Bali Medical Journal (Bali Med. J.) 2015, Volume 4, Number 1: 32-36
P-ISSN.2089-1180, E-ISSN.2302-2914

XANTHINE OXYDASE INHIBITION OF KOMBUCHA TEA
IN HYPERURICEMIA INDUCED WISTAR RAT: decrease of uric acid,
malondialdehyde, and 8-hydroxy-2'-deoxyguanosine
I Dewa Made Sukrama
Faculty of Medicine Udayana University, Bali-Indonesia

Background: Hyperuricemia is a condition of high level of uric acid in the body due to distortion of
purine nucleoside metabolism through hipoxanthin, xanthin, and guanin of basic purine. Objective: to
find a cure of hyperuricemia base on the utilization of kombucha tea. Methods: This is a true
experimental study by applying posttest only control group design to determine whether kombucha tea
inhibit xanthine oxidase in hyperuricemic induced rat reveales by decrease of uric acid,
malondialdehyde (MDA), and 8-hydroxy-2’-deoxyguanosine (8-OHdG). In this study, hyperuricemia
rat was achieved by intake of high purine diet. Rats were fed with a mixture of 4 g/kg BW of Gnetum
gnemon with 50 mL/kg BW of chicken liver ad libitum for 9 days. Treatments in this research are
combination of fermentation time of Kombucha tea and volume of this tea, i.e fermentation time 4, 8,
and 12 days and the volume are 1 mL and 4 mL. Therefore, there would be seven groups of treatment
including control group. ANOVA was then applied to determine the treatment effect with p < 0.05 was
concidered significant. Results: This study indicates that kombucha tea has an ability to inhibit
xanthine oxidase in hyperuricemic induced rat and decrease uric acid, MDA, and 8-OHdG. This
ability was achieved with combination treatment of 12 days fermentation and 4 mL of kombucha
intake. Xanthine oxidase, uric acid, MDA, and 8-OHdG levels by this treatment were obtained
significantly lower compare to control group. Conclusion: This study proved that kombucha tea was
potent to cure hyperuricemia of wistar rat via inhibition of xanthine oxidase produced.
Keywords: hyperuricemia; rat; kombucha tea; xanthine; oxidase.

INTRODUCTION
Uric acid is a metabolic product of exogenous
(brought in with food) or endogenous purine bases.
This acid in most physiologic fluids is an end
product of purine degradation. The serum urate
level in a given patient is determined by the amount
of purines synthesized and ingested, the amount of
urate produced from purines, and the amount of
uric acid excreted by the kidney (and, to a lesser
degree, from the gastrointestinal tract).1,2 Gout is an
inflammatory arthritis caused by the deposition of
monosodium urate crystals in tissues.1 This
condition typically occurs after years of sustained
hyperuricemia. It is estimated to affect 5.1 million
people in the United States according to the most
recent National Health and Nutrition Examination
Survey (NHANES III).2 Gout affects approximately
2% of men older than 30 years and 2% of women
older than 50 years, and is the most common form
of inflammatory joint disease in men older than 40
years. Serum uric levels are, on average, 0.5 to 1.0
mg/dl higher in men than women, making male sex

a risk factor for hyperuricemia and gout. Lower
serum uric levels in women are associated with the
presence of estrogen, which is thought to act as an
antihyperuricemic.3 In Indonesia, based on Health
Survey in the year of 2005, there were around 1020% men and post menopause women have a
higher levels of uric acids than normal person.4 It
was proven that, celery seed is often used in
treating this condition, as it possesses many antiinflammatory compounds. Other helpful herbs
include turmeric, boswellia, cayenne, colchicum
and hyssop were also potent to treat hyperuricemia.
Clearly, uric acid is produced by purine nucleoside
metabolism through hipoxanthin, xanthin, and
guanin basic purine. Distortion of this metabolism
leads to elevate level of uric acid and known as
hyperuricemia.5 Kombucha tea is a traditional
fermented tea, empirically in Balinese traditional
medicine was proven as a cure of hyperuricemia.
This study was carried in order to investigate the
ability of kombucha tea to inhibit further uric acid
formation in the hyperuricemia wistar rat.

Address of Correspondence: I D. M. Sukrama
Faculty of Medicine Udayana University
Bali-Indonesia
Email: dewa_sukrama@yahoo.co.id

METHODS
This is an experimental study with posttest
only control group design to observe inhibition
activity of kombucha tea in hyperuricemia induced
rat. A number of 28 wistar rats were employed in

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Bali Medical Journal (Bali Med. J.) 2015, Volume 4, Number 1: 32-36
P-ISSN.2089-1180, E-ISSN.2302-2914

this study and divided into 7 groups, i.e. control
group (C), the first group (T1) treated with 1 ml of
4 days fermented kombucha tea, the second group
(T2) treated with 1 ml of 8 days fermented
kombucha tea, the third group (T3) treated with 1
ml of 12 days fermented kombucha tea, the fourth
group (T4) treated with 4 ml of 4 days fermented
kombucha tea, the fifth group (T5) treated with 4
ml of 8 days fermented kombucha tea, and the sixth
group (T6) treated with 4 ml of 12 days fermented
kombucha tea. Before treatment rats were induced
to hyperuricemia by feeding with a mixture of 4
g/kg BW of Gnetum gnemon with 50 mL/kg BW of
chicken liver ad libitum for 9 days.
Animal ethical clearance was obtained from a
local authority body at Veterinary Faculty Udayana
University, Bali-Indonesia. Around 1 mL of blood
was taken from rat heart aorta which was anesthesia
before proceeding. The blood was then centrifuged
for 15 minutes at the rate of 3.000-3.500 rpm. Uric
acid reagent, FS TBHBA (2,4,6-tribromo-3hydroxybenzoic acid) was then added to the serum
obtained. The mixture was then incubated for 10
minutes at a temperature of 370C. Then, optical
density of the mixture was determined using
sphectrophotometer at 546 nm of wave number.
Anova was performed to determine the
different effect amongst treatment with p<0.05 was
considered significant.
RESULTS
Decrease of uric acid of hyperurisemic rat
Based line data of uric acid of hyperurisemia
rat were presented in Table 1.
Data of uric acid decrease of hyperurisemic rat
were presented in Table 2. Anova one way was then
applied to determine the treatment different. The
data were presented in Table 3
Table 1
Uric Acid Levels of Hyperurisemia Wistar Rat
No

Group
1
2
3
4
5
6
7

C
T1
T2
T3
T4
T5
T6
p**

Day-6th
2.63±0.10
2.76±0.07
2.72±0.04
2.60±0.06
2.70±0.05
2.94±0.54
3.17±0.58
0.083

Uric acid (mg/dL)
p*
Day-9th
0.688
3.79±0.19
0.584
4.06±0.86
0.714
4.11±0.83
0.473
3.59±0.59
0.329
3.93±0.91
0.067
4.10±0.11
0.070
4.11±0.09
0.064

p*
0.063
0.842
0.751
0.092
0.322
0.102
0.068

C = control, T = treatment, p* significance for
normality data (normally distributed at p>0.05). p**
significance for homogenous of variance
(homogenous at p>0.05)
Decrease of xanthine oxidase on hyperurisemic
rat
Xanthine oxidase ativity data were determined
as IC50 (inhibitory concentration) and presented in
Table 4. One way Anova was employed to

determine the difference of xanthine oxydase
activity among treatment group and control group.
Resume of the analysis was presented in Table 5.
Table 2
Average of Uric Acid Concentrattion on
Hyperurisemic Rat On day-14th and day-19th after
treatment
No.

Groups

1
2
3
4
5
6
7

C
T1
T2
T3
T4
T5
T6
p**

Day-14th
4.85±0.69
3.86±0.94
4.11±0.83
3.52±0.56
3.87±0.89
3.91±0.31
4.06±0.09
0.137

Uric acid (mg/dL)
p*
Day-19th
0.411
5.74±0.56
0.526
3.64±0.70
0.756
3.98±0.88
0.075
3.37±0.52
0.310
3.71±1.01
0.076
3.48±0.27
0.149
2.08±0.18
0,068

p*
0.536
0.487
0.855
0.445
0.527
0.125
0.396

p* significance for normality data (normal at p >
0.05).p** significance for homogenous of variance
(homogenous at p > 0.05)
Tabel 3
Resume of Anova One Way of Uric Acid of
hyperurisemic rat on day-14th and day-19th after
treatment
Groups
C

T1

T2

T3

T4
T5

T1
T2
T3
T4
T5
T6
T2
T3
T4
T5
T6
T3
T4
T5
T6
T4
T5
T6
T5
T6
T6

Different of uric acid levels (mg/dL)
Day-14th
p*
Day-19th
p*
0.99
0.053
2.10
0.001
0.74
0.143
1.76
0.001
1.32
0.012
2.37
0.001
0.98
0.057
2.03
0.001
0.94
0.066
2.26
0.001
0.78
0.120
3.66
0.001
0.26
0.603
0.33
0.479
0.33
0.499
0.28
0.557
0.02
0.976
0.07
0.881
0.05
0.915
0.16
0.728
0.21
0.672
1.57
0.003
0.59
0.237
0.61
0.202
0.24
0.625
0.26
0.575
0.20
0.679
0.49
0.295
0.48
0.923
1.90
0.001
0.35
0.480
0.35
0.462
0.39
0.434
0.11
0.810
0.54
0.276
1.29
0.011
0.04
0.939
0.23
0.619
0.19
0.694
1.64
0.002
0.16
0.752
1.41
0.006

C = control, T = treatment, p* significance at p <
0.05.
Decrease of MDA Plasma of Hyperurisemic Rat
Data of MDA levels of hyperurisemis rat were
presented in Table 6. Then, one way Anova was
employed to determine the different of MDA
decrease among control and treatment groups.
Resume of the one way Anova was presented in
Table 7.
Decrease of 8-OHdG serum of Hyperurisemic
Rat
Data of 8-OHdG levels of hyperurisemic rat
were presented in Table 8. One way Anova was

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Bali Medical Journal (Bali Med. J.) 2015, Volume 4, Number 1: 32-36
P-ISSN.2089-1180, E-ISSN.2302-2914

then applied to determine the difference between 8OHdG among control and treatment groups.
Resume of the results were presented in Table 9.
Table 4
IC50 of Xanthin Oxydase in Hyperuricemic Rat
At day-14th and Day-19th after treatment
No
Group
.
1
C
2
T1
3 3 T2
4
T3
5
T4
6
T5
7
T6
p**

Xanthine Oxydase (ng/dL)
Day-14th
p*
Day-19th
55.98±1.95
0.334
56.24±1.25
54.12±0.69
0.489
53.62±0.65
54.08±0.85
0.761
53.97±0.82
53.52±0.56
0.475
53.37±0.52
53.87±0.89
0.310
53.71±1.01
53.91±0.31
0.403
53.48±0.27
54.09±0.58
0.059
42.63±0.55
0.157
0.278

Table 7
Resume of One Way Anova of MDA at Day-14th
and Day-19th after treatment
Groups
C

p*
0.572
0.438
0.859
0.445
0.527
0.125
0.068

T1

T2

C = control, T = treatment, p* significance for
normality data (normal at p > 0.05).
p**
significance for homogenous of variance
(homogenous at p > 0.05)
Table 5
Resume of One Way Anova of IC50 Xanthine
oxydase on day-14th and day-19th
Groups
C

T1

T2

T3

T4
T5

T1
T2
T3
T4
T5
T6
T2
T3
T4
T5
T6
T3
T4
T5
T6
T4
T5
T6
T5
T6
T6

Different of IC50 XOD (mg/dL)
Day-14th
p*
Day-19th
p*
1.86*
0.013
2.62*
0.001
1.91*
0.011
2.27*
0.001
2.46*
0.002
2.87*
0.001
2.11*
0.006
2.52*
0.001
2.07*
0.006
2.76*
0.001
1.88*
0.012
13.60*
0.001
0.04
0.951
0.36
0.534
0.59
0.394
0.25
0.661
0.25
0.721
0.10
0.867
0.21
0.762
0.14
0.809
0.02
0.977
10.98*
0.001
0.55
0.428
0.61
0.294
0.21
0.767
0.26
0.648
0.17
0.684
0.49
0.391
0.02
0.684
11.34*
0.001
0.35
0.617
0.35
0.546
0.39
0.580
0.11
0.843
0.58
0.410
10.73*
0.000
0.04
0.957
0.23
0.683
0.23
0.743
11.08*
0.001
0.19
0.784
10.84*
0.001

C = control, T = treatment, p* significance at p <
0.05.
Tabel 6
MDA of Hyperurisemic Rat at Day-14th and Day19th after treatment
No.

Groups

1
2

C
T1
T2
T3
T4
T5
T6
p**

3
4
5
6
7

Day-14th
1.68±0.02
1.66±0.04
1.65±0.04
1.63±0.06
1.63±0.04
1.59±0.06
1.36±0.08
0.488

MDA plasma (M/L)
p*
Day-19th
0.755
1.68±0.02
0.764
1.66±0.02
0.306
1.62±0.02
0.159
1.60±0.02
0.492
1.58±0.02
0.671
1.56±0.04
0.145
1.08±0.11
0.118

p*
0.783
0.577
0.177
0.880
0.572
0.192
0.152

C = control, T= treatment, p* significance of
normality data (normal at p > 0.05). p**
significance for homogeneous of variance
(homogenous at p > 0.05)

T3

T4
T5

T1
T2
T3
T4
T5
T6
T2
T3
T4
T5
T6
T3
T4
T5
T6
T4
T5
T6
T5
T6
T6

Difference of MDA (M/L)
Day-14th
p*
Day-19th
0.02
0.670
0.02
0.03
0.415
0.06
0.05
0.220
0.07*
0.04
0.257
0.09*
0.09*
0.024
0.12*
0.34*
0.001
0.58*
0.01
0.694
0.05
0.03
0.415
0.04
0.03
0.472
0.08*
0.07
0.059
0.10*
0.33*
0.001
0.57*
0.02
0.670
0.01
0.01
0.743
0.02
0.06
0.125
0.05
0.31*
0.001
0.52*
0.01
0.921
0.04
0.04
0.257
0.06
0.29*
0.001
0.53*
0.05
0.220
0.03
0.29*
0.001
0.49*
0.25*
0.001
0.47*

p*
0.573
0.107
0.047
0.011
0.002
0.001
0.139
0.280
0.038
0.007
0.000
0.672
0.504
0.167
0.001
0.280
0.077
0.001
0.461
0.001
0.001

C = control, T= treatment, p* significance at p <
0.05.
Table 8
Data of 8-OHdG of Hyperurisemia Rat
At Day-14th and Day-19th after treatment
No.

Groups

1
2
3
4
5
6
7

C
T1
T2
T3
T4
T5
T6
p**

Day-14th
0.62±0.03
0.64±0.01
0.65±0.01
0.63±0.03
0.61±0.02
0.58±0.01
0.56±0.01
0.135

8-OHdG (ng/mL)
p*
Day-19th
0.240
0.61±0.03
0.683
0.63±0.01
0.124
0.61±0.01
0.734
0.61±0.01
0.239
0.59±0.01
0.406
0.51±0.01
0.161
0.39±0.03
0.106

p*
0.238
0.272
0.406
0.161
0.124
0.972
0.962

C = control, T = treatment, p* significance for
normality data (normal at p > 0.05).
p**
significance for homogenous of variance
(homogenous at p > 0.05)
DISCUSIONS
This research indicated that there was decrease
of uric acid levels of hyperurisemic rat after intake
of kombucha tea. Treatment with 4 mL of 12 days
fermented kombucha gave the significance highest
decrease of uric acid. The decrease was 3.66 mg/dL
(p < 0.05) compare to control.
Setiawan and Suyono (2012) in their study
obtained that there was also decrease of uric acid of
hyperurisemic rat treated with kombucha. They
obtained that the highest decrease was 54% with a
dose of 8mL/d of 14 days fermented kombucha.
The advantages of kombucha have already
been reported widely. The target is not a certain
organ, instead of affecting all system of metabolism
in the body and detoxification. This will leads to
increase of endogenous immune system towards
any penetration of xenobiotic.
However,

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Bali Medical Journal (Bali Med. J.) 2015, Volume 4, Number 1: 32-36
P-ISSN.2089-1180, E-ISSN.2302-2914

mechanism of how its work has not been
understood completely.
Table 9
One Way Anove of 8-OHdG of Hyperurisemic Rat
at Day-14th and Day-19th after treatment
Groups
C

T1

T2

T3

T4
T5

T1
T2
T3
T4
T5
T6
T2
T3
T4
T5
T6
T3
T4
T5
T6
T4
T5
T6
T5
T6
T6

Difference of 8-OHdG (ng/mL)
Day-14th
p*
Day-19th
p*
0.03
0.080
0.02
0.114
0.03*
0.026
0.01
0.856
0.02
0.282
0.01
1.000
0.01
0.470
0.02
0.114
0.04*
0.012
0.08*
0.001
0.06*
0.001
0.22*
0.001
0.01
0.587
0.02
0.158
0.01
0.470
0.02
0.114
0.04*
0.018
0.05*
0.003
0.06*
0.001
0.09*
0.001
0.08*
0.001
0.24*
0.001
0.02
0.212
0.01
0.856
0.04*
0.005
0.03
0.081
0.07*
0.001
0.08*
0.001
0.09*
0.001
0.22*
0.001
0.03
0.080
0.02
0.114
0.05*
0.001
0.08*
0.001
0.07*
0.001
0.22*
0.001
0.03
0.056
0.05*
0.001
0.05*
0.003
0.19*
0.001
0.02
0.212
0.14*
0.001

C = control, T = treatment, p* significane at p <
0.05.
In this study, it was obtained that kombucha
tea decrease uric acid levels of hyperurisemic rat.
The uric acid levels decrease was followed by
decrease of xanthine oxydase. As indicates in Table
4, treatment of 4 ml of 12 days fermented
kombucha results the highest decrease, 2.76 mg/dl
compare to control and others treatments.
Therefore, it can be stated that the role of
kombucha tea in decreasing uric acid in
hyperurisemic rat is as an inhibitor the formation of
xanthine oxydase.
It was known that uric acid is produced during
catabolic of nucleotide purine through a process
catalyzed by xanthine oxyreductase (XOR) in the
liver process leads to hypoxanthine oxidation to
form xanthine and further oxidation forming uric
acid.12 During formation of uric acid, reactive
oxygen species was also produced which was
significantly increase vascular oxidative stress.12
XOR is an hepar enzyme that catalyze uric
acid, nitrous oxide, and reactive oxygen species
which were potent to damage deoxyribonucleic
acid, ribonucleic acid and its protein, inactivated
enzymes, amino acids oxidation, and peroxydation
of lipids.13 XOR can be present in two interconvertible form, i.e. XO-xanthine oxydase and
XDH-xhantine dehidrogenase.14
Increase of lipids peroxydation leads to
increase of free radical results in increase of
malondialdehyde formation as a marker of free
radical on blood. Increase of uric acid production

has also related to increase the formation of reactive
oxygen species and also increase of lipid
peroxidation. Lipid peroxidation produces MDA as
a marker of damaging membrane cell. In this study,
we observed the highest significant decrease of
MDA on treatment of 4 ml of 12 days fermented
kombucha tea. This also consistence to the data
obtained for uric acid and xanthine oxydase
decrease. In line with this finding, we also obtained
that there was a significant decrease of 8-OHdG, a
marker for DNA damage. The highest decrease was
also gained for treatment of 4 ml of 12 days
fermented kombucha tea. Increase of xanthine
oxydase on myocardial patients will also be
followed by increase of MDA. Since,
hyperuricemia can also lead to myocardial
destruction, therefore, this situation probably
happen in hyperuricemia patients.13
Deoxyguanosine (dG) is one of DNA
component and during oxidation form 8-hydroxy2'-deoxyguanosine (8-OHdG). Therefore, 8-OHdG
is a product of DNA oxidation cause by ROS.
Guanosine hydroxilation is a response of normal
metabolic process or can be due to other exogenous
factors. Not too many research regardless of the
role of kombucha tea in reducing increase of 8OHdG. The role of green tea in inhibition of DNA
damage was known through 8-OHdG increase on
mice expose with tobacco. They obtained that
obtained there was an inhibition of increase of 8OHdG.7
CONCLUSION
Role of kombucha tea in decreasing of uric
acid in hyperuricemia rat is through inhibition of
xanthine oxydase formation. This was proven by
decrease of uric acid which was followed by
decrease of xanthine oxydase.
After exposure phase, since kombucha tea is a
polar matter, no metabolism of phase-I occurs. The
tea will be absorbed straight forward and
distributed along with blood stream.
Pharmacodynamic
phase
occurs
with
formation of ligand in the hepar and followed by
decrease of xanthine oxydase formation.
Kombucha tea was also potent to decrease
MDA, a product of membrane cell damage due to
ROS which was formed during the production of
uric acid.
In this research, we also observed that there
was a decrease of 8-OHdG as a marker of DNA
damage.
AKCNOWLEDGMENT
The authors would like to thank Rector of
Udayana University and LPPM for the funding.
Thanks was also pointed to Prof I N Adiputra for
guidience during research. Thanks also to staff of
UPT Lab. Analitik Udayana University for access
and aid of their fasilities for managing the research.

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35

Bali Medical Journal (Bali Med. J.) 2015, Volume 4, Number 1: 32-36
P-ISSN.2089-1180, E-ISSN.2302-2914

Thanks also to Mr. Priono from Kristallindo for
providing reagents for uric analysis. And special
thank to Vetinery Board for providing rat for this
experiment.
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