Aerobic and anaerobic training effects on the antioxidant enzymes of the blood


Selamoglu, S.; Turgay, F.; Kayatekin, B.M.; Gönenc, S.; Yslegen, C.

Acta Physiologica Hungarica 87(3): 267-273

2001


The purpose of the present study was to investigate the effects of aerobic and anaerobic training on serum lipid peroxidation levels and on antioxidant enzyme activities. Long distance runners for aerobic training group, and wrestlers for anaerobic training group were chosen. Non-sporting men were used as control group. When the aerobic power was compared; indirect VO2max of long-distance runners were found higher than wrestlers and control group (p<0.001, p<0.001). When lipid peroxidation levels were compared; levels of the thiobarbituric acid reactive substances (TBARS) of long distance runners were found to be lower than those in the control group (p<0.05), but similar to those found in wrestlers. Comparison of antioxidant enzyme activities in erythrocytes show that there were no significant difference among the groups in superoxide dismutase enzyme activities, but glutathione peroxidase (GPx) activity of long distance runners was higher than that measured in wrestlers (p<0.05). These results suggest that aerobic training increased in erythrocytes GPx activity with a subsequent decrease in plasma TBARS levels but anaerobic training had no effect on this process.

Acta
Physiologica
Hungarica,
Volume
87
(3),
pp.
267-273
(2000)
Aerobic
and
anaerobic
training
effects
on
the
antioxidant
enzymes
of
the
blood
Semih
Selamoglul,
Faruk
Turgayl,
Berkant
Muammer
Kayatekin
2
,
Sevil
Gonenc
2
,
cetin
Yslegen
3
1
Izmir
Sports
Health
Center
2
Department
of
Physiology,
Dokuz
Eyliil
University
Medical
School
3
Department
of
Sports
Medicine,
Ege
University
Medical
School,
Izmir,
Turkey
Received:
September
4,
2000
Accepted:
December
18,
2000
The
purpose
of
the
present
study
was
to
investigate
the
effects
of
aerobic
and
anaerobic
training
on
serum
lipid
peroxidation
levels
and
on
antioxidant
enzyme
activities.
Long
distance
runners
for
aerobic
training
group,
and
wrestlers
for
anaerobic
training
group
were
chosen.
Non-sporting
men
were
used
as
control
group.
When
the
aerobic
power
was
compared;
indirect
VO
2
max
of
long-distance
runners
were
found
higher
than
wrestlers
and
control
group
(p.)
0.001,
p.)
0.001).
When
lipid
peroxidation
levels
were
compared;
levels
of
the
thiobarbituric
acid
reactive
substances
(TBARS)
of
long
distance
runners
were
found
to
be
lower
than
those
in
the
control
group
(p.)
0.05),
but
similar
to
those
found
in
wrestlers.
Comparison
of
antioxidant
enzyme
activities
in
erythrocytes
show
that
there
were
no
significant
difference
among
the
groups
in
superoxide
dismutase
enzyme
activities,
but
glutathione
peroxidase
(GPx)
activity
of
long
distance
runners
was
higher
than
that
measured
in
wrestlers
(p.)
0.05).
These
results
suggest
that
aerobic
training
increased
in
erythrocytes
GPx
activity
with
a
subsequent
decrease
in
plasma
TBARS
levels
but
anaerobic
training
had
no
effect
on
this
process.
Keywords:
superoxide
dismutase,
glutathione
peroxidase,
thiobarbituric
acid,
reactive
substances,
exercise,
aerobic
and
anaerobic
training,
non
sporting
men,
long-distance
runners,
antioxidant
enzyme
activities
Increased
energy
demand
during
physical
exercise,
especially
of
the
aerobic
type,
necessitates
a
multifold
increase
in
oxygen
supply
to
active
tissues.
During
exercise,
Correspondence
should
be
addressed
to
Berkant
Muammer
Kayatekin,
MD
Department
of
Physiology,
Dokuz
Eyliil
University
Medical
School
35340
Balgova,
Izmir,
Turkey
Fax:
00-90-232-2590541
E-mail:
kayabm@deu.edu.tr
0231-424X/2000/$
5.00
©
2000
Akademiai
Kiad6,
Budapest
268
Semih
Selamoglu
et
al.
bodily
oxygen
consumption
is
greatly
increased,
up
to
10-
to
15-fold
greater
than
resting
levels
[9].
Oxygen-centred
radicals
are
produced
in
the
intermediate
metabolism
[4].
Most
of
the
oxygen
consumed
in
the
mitochondria
is
utilized
to
produce
adenosine
5'-
triphosphate,
but
during
oxidative
phosphorylation
the
superoxide
radicals,
hydrogen
peroxide,
and
hydroxyl
radicals
are
produced
by
the
univalent
reduction
of
oxygen
and
leak
out
of
the
electron
transfer
chain
[6].
As
a
whole,
they
are
classified
as
reactive
oxygen
species
(ROS)
and
are
responsible
for
a
series
of
biochemical
and
physiological
changes,
namely
oxidative
stress.
The
ROS
released
cause
the
lipid
peroxidation
of
polyunsaturated
fatty
acids
in
the
biological
membranes
and
blood,
inducing
alterations
of
the
cell
functions
[7].
Lipid
peroxides
readily
decompose
to
liberate
highly
reactive
carbonyle
fragments
such
as
malondialdehyde.
Malondialdehyde
(MDA)
was
the
major
species
responsible
for
thiobarbituric
acid
reactive
substances
(TBARS)
[8].
Strenuous
physical
exercise
induces
oxidative
damage
to
lipids
in
various
tissues
[22,
23].
In
resting
state
the
body
is
equipped
with
both
non-enzymatic
and
enzymatic
antioxidant
reserves
to
prevent
the
potentially
harmful
effects
of
ROS
[13].
The
fine
physiological
balance
between
oxidative
reactions
and
antioxidant
capacity
may
be
perturbed
by
intense
physical
activity.
Antioxidant
defence
systems
preserve
homeostasis
for
normal
cell
function
at
rest
and
perhaps
during
mild-oxidative
stress.
Primary
components
of
the
physiological
antioxidant
defence
are
superoxide
dismutase
(SOD),
catalase
and
glutathione
peroxidase
(GPx).
SOD
catalyzes
the
dismutation
of
superoxide
to
0
2
and
H
2
0
2
,
which
catalase
(CAT)
converts
to
water
and
0
2
.
GPx
can
reduce
H
2
0
2
to
form
glutathione
disulphide
and
water
[5].
Large
number
of
studies
have
tested
the
effect
of
a
variety
of
endurance
exercise
training
regimens
on
antioxidant
defences,
but
information
on
the
effect
of
anaerobic
training
on
antioxidant
defences
is
scanty.
The
aim
of
the
present
study
was
to
assess
the
effects
of
aerobic
and
anaerobic
training
on
serum
lipid
peroxidation
levels
and
on
antioxidant
enzymatic
activities
in
erythrocytes.
Materials
and
Methods
This
study
consisted
of
33
non-smoking
males
(17.6102.28
years,
BMI
21.94
0.63);
including
11
long-distance
runners
(aerobic
training
group)
and
11
wrestlers
(anaerobic
training
group)
who
have
been
doing
sport
for
average
5.18
0.70
years
on
the
awerage
and
in
control
group.
The
control
subjects
did
not
perform
any
regular
physical
activity
before
the
study.
All
the
experiments
were
performed
in
according
to
the
Helsinki
Declaration.
Subjects
attended
the
laboratory
in
the
morning,
after
a
12
h
fast
and
a
10
ml
blood
sample
was
obtained
from
an
antecubital
vein.
Blood
samples
were
collected
48
Acta
Physiologica
Hungarica
87,
2000
Aerobic
and
anaerobic
training
and
antioxidant
enzymes
269
hours
after
the
termination
of
the
training
in
order
to
minimise
the
residual
effect
of
the
last
exercise.
Heparinized
venous
whole
blood
was
used
for
measuring
erythrocyte
antioxidant
enzymes
(SOD
and
GPx)
activities.
Blood
hemoglobin
values
for
calculating
enzyme
activities
were
determined
by
using
Coulter
Counter.
Plasma
TBARS
levels
were
measured
as
an
indicator
of
lipid
peroxidation.
Determination
of
SOD
activity
Erythrocyte
SOD
was
determined
with
a
Randox
test
combination
(Randox,
Crumlin,
UK).
Xanthine
and
xanthine
oxidase
were
used
to
generate
superoxide
radicals
reacting
with
2-(4-iodophenyl)
3-(4-nitrophenol)-5
phenyl
tetrazolium
chloride
(INT)
to
form
a
red
formazan
dye.
The
concentration
of
the
substrates
were
0.075
mol
for
xanthine
and
0.037
mmol
for
INT.
Superoxide
dismutase
inhibits
this
reaction
by
converting
the
superoxide
radical
to
oxygen.
A
SOD
unit
inhibits
the
rate
of
reduction
of
INT
by
50%
in
a
complex
system
with xanthine
and
xanthine
oxidase.
Because
of
the
small
linearity
range
of
the
test,
the
sample
was
diluted
so
that
the
percentage
of
inhibition
fell
between
30%
and
60%.
A
standard
curve
was
prepared,
using
the
kit
standard,
and
the
value
for
the
diluted
sample
was
read
from
this
curve.
SOD
activity
was
measured
at
505
nm
on
a
Shimadzu
UV-1201v
spectrometer
on
hemolysates
of
washed
erythrocytes
obtained
by
centrifugation
of
whole
blood.
Results
were
expressed
in
SOD
U/g
hemoglobin.
Determination
of
GPx
activity
GPx
was
also
determined
with
a
Randox
test
combination
(Randox,
Crumlin,
UK).
GPx
catalyses
the
oxidation
of
glutathione
(at
a
concentration
of
5
mmol)
by
cumene
hydroperoxide
according
to
the
method
of
Paglia
and
Valentine
[21].
In
the
presence
of
glutathione
reductase
(at
a
concentration
>0.75
10-3
U)
and
0.35
mmol
NADPH,
the
oxidised
glutathione
was
immediately
converted
to
the
reduced
form
with
a
concomitant
oxidation
of
NADPH
to
NADP
±
.
The
decrease
in
absorbance
at
340
nm
was
measured
at
37
°C.
The
assay
was
performed
on
a
hemolysate
of
washed
erythrocytes
obtained
from
the
mixing
of
0.05
ml
whole
blood
with
1
ml
cold
diluting
agent
and
1
ml
Drabkin
reagent.
The
GPx
unit
was
defined
as
the
enzyme
activity
necessary
to
convert
1
mmol
of
NADPH
to
NADP
in
1
minute.
The
activity
of
GPx
is
expressed
in
U/g
hemoglobin.
Acta
Physiologica
Hungarica
87,
2000
270
Semih
Selamoglu
et
al.
Determination
of
TBARS
level
In
a
modified
Yagi
method
[24],
0.05
ml
of
blood
was
sampled
with
a
pipette
for
determination
of
blood
cells
and
placed
in
1.0
ml
of
normal
saline
in
a
centrifuge
tube.
After
gently
shaking,
the
tube
was
spun
at
3000
rpm
for
10
minutes
and
0.5
ml
of
the
supernatant
was
transferred
to
another
centrifuge
tube.
The
4.0
ml
of
1/12
N
H
2
SO
4
was
added
to
this
solution
and
the
mixture
was
shaken
gently.
Then
0.5
ml
of
10%
phosphotungstic
acid
was
added
and
mixed.
After
standing
at
room
temperature
for
5
minutes,
the
mixture
was
centrifuged
at
3000
rpm
for
10
min.
After
the
supernatant
was
discarded,
the
sediment
was
mixed
with
2.0
ml
of
N/12
H
2
SO
4
and
0.3
ml
of
10%
phosphotungstic
acid
and
the
mixture
was
centrifuged.
The
sediment
was
suspended
in
4.0
ml
of
distilled
water
and
1.0
ml
of
thiobarbituric
acid
(TBA)
reagent
was
added.
The
reaction
mixture
was
heated
for
60
min
at
100
°C
in
a
water
bath.
After
cooling
with
tap
water,
5.0
ml
of
n-butanol
was
added
and
the
mixture
was
shaken vigorously,
then
centrifuged
at
3000
rpm
for
15
min.
Finally,
the
n-butanol
layer
was
taken
for
spectophotometric
measurement
at
532
nm.
A
standard
curve
was
prepared
using
the
MDA
standard
(1,
1,
3,
3-tetraethoxypropane)
and
the
value
for
the
plasma
was
read
from
this
curve.
The
results
were
expressed
as
nmol/ml.
Determination
of
VO
2
max
VO
2
max
was
determined
indirectly
by
Astrand's
method
using
Monarck
bicycle
ergometer
[3].
Statistical
analysis
All
results
were
expressed
as
mean
S.E.M.
The
statistical
analysis
of
the
data
was
performed
using
Mann—Whitney
U
test.
The
significance
was
set
at
p<0.05.
Results
When
the
effect
of
exercise
on
aerobic
power
was
examined;
indirect
VO
2
max
of
long-distance
runners
were
found
higher
than
wrestlers
and
non-sporting
men
(respectively,
p<0.001,
p<0.001,
Table
I).
Acta
Physiologica
Hungarica
87,
2000
Aerobic
and
anaerobic
training
and
antioxidant
enzymes
271
Table
I
Indirect
VO
2
max
values,
SOD,
GPx
activities
and
TBARS
levels
of
sportsmen
and
of
the
control
group
Groups
VO
2
max
SOD
GPx
TBARS
(ml/kg/min)
(U/gHb) (U/gHb)
(nmol/ml)
Control
47.40
01.57
1150.36
0
51.60
42.96
0
4.86
1.43
0
0.09
Distance
runners
62.30
01.85***
1059.27
0111.29
56.62
0
3.68**
1.08
0
0.10*
Wrestlers
49.71
01.49
1269.61
0
66.44
45.54
0
3.13
1.15
0
0.15
Results
were
presented
as
means
0S.E.M.
*
Lower
than
control
p<0.05
**
Higher
than
wrestlers
p<0.05
***
Higher
than
wrestlers
and
control
p<0.001
When
plasma
lipid
peroxidation
levels
and
antioxidant
enzymatic
activities
were
investigated;
TBARS
levels
of
long-distance
runners
were
lower
than
those
of
non-
sporting
men
(p<0.05)
but
similar
to
those
measured
in
wrestlers.
GPx
enzyme
activity
of
long-distance
runners
was
higher
than
those
of
wrestlers
(p<0.05)
but
similar
to
those
found
in
the
non-sporting
males.
There
were
no
significant
difference
among
the
groups
in
SOD
enzyme
activities
(Table
I).
A
positive
correlation
between
the
subject's
VO
2
max
and
GPx
activity
was
found
(r=0.53,
p<0.01).
A
negative
correlation
between
the
subject's
VO
2
max
and
plasma
MDA
level
was
found
(r=-0.38,
p<0.05).
Discussion
Increased
oxygen
utilization
during
exercise
cause
generation
of
free
radicals
[11].
Chronic
aerobic
training
has
been
claimed
to
reduce
exercise-increased
lipoperoxidation
by
improving
the
body's
defence
capabilities
against
free
radicals
generation,
likely
as
a
result
of
an
adaptive
increase
in
the
activities
of
the
scavenger
enzyme
systems
[2].
Scientific
literature
on
the
effect
of
anaerobic
training
on
antioxidant
defence
system
and
on
lipid
peroxidation
is
scanty
[17].
Human
erythrocytes
are
well
equipped
with
the
enzymes
SOD,
catalase
and
GPx,
that
protect
the
cells
against
the
accumulation
of
superoxide
radical
and/or
hydrogen
peroxide
normally
produced
during
the
oxidation
of
hemoglobin
[19].
It
has
been
reported
that
endurance
training
elevates
the
antioxidant
enzyme
activities
in
blood
at
rest
and
during
post-exercise
recovery
[14,
17].
However,
a
controversy
still
exists
as
to
which
enzyme
and
under
what
condition
an
enzyme
can
be
activated
[11,
12, 15,
16].
Available
data
suggest
that
each
of
the
antioxidant
systems
may
have
a
different
Acta
Physiologica
Hungarica
87,
2000
272
Semih
Selamoglu
et
al.
response
to
acute
and
chronic
exercise
depending
upon
their
biochemical
and
molecular
mechanism
of
regulation
[10].
Mena
et
al.
described
that
under
resting
conditions
the
SOD
and
GPx
activities
were
higher
in
cyclists
than
in
the
control
group
[18].
Marzatico
et
al.
determined
that
blood
SOD
and
GPx
activities
were
higher
in
marathon
runners
and
sprinters
than
in
controls
[17].
Ohno
et
al.
have
reported
that
in
sedentary
students
after
a
brief
physical
exercise
no
increase
in
erythrocyte
SOD
activity
was
found
[19].
In
another
study,
resting
blood
SOD
and
GPx
activities
were
no
different
in
jump-trained
(volleyball
players)
compared
with
untrained
subjects
[20].
It
has
been
reported
that
blood
GPx
activity
increased
after
swim
training
program
in
mice
[14].
The
results
from
the
present
investigation
demonstrated
that
SOD
activity
was
unaffected
by
aerobic
and
anaerobic
training.
GPx
enzyme
activities
of
long-distance
runners
were
higher
than
those
detected
in
wrestlers.
Despite
exercise-induced
free
radical
changes,
there
is
a
positive
side
to
oxidative
stress
associated
with
regular
exercise
[1].
Subjects
with
high
aerobic
power
show
significantly
greater
antioxidant
enzyme
activity
[17,
23].
It
has
been
reported
that
there
exist
a
good
correlation
between
exercise
endurance
time
and
GPx
activity
[1].
In
this
study,
a
positive
correlation
between
the
subject's
VO
2
max
and
GPx
activity
was
found.
This
could
further
show
how
even
aerobic
training
is
able
to
prevent
the
toxic
effects
of
lipid
peroxidation.
MDA
is
the
end
product
of
lipid
peroxidation
and
is
a
well-known
parameter
for
determining
the
increased
free
radical
formation
in
the
body.
It
has
been
reported
that
lipid
peroxidation
levels
are
lower
in
endurance-trained
than
in
untrained
animals
[2].
The
knowledge
on
the
effect
of
training
on
lipid
peroxidation
in
humans
is
sparse
equivocal.
Ohno
et
al.
[19]
and
Jenkins
et
al.
[12]
determined
that
plasma
MDA
levels
were
decreased
related
to
training
adaptation.
However,
Marzatico
et
al.
found
that
resting
plasma
MDA
levels
of
marathon
runners
and
sprinters
are
higher
than
control
levels
[17].
Results
of
studies
on
lipid
peroxidation
induced
by
exercise
are
actually
inconsistent
due
to
the
wide
variety
of
methods
employed
and
the
differences
in
exercise
protocols
(e.g.
type,
duration
and
intensity
of
exercise).
It
was
observed
that
resting
plasma
MDA
negative
by
correlated
with
the
aerobic
capacity
of
the
individuals,
suggesting
a
protective
effect
of
physical
fitness
[23].
In
the
present
study,
when
resting
plasma
lipid
peroxidation
levels
were
examined;
a
negative
correlation
between
the
subject's
VO
2
max
and
plasma
TBARS
level
was
found.
TBARS
levels
of
long
distance
runners
were
lower
than
control
levels
but
TBARS
levels
of
wrestlers
did
not
differ
from
those
found
in
controls.
In
conclusion,
these
results
suggest
that
aerobic
training
increased
GPx
activity
in
erythrocytes
with
a
subsequent
decrease
in
plasma
TBARS
levels
while
anaerobic
training
had
no
effect
on
this
process.
Acta
Physiologica
Hungarica
87,
2000
Aerobic
and
anaerobic
training
and
antioxidant
enzymes
273
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