarcinogenic (6), control
reduced attack to important biological molecules of harmful irradiation (7), atherosclerosis (8) and
(3). protection against cardiovascular diseases effects
On the other hand, it is also known that green tea of green tea (9, 10).
is a powerful antioxidant due to the presence of The aim of this research was investigating the
chemical compounds such as epigallocatechin alternations in antioxidant power of plasma due to
gallate (EGCG), epicatechin gallate (ECG), aerobic exercise, consumption of green tea and
epigallocatechin (EGC) and epicatechin (EC) combined effect of both.

Figure 1. Chemical structures of some important tea catechins (4)

Material and method

Phosphoric acid, thiobarbituric acid and n-butanol Subjects and collection of serum samples. Three
were purchased from Merck a local groups of 35 healthy women (mean age 30.11.5
representative. years, entered the study (Table I). Group I were
The uric acid kit was also obtained from local instructed to drink 3 mugs of green tea a day.
representatives in Iran. The whole necessary Group II were told to register in an aerobic
solutions and buffers were prepared freshly within training salon and perform 45 minutes a day
our research laboratory in double distilled water. (similar aerobic exercise under guidance of the
All general chemicals, solvents and reagents were same instructor) and group III drank the 3 mugs
obtained with highest purity available. green tea a day and registered in the aerobic
Catalase activity Assay Kit (ECAT-100), was training salon similar to group II. Their blood
purchased from a local importer in Iran. The kit samples were collected after 8 hours fasting the
was designed by EnzyChromTM and day before beginning of the study, two and four
recommended for direct assay of catalase in weeks after the beginning of study. The blood
various biological fluids including serum, saliva samples were centrifuged and their serum kept
and urine. frozen until use.

Table I. Main demographic characteristic of women subjects entered the study

Age (years) Mass (kg) Height (m) Body fat %
Mean: 23.22 62.15 1.62 19.75
SD: 2.28 7.12 0.12 3.84
Range: 35-40 52.9-72.8 1.51-1.71 18.2-21.1

Determination of uric acid. The concentration of uricase on uric acid to form allantoin and
uric acid in serum samples was determined using hydrogen peroxide (11). Production of hydrogen
a method based on the enzymatic reaction of peroxide was coupled with catalytic oxidation of

Medicina Sportiva
Beneficial effect of aerobic exercise and green tea on serum antioxidants
Amani Sandyani S & all

p-hydroxybenzoate and 4-aminoantipyrine Statistical analysis. All determinations were
oxidation in the presence of peroxidase. The pink carried out in three triplicate and data were
chromophore thus formed was then detected at subjected to analysis of variance. Analysis of
505 nm. variance was performed using the ANOVA
Catalase assay. Catalase activity was measured procedure. Statistical analyses were performed
using EnzyChromTM Catalase Assay Kit as according to the MSTATC software. Significant
decribed earlier (12). The assay method designed differences between means were determined by
so that the degradation of H2O2 by catalase was Duncans multiple range tests. P values less than
followed by a redox dye. The change in color 0.05 were considered statistically significant.
intensity at 570nm, proportional to the catalase
activity in the sample, was measured Results and Discussion
spectrophotometrically. Concentration of serum uric acid. Figure 2 shows
One unit of catalase activity was the amount of the concentration of uric acid in all three groups
catalase that decomposes 1 mole of H2O2 per min before, during and after treatment. It can be seen
at pH 7.0 and room temperature. The activity was in this figure that the same pattern of increase is
stated as (U/ml). also observed in uric acid concentration after two
Serum MDA analysis. The level of MDA in serum and four weeks in all cases, indicating an
(mmol/L) was measured using thiobarbituric acid- enhanced antioxidant power of serum due to both
reactive substance (TBARS) method (13). The exercise and green tea drinking. However, the
resulting pink complex of samples was extracted variation was statistically significant except when
in n-butanol. regularly drinking green tea was combined with a
The samples were placed in a 96-well plate and proper training of aerobic exercise. This is an
read at 535 nm in a microplate spectrophotometer interesting result as uric acid moderately soluble
reader (Benchmark Plus, Bio-Rad, Hercules, CA, in biological environment and its high
USA). concentration in serum could cause its deposition,
The detection limit for TBARS assay was 0.079 especially at joints, a condition known as gout.
nmol/ml; the intra-and inter-assay CV were 1.82% However, its higher concentrations in Group III
and 4.01%, respectively. after four weeks exercise and drinking green tea
Serum samples were kept frozen and examined all could provide a higher antioxidant activity against
at the same time, in order to avoid the possible attack of free radicals and diminish the rate of
dispersion of MDA serum level results. apoptosis

Figure 2. Variation of uric acid (mg/dl) in the serum of volunteers.

The data used to obtain the graph are mean values from 35 subjects in each group.
*P values less than 0.05 were considered statistically significant

Catalase activity. Table II shows the activity of ed in saliva according to our literature survey, the
catalase in saliva samples of both groups. It can activity of this enzyme is also important in the
be seen that the activity of this enzyme is also defense system of saliva. Therefore, its reduction
reduced in subjects taking only vegetables. could be compensated by altering other styles of
Although catalase has not been extensively studi- life, i.e. increasing exercise (14).

Medicina Sportiva
Beneficial effect of aerobic exercise and green tea on serum antioxidants
Amani Sandyani S & all

Table II. Mean SD serum catalase activity (U/l) of subjects performed the instructions for one month
Sample Group I Group II Group III P values*
Beginning of the study 3.22 0.24 3.47 0.18 3.17 0.31 0.03
Two weeks after start 3.59 0.25 4.07 0.31 10.98 0.28 0.04
Four weeks after start 12.88 0.43 5.43 0.22 5.03 0.21 0.05
Values presented as Mean SD. P values less than 0.05 were considered statistically significant

Concentration of malondialdehyde. The results bination of exercise and green tea drink has
obtained from serum MDA assay are presented in caused a balance between oxidant/antioxidant
Figure 3. It was found that serum MDA level was state in serum. It has been found that the
lower in all subjects in three groups after two imbalance of oxidant/antioxidant ration could lead
weeks from the start of treatment. This decrease in to an increase of free radicals contributing to
MDA concentration was continued up to four cellular dysfunction and accelerated aging (18).
weeks, the whole duration of study. According to However, a more extended study using a larger
our findings, combination of an aerobic exercise sample size and measurement of other indicators
and a natural antioxidant drink could act as a of oxidation such as activity of peroxidase and
modulator of antioxidant/oxidant state in healthy superoxide dismutase as well as non-enzymatic
women. This result is supported by other studies antioxidant such as ascorbic acid are still going in
using melanin and other synthetic drugs (15). It our laboratory. By measuring all such compounds
has also been reported that the concentration of of oxidant and antioxidant states we would then
malondialdehyde (MDA) in serum of critically ill be able to better evaluate this type of treatment.
septic patients (16) and those suffered from In pharmacological world, new classes of
diabetes (1) is increased deramatically. It is worth synthetic drugs acting as modulators of
reminding that MDA is a marker for accelerated antioxidant/oxidant state are introduced for
cell death due to apoptosis or necrosis as well as treatment of diseases caused due to oxidative
structural tissue damage. Therefore, combined damage such as neurodegenerative diseases and
effect of a regular aerobic exercise and drinking severe sepsis (16). Therefore, synergism of
green tea is highly beneficial in reducing the therapies such as exercise and use of natural
oxidative damage caused by imbalance between antioxidant supplements develops the hope that, in
oxidants and antioxidants. future, it could replace the use of many synthetic
According to the data obtained in this study, com- chemicals.

Figure 3. Concentration of serum MDA (mmol/L) in three volunteered groups.

The data used to obtain the graph are mean values from 35 subjects in each group
*P values less than 0.05 were considered statistically significant

The results obtained from this study demonstrated concentration and catalase activity. The
that a considerable variation in antioxidant power antioxidant activity of serum increased
of serum samples determined by uric acid significantly after one month of treatment with

Medicina Sportiva
Beneficial effect of aerobic exercise and green tea on serum antioxidants
Amani Sandyani S & all

aerobic training and drinking green tea. On the 8. Cai Y, Kurita-Ochiai T, Hashizume T, Yamamoto
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Franchini, Emmanuel J. Favaloro, Giovanni
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Acknowledgements. The authors express their 12. Sariri R , Damirchi A, Nazari Y (2013). Salivary
sincere thanks to University of Guilan for antioxidant variations in athletes after intense
financial support. exercise. Medicina Sportiva (Romanian Sports
Medicine Society); IX(1): 2043- 2050.
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