J Med Sci, Volume 47, No.

2, June: 51-58
Lukman La Basy et al., The effects of the ethanolic extract of mahogany seeds
(Swietenia macrophylla King) on the renal function of streptozotocin-induced diabetic rats

The effects of the ethanolic extract

of mahogany seeds (Swietenia
macrophylla King) on the renal function of
streptozotocin-induced diabetic rats
Lukman La Basy1,*, Sri Lestari S R2, Sri Kadarsih2
1
Sekolah Tinggi Ilmu Kesehatan, Maluku Husada, Moluccas, Maluku,
2
Department of Physiology, Universitas Gadjah Mada, Yogyakarta, Indonesia

DOI: http://dx.doi.org/10.19106/JMedSci004702201501

ABSRACT
Diabetes-associated oxidative stress causes glomerular hypertrophy, decrease of glo-
merular filtration rate and inhibits cell proliferation that lead to the decrease of renal
function as indiated by the increase of serum creatinine level and the presence of
protein in urine. Mahogany seed (Swietenia macrophylla King) has been proven to have
antidiabetic activity. This study was conducted to evaluate the effect of the ethanolic
extract of mahogany seeds on the renal function of streptozotocin-induced diabetic rats.
Six normal rats as control (Group I) and 24 diabetic rats were used in this study. The
diabetic rats were randomized allocated into four groups with six rats in each group.
Group II was considered as diabetic rats control and received aquadest. Group III-V were
considered as extract administered diabetic group and received ethanolic extract of S.
macrophylla seed for 21 days at a dose of 50, 100 and 200 mg/kg BW, respectively.
Serum malondialdehyde (MDA), serum creatinine, and urine protein levels were monitored,
before and after the ethanolic extract of S. macrophylla seed administration. Serum MDA,
serum creatinine and urine protein levels of all rats after STZ induction (Group II-V) were
significantly higher than without STZ induction (p < 0.05). A significant decrease in the
serum MDA and serum creatinine as well as urine protein levels were observed after the
treatment with ethanolic extract of S. macrophylla seed compared to before treatment
(p < 0.05). In conclusion, the ethanolic extract of S. macrophylla seed is able to correct
renal dysfunction of streptozotocin-induced diabetic rats.

ABSTRAK
Stres oksidatif akibat diabetes dapat menyebabkan hipertropi glomerulus, penurunan
kecepatan filtrasi glomerulus dan penghambatan proliferasi sel yang berakibat terjadinya
penurunan fungsi ginjal sebagaimana ditunjukkan dengan penurunan kreatinin serum dan
adanya urin protein. Biji mahoni (Swietenia macrophylla King) terbukti mempunyai aktivitas
antidiabetes. Penelitian ini dilakuan untuk mengkaji efek ekstrak etanol biji mahoni pada
fungsi ginjal tikus diabetes yang diinduksi streptozotosin. Enam tikus normal sebagai
kontrol (Kelompok I) dan 24 tikus diabetes digunakan dalam penelitian. Tikus diabetes
dibagi menjadi 4 kelompok secara random dengan 6 ekor tikus masing-masing kelompok.
Kelompok II sebagai kontrol tikus diabetes menerima air suling. Kelompok III-V sebagai

Corresponding author: Lukman.apt@gmail.com

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J Med Sci, Volume 47, No. 2, June: 51-58

kelompok perlakuan menerima ekstrak etanol biji mahoni (S. macrophylla King) selama 21
hari dengan dosis berturut-turut 50, 100 dan 200 mg/kg BB. Kadar malodehaldehid (MDA)
serum, kreatinin serum dan protein urin diukur sebelum dan sesudah pemberian ekstrak
etanol biji mahoni. Kadar MDA serum, kreatinin serum dan protein urin pada semua tikus
setelah induksi STZ (Group II-V) lebih tinggi secara bermakna dibandingkan tanpa induksi
STZ (p<0,05). Penurunan secara nyata terhadap kadar MDA serum, kreatinin serum dan
protein urin dijumpai setelah pemberian ekstrak etanol S. macrophylla King dibandingkan
sebelum pemberian (p<0,05). Kesimpulan, ekstrak etanol biji mahoni dapat memperbaiki
disfungsi ginjal tikus diabetes yang diinduksi streptozotosin.

Key words : hyperglycemia, Swietenia macrophylla King, serum creatinine, MDA, urine
protein

INTRODUCTION rate and inhibits cell proliferation that lead to

Diabetes mellitus (DM) is a metabolic the decrease of renal function as indiated by
disease characterized by hyperglycemia due the increase of serum creatinine level and the
to a decrease in insulin secretion, insulin presence of protein in urine.5-7
action, or both.1 Complications of diabetes Previous studies showed the benefits and
due to hyperglycemia includes macrovascular potential of some medicinal plants in dealing
(coronary artery disease, peripheral arterial with oxidative stress in both the treatment
disease, and stroke) and microvascular of diabetes and its complication.8 Mahogany
(diabetic nephropathy, neuropathy, and (Swietenia macrophylla King) was proven
retinopathy) dysfunctions.2 Oxidative stress to have antidiabetic activity. The aqueous-
caused by hyperglycemia plays an important methanolic extract of the mahogany seed
role in those macrovascular and microvascular induces the pancreatic β cells regeneration
complications of DM.2,3 Oxidative stress and increases the insulin production due to
occurs as result of the increase of peroxidation its antioxidant activity through glucose auto-
product as a mediator of tissue damage and oxidant and protein glycation pathways.9 In
leads to the activation of pathogenesis addition, the alcoholic mahogany seed extract
complication pathways of DM.3 stimulates the glucokinase enzyme, a major
Malondialdehyde (MDA) is a product catalyst in the posporilation of glucose, in
of lipid peroxidation due to the increase of lowering blood glucose levels.10
reactive oxygen species (ROS) level on the Streptozotocin (STZ) is widely used to
oxidative stress conditions. Plasma MDA is induce experimental both insulin-dependent
frequently used as biomarker for oxidative and non-insulin-dependent diabetes mellitus
stress. In diabetics condition, plasma MDA in animals, respectively. The mechanism of
level increases due to the decrease of STZ action in β cells has been intensively
antioxidant level and leads the early systemic investigated and is well understood.11
complications of diabetes as expressed by Streptozotocin entering the B cell via a glucose
the increased AGEs, protein, lipid serum, transporter (GLUT2) causes DNA damage
lymphocytes, and urine of diabetic patients.4,5 due to alkylation of DNA. The DNA damage
Moreover, oxidative stress causes glomerular induces activation of poly ADP-ribosylation
hypertrophy, decrease of glomerular filtration leads to depletion of cellular NAD+ and ATP

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 Lukman La Basy et al., The effects of the ethanolic extract of mahogany seeds
(Swietenia macrophylla King) on the renal function of streptozotocin-induced diabetic rats

resulting in the formation of ROS. Moreover, freshly citrate buffer (0.01 M and pH = 4.5).
STZ liberates toxic amounts of nitric oxide The day of STZ injection was designated as
that inhibits aconitase activity and participates day 0 (D-0). Development of diabetes was
in DNA damage. As a result of the STZ action, confirmed by measuring blood glucose level
β cells undergo the destruction by necrosis.12,13 four days after STZ injection (D-4). Rats
The present study was designed to with the blood glucose level higher than 200
evaluate the effects of the ethanolic extract mg/dL were considered to be diabetic. The
of mahogany seeds (S. macrophylla King) on diabetic rats were then selected for the study.
the renal function of streptozotocin-induced This study has been approved by Medical and
diabetic rats. The renal function was expressed Health Research Ethics Committee, Faculty
by serum creatinine and urinary protein levels. of Medecine, Universitas Gadjah Mada,
Furthermore, the plasma MDA level after the Yogyakarta.
ethanolic administration was also evaluated.
Animal treatment
MATERIALS AND METHODS Six normal rats and 24 diabetic rats were
Ethanolic extract preparation used in this study. The normal rats were
considered as non diabetic rats control and
The S. macrophylla King seeds were
received aquadest (Group I). The diabetic
collected in The Bogor Botanical Gardens,
rats were randomized allocated into four
Indonesia and identified by a botanist in The
groups with six rats in each group. Group
Faculty of Pharmacy, Universitas Gadjah
II was considered as diabetic rats control
Mada Yogyakarta. The seeds were separated
and received aquadest. Group III-V were
from the fruits, dried under open air conditions
considered as extract administered diabetic
and powdered using a blender. The powdered
group and received ethanolic extract of S.
seeds were extracted with ethanol by the
macrophylla seed at a dose of 50, 100, and
ethanol method for three days. The extract
200 mg/kg BW, respectively. The ethanolic
was filtered through Whatman filter and the
extract was administered one time per day at
filtrate was collected and concentrated in a
08.00 AM started from day 4 and continued
rotary evaporator at 40 oC. The concentrated
for next 21 days. Blood glucose level was
extract was dried under open air and stored
measured and monitored on day 0 (D-0), 4
under refrigeration until further use.
(D-4), 18 (D-18) and 25 (D-25), respectively.
In addition, serum creatinine, serum MDA
Induction of diabetes by streptozotocin
and urine protein levels were also monitored,
Wistar strain male albino rats, 6-8 weeks of however only on day 4 (D-4) and 25 (D-25),
age, weighing from 180 to 200 g were selected respectively.
for this study. All rats were acclimated for a
period of 7 days in our laboratory condition Biochemical analysis
prior to the study. Rats were housed at room
A volume of 2 mL orbital sinus blood
temperature under 12 hours cycles of dark and
sample was collected from each study rat and
light and were fed standard food and water ad
assayed by standard methods for estimation
libitum. Induction of diabetes in the rats were
of blood glucose, serum creatinine and MDA
performed by a single intraperitoneal injection
levels. The blood glucose level was measured
of STZ at dose of 60 mg/kg BW in 0.1 mL of

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J Med Sci, Volume 47, No. 2, June: 51-58

by glucose oxidase peroxidase (GOP-PAP) Group II-V had the blood glucose level higher
method.14 The serum creatinine level was than 200 mg/dL (p > 0.05). Furthermore, a
measured by Jaffe method15, MDA level significant decrease in the level of blood
was measured by TBARS metod16 and urine glucose level was observed 14 (D-18) as
protein level was measured by colorimetric test well as 21 (D-25) days after the treatment
pyrogallon red metod.17 All spectrophotometric with ethanolic extract of S. macrophylla seed
measurements were carried out in a DR6000 (Group III-V) (p < 0.05). The blood glucose
UV-Visible spectrophotometer (Hach-US). levels in diabetic rats of the Group III-V were
in normal level (<200 mg/mL) on D-18 and
Statistical analysis D-25. In contrast, the blood glucose level in
All the results were expressed as mean diabetic rats of Group II (without treatment
± standard deviation (SD) for six animals with the ethanolic extract) remained higher on
in each group. Statistical comparisons were D-18 and D-25 (p > 0.05) (TABLE 1).
performed using one way analysis of variance
(ANOVA) followed by Post Hoc Tests Least The effect of ethanolic extract of maho-
Significant Difference (LSD). p-values of gany seeds on serum MDA level
less than 0.05 were considered to indicate Serum MDA level of all rats four days after
statistical significance. STZ induction (Group II-V) were significantly
higher than without STZ induction (p <
RESULTS 0.05) (TABLE 2). A significant decrease in
the serum MDA level was observed 21 (D-
The effect of ethanolic extract of mahogany
25) days after the treatment with ethanolic
seeds on blood
​​ glucose level
extract of S. macrophylla seed (Group III-V)
Blood glucose level of all rats in all groups compared to before treatment (D-4) (p <
before STZ induction were in normal level 0.05). No significant difference in the serum
(TABLE 1). A significant increase in the level MDA level was observed on D-4 compared to
of blood glucose level was observed four days on D-25 in Group II (p > 0.05) (TABLE 2).
after STZ induction (D-4) when compared to
before STZ induction (p < 0.05). All rats on

TABLE 1. Effect of ethanolic extract of S. macrophylla seed on blood glucose levels (mg/dL) in diabetic rats

Mean ± Standard Deviation

Group Treatment
D-0 D-4 D-18 D-25
I Aquadest 82.42 ± 5.24 77.75 ± 3.09 82.73 ± 5.04 84.90 ± 6.75
II STZ + aquadest 74.83 ± 1.81 230.10 ± 2.99 233.48 ± 6.06 234.15 ± 3.23
III STZ + extract 50 mg 74.12 ± 2.55 227.66 ± 8.69 183.28 ± 13.06 145.43 ± 5.83
IV STZ + extract 100 mg 75.76 ± 2.45 230.45 ± 3.74 146.07 ± 14.81 124.57 ± 2.06
V STZ + extract 200 mg 75.31 ± 3.28 230.86 ± 4.30 127.01 ± 23.55 102.92 ± 4.42

D-0: day on STZ induction; D-4: 4 days after STZ induction; D-18: 14 days after administration of ethaolic extract;
D-25: 21 days after administration of ethanolic extract.

54
 Lukman La Basy et al., The effects of the ethanolic extract of mahogany seeds
(Swietenia macrophylla King) on the renal function of streptozotocin-induced diabetic rats

TABLE 2. Effect of ethanolic extract of S. macrophylla seed on serum MDA level (mg/dL)
in diabetic rats

Mean ± Deviation Standard

Group Treatment
D-4 D-25
I Aquadest 2.27 ± 0.62 1.31 ± 0.13
II STZ + aquadest 8.72 ± 0.64 8.56 ± 0.44
III STZ + extract 50 mg 8.31 ± 0.77 5.23 ± 0.53
IV STZ + extract 100 mg 8.08 ± 0.47 3.71 ± 0.42
V STZ + extract 200 mg 8.22 ± 0.37 2.56 ± 0.35

D-4: 4 days after STZ induction; D-25: 21 days after administration of ethanolic extract.

The effect of ethanolic extract of mahogany observed 21 (D-25) days after the treatment
seeds on serum creatinine level with ethanolic extract of S. macrophylla seed
Serum creatinine level of all rats four (Group III-V) compared to before treatment
days after STZ induction (Group II-V) (D-4) (p < 0.05). No significant difference in
were significantly higher than without STZ the serum creatinine level was observed on
induction (p < 0.05) (TABLE 3). A significant D-4 compared to on D-25 in Group II (p >
decrease in the serum creatinine level was 0.05) (TABLE 3).

TABLE 3. Effect of ethanolic extract of S. macrophylla seed on serum creatinine level

(mg/dL) in diabetic rats

Mean ± Deviation Standard

Group Treatment
D-4 D-25
I Aquadest 1.26 ± 0.33 0.67 ± 0.04
II STZ + aquadest 3.49 ± 0.15 3.90 ± 0.23
III STZ + extract 50 mg 3.50 ± 0.26 2.12 ± 0.22
IV STZ + extract 100 mg 3.62 ± 0.18 1.37 ± 0.34
V STZ + extract 200 mg 3.56 ± 0.14 0.68 ± 0.09
D-4: 4 days after STZ induction; D-25: 21 days after administration of ethanolic extract.

Urine protein level of all rats four days after S. macrophylla seed (Group III-V) compared
STZ induction (Group II-V) were significantly to before treatment (D-4) (p < 0.05). No
higher than without STZ induction (p < 0.05) significant difference in the urine protein level
(TABLE 3). A significant decrease in the urine was observed on D-4 compared to on D-25 in
protein level was observed 21 (D-25) days Group II (p > 0.05) (TABLE 4).
after the treatment with ethanolic extract of

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J Med Sci, Volume 47, No. 2, June: 51-58

TABLE 4. Effect of ethanolic extract of S. macrophylla seed on urine protein level (mg/
dL) on diabetic rats

Mean ± Deviation Standard

Group Treatment
D-4 D-25
I Aquadest 2.27 ± 0.62 1.31 ± 0.13
II STZ + aquadest 8.72 ± 0.64 8.56 ± 0.44
III STZ + extract 50 mg 8.31 ± 0.77 5.23 ± 0.53
IV STZ + extract 100 mg 8.08 ± 0.47 3.71 ± 0.42
V STZ + extract 200 mg 8.22 ± 0.37 2.56 ± 0.35

D-4: 4 days after STZ induction; D-25: 21 days after administration of ethanolic extract.

DISCUSSION glucose levels.9 Moreover, Maiti et al.20 and

In this study, STZ induction resulted in Dewanjee et al.21 have successfully isolated
a significant increase in blood glucose level swietenine, a tetranortriterpenoid, from the
(Group II-V) compared to the control group S. macrophylla seeds and have proven its
(Group I). All rats on Group II-V had the hypoglycemic activity in type 2 diabetic rats.
blood glucose level higher than 200 mg/dL Serum MDA level of rats four days
indicating the rats became diabetes. Diabetic after STZ induction significantly higher
Streptozotocin-induced diabetes has been than without STZ induction indicating that
described a useful experimental model to oxidative stress developed in diabetic rats
study the activity of antidiabetic agents.18 after STZ induction. After administration of
Streptozoocin destroys β-cells of the pancreas ethanolic extract of S. macrophylla seed to
and induces hyperglycemia.11-13,19 the diabetic rats on Group III-V for 21 days,
After administration of ethanolic extract a significant decrease in serum MDA level
of S. macrophylla seed to the diabetic rats was observed (TABLE 2). It indicates that
on Group III-V for 21 days, a significant the ethanolic extract of S. macrophylla seed is
decrease in blood glucose level was observed able to correct the diabetes-induced oxidative
which was close to the normal level (< 200 stress. The ethanol extract of S. macrophylla
mg/dL). In addition, the antidiabetic effect of seeds was proven to have antioxidant activity
the ethanolic extract of S. macrophylla seed in the streptozotocin-induced diabetic rats.17
was in a dose-dependent manner (TABLE Furthermore, the antioxidaive compounds
1). The antidiabetic effect of S. macropylla from S. macropylla have been isolated such
seed extract has been previously reported by as swietemacrophyllanin, catechin, and
some authors.9,10,17 Its mechanism of action epicatechin.22
as antidiabetic has been also investigated. Serum creatinine and urine protein
Kalaivanan et al.10 reported that ethanolic level of rats four days after STZ induction
extract of mahogany seed can increase insulin significantly higher than without STZ
production and stimulate the regeneration of induction indicating the decrease of renal
β-cells of the pancreas. Swietenia macropylla function. Diabetes-associated oxidative stress
seed extract was also reported can activate causes glomerular hypertrophy, decrease of
the enzyme glucokinase in lowering blood glomerular filtration rate and inhibits cell

56
 Lukman La Basy et al., The effects of the ethanolic extract of mahogany seeds
(Swietenia macrophylla King) on the renal function of streptozotocin-induced diabetic rats

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