UNIVERSA MEDICINA

January-April, 2013 Vol.32 - No.1

Scurrula atropurpurea increases nitric oxide and de-

creases malondialdehyde in hypertensive rats

Nour Athiroh AS* and Erna Sulistyowati**

ABSTRACT

BACKGROUND *Department of Biology,

Hypertension is the most prevalent chronic disease and has an impact on one Faculty of Mathematics and
billion people. Production of superoxide radicals and endothelial dysfunction Natural Sciences, Islamic
are involved in hypertension. Scurrula atropurpurea (BL.) Dans. is a tea plant University of Malang, Malang,
East Java
parasite. This study aimed to evaluate the role of Scurrula atropurpurea (BL.)
**Department of Pathology,
Dans. on nitric oxide (NO) as a marker of endothelial dysfunction and
Faculty of Medicine, Islamic
malondialdehyde (MDA) as a marker of oxidative stress in hypertensive rats. University of Malang, Malang,
East Java
METHODS
This study subjected rats to deoxycorticosterone acetate (DOCA)-induced Correspondence
hypertension. The experimental groups consisting of the control group and 3 Nour Athiroh AS, S.Si., M.Kes.
hypertension groups receiving Scurulla atropurpurea extract at a dosage of Department of Biology,
50; 100; and 200 mg/KgBW. Scavenging activity of Scurrula atropurpurea Faculty of Mathematics and
(BL.) extract was analyzed by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. Natural Sciences, Islamic
The levels of arterial nitric oxide (NO) and pulmonary malondialdehyde (MDA) University of Malang,
Jl. Mayjen Haryono 193
were analyzed by spectrophotometry. ANOVA and a post hoc test were applied
Malang, East Java
to find the difference of arterial NO and pulmonary MDA levels between groups.
Phone: +62341-575461
Email:
RESULTS
[email protected]
The level of arterial NO was significantly decreased in the hypertension groups
as compared with the control group, while the level of pulmonary MDA was Univ Med 2013;32:44-50
significantly increased (p<0.05). Scurulla atropurpurea significantly increases
the NO level at a dosage of 200 mg/KgBW, compared with the hypertension
groups (p<0.001). Scurulla atropurpurea significantly decreases pulmonary
MDA level at a dosage of 100 and 200 mg/KgBW compared with the hypertension
groups (p<0.05).

CONCLUSION
Scurulla atropurpurea extract increases arterial NO and decreases pulmonary
MDA in hypertensive rats, thus playing an important role in endothelial
dysfunction and oxidative stress.

Key words: Mistletoe, arterial NO, pulmonary MDA, hypertension, rats

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Univ Med Vol. 32 No.1

Benalu teh meningkatkan kadar nitrit oksida dan menurunkan

kadar malondialdehid pada tikus hipertensi

ABSTRAK
LATAR BELAKANG
Hipertensi merupakan penyakit kronik yang paling kerap ditemukan dan memberikan pengaruh kepada sekitar satu
milyar individu. Hipertensi berhubungan dengan peningkatan produksi radikal superoksida dan disfungsi endotel.
Scurrula atropurpurea (BL.) Dans. merupakan tanaman parasitik yang menyerang teh. penelitian ini bertujuan untuk
mengevaluasi peranan benalu teh terhadap disfungi endotel yang duikur menggunakan kadar nitrit oksida (NO) dan
stres oksidatif diuukr menggunakan kadar mondialdehid (MDA) pada tikus model hipertensi.

METODE
Subyek penelitian adalah tikus model hipertensi yang diinduksi oleh deoxycorticosterone acetate (DOCA). Kelompok
perlakuan terdiri atas kelompok kontrol, kelompok tikus hipertensi, kelompok ekstrak benalu teh dosis 50; 100; dan
200 mg/kgBB. Analisis aktivitas scavenging dari benalu teh dilakukan dengen metode DPPH. Analisis kadar NO
arteri dan kadar MDA paru dilakukan dengan metode kolorimetrik dengan spektrofotometer. Uji ANOVA dan Post
Hoc dilakukan untuk melihat perbedaan kadar NO arteri dan MDA paru pada berbagai kelompok perlakuan.

HASIL
Terdapat penurunan kadar NO arteri dan peningkatan kadar MDA paru secara bermakna pada tikus hipertensi
dibandingkan kontrol (p<0,05). Ekstrak benalu teh dosis 200 mg/kgBB dapat meningkatkan kadar NO arteri secara
bermakna dibandingkan tikus hipertensi (p<0,001). Ekstrak benalu teh dosis 100 dan 200 mg/kgBB dapat menurunkan
kadar MDA paru secara bermakna dibandingkan tikus hipertensi (p<0,05).

KESIMPULAN
Ekstrak benalu teh meningkatkan kadar NO arteri dan menurunkan kadar MDA paru pada tikus hipertensi. Jadi
berperan penting pada disfungsi endotel dan stress oksidatif.

Kata kunci: Benalu teh, NO arteri, MDA paru, tekanan darah tinggi, tikus

INTRODUCTION
Various lines of evidence reveal the
involvement of reactive oxygen species and
Hypertension is the end result of a complex
oxidative stress in hypertension and the
interaction between genetic and environmental
development of its complications. Hypertension
factors affecting the physiological systems
is associated with increased production of
regulating blood pressure. Throughout the world
superoxide radicals and endothelial
hypertension is the most frequently encountered
dysfunction. (3) Superoxide radicals have a
chronic disease and affects around one billion
negative effect on endothelial function by
individuals.(1) Death resulting from hypertension
reacting directly with nitric oxide (NO), such that
may be caused by cerebrovascular and
they decrease NO bioavailabily. In addition,
cardiovascular complications, such as stroke,
peroxynitrites as the products of the reaction of
end-stage renal disease, congestive heart failure,
superoxide radicals with NO, also have negative
myocardial infarction, and cardiac standstill.(2)
effects on endothelial cells.(4,5) Hydroxyl radicals

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Athiroh, Sulistyowati Scurrula atropurpurea and hypertension

produced by the decomposition of were assigned randomly into the groups, with
hydroperoxynitrites may trigger lipid each group containing five rats.
peroxidation, marked by increased
malondialdehyde (MDA) levels. Preparation tea parasite crude extract
For the treatment of hypertension and its Botanical determination of the leaves of the
complications, many drugs of plant origin have tea parasite was done at the Indonesian
been developed, comprising digitoxin from Scientific Institute (LIPI) at Purwodadi,
Digitalis purpurea, reserpine from Rauwolfia Pasuruan, East Java. The leaves were washed,
serpentina, aspirin from Salix alba, left to dry in an oven at 40-600C, then ground
tetramethylpyrazine from Jathropha into a powder. One hundred milligrams of tea
podagrica, and tetrandrine from Stephenia parasite leaf powder was steeped in methanol
tetradra.(2) Scurrula atropurpurea (BL.) Dans. in an erlenmeyer flask of 1 L capacity. The
is a parasitic plant attacking tea plants and mixture was shaken for 30 mnutes to distribute
therefore known as the tea parasite. In the powder uniformly in the methanol.
Indonesia, especially on the island of Java, the Subsequently the mixture was left to stand
stems and leaves of this plant have been overnight until a precipitate was formed. The
traditionally used, among others for the treatment supernatant, being a mixture of methanol and
of cancers.(6) No studies have been found for the active constituents, was subjected to
evaluating the potential of the tea parasite for evaporation. The extract was labelled and stored
the treatment of hypertension. Therefore the in a freezer.(8) The TPLE was administered daily
objective of the present study was to evaluate by the oral route using a catheter, this being
the effect of tea parasite leaf extract (TPLE) continued for 6 weeks.
on the levels of NO as marker of endothelial
dysfunction and on the levels of MDA as a Determination of antioxidant activity
marker of oxidative stress in a rat model of The antioxidant activity of the extract was
hypertension. analyzed by a modification of the DPPH free
scavenging activity method.(9,10) The sample was
METHODS dissolved in methanol (at concentrations of 10-
100 ppm), reacted with 0.2 mM DPPH, and
Study design incubated for 30 minutes at room temperature.
This was an laboratory experimental study Absorbance was measured at 515 nm.
conducted from February to July 2012. Antioxidant activity was calculated as a
percentage of inhibition of DPPH (percentage
Preparation of rat hypertension models of scavenging effect), i.e. % inhibition = [1-
The study subjects were Wistar rats aged (absorbance of sample /absorbance of blank)]
3-5 months and weighing 250-300 grams. The x 100%. The IC50 is the concentration of the
rats were injected subcutaneously with sample required for yielding 50% inhibition.
deoxycorticosterone acetate (DOCA) at a
dosage of 10 mg/KgBW, 2 times weekly for 6 Determination of arterial NO and pulmonary
weeks. The rats were given 2% NaCl instead MDA
of drinking water. The blood pressure and the The specimens used for the analysis of
weights of the rats were then determined.(7) The NO levels were arterial tissues from the tails of
treatment groups consisted of the control group, the rats. The method used was the Griess
the group of non-TPLE hypertensive rats, three reaction. The analysis was performed according
groups of hypertensive rats receiving TPLE at to the procedural instructions included in the kit.
dosages of 50, 100, and 200 mg/kgBW. The rats NO concentrations were expressed in µmol.(2)

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Univ Med Vol. 32 No.1

The method used for determining Brawijaya University, Malang.

pulmonary MDA levels was developed at the
Pharmacology Laboratory, Faculty of Medicine, RESULTS
Brawijaya University, Malang, East Java. One
hundred milligrams of tissue was homogenized On the basis of DPPH radical scavenging
with 2 ml of phosphate buffer at pH 7.4. Then activity, TPLE was a strong antioxidant with an
50 ml of 0.1% Triton-X was added and the IC50 of 11.64%.
mixture was vortexed. Thereafter, 250 ml of 1 Figure 1 presents arterial NO levels in the
N HCL was added to the mixture, which was treatment groups. Post hoc testing found a
again vortexed. The mixture was heated to 1000 significant reduction in arterial NO in the non-
C for 30 minutes, then centrifuged at 3000 rpm TPLE hypertension group, as compared with the
for 10 minutes until a colloid precipitate formed. control group (p<0.001). To raise arterial NO,
The absorbance was read with a TPLE was administered daily for 6 weeks.
spectrophotometer at a wavelength (ë) of 532 Administration of TPLE at a dosage of 200 mg/
nm. The MDA concentration was expressed in kgBW was able to significantly increase arterial
ng/200 mg. NO, as compared with non-TPLE hypertensive
rats (p<0.001), although the increase did not
Data analysis attain the levels in control rats.
To examine the differenceas in arterial NO In Figure 2 are presented the pulmonary
and pulmonary MDA, the data were tested with MDA concentrations in the treatment groups.
ANOVA and a post hoc test. A value p<0.05 Post hoc testing found a significant increase in
was considered to indicate a significant pulmonary MDA concentration in rats with non-
difference. The statistical analysis was done TPLE hypertension as compared with control
using SPSS version 13. rats (p<0.05). Administration of TPLE at the
dosages of 100 and 200 mg/kgBW was able to
Ethical clearance significantly reduce MDA levels in comparison
This study was approved by the Committee with non-TPLE hypertensive rats, although not
on Research Ethics, Faculty of Medicine, attaining the levels in control rats.

Figure 1. NO concentration (µmol) in the treatment groups. Data are presented as mean ± standard deviation.
Dosage unit of TPLE is mg/kgBW; *significantly different in comparison with control group (p<0.05); #
significantly different in comparison with non-TPLE hypertension group (p<0.05)

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Athiroh, Sulistyowati Scurrula atropurpurea and hypertension

Figure 2. MDA concentration (ng/200 mg) in the treatment groups. Data are presented as mean ± standard
deviation. Dosage unit of TPLE is mg/kgBW; *Significantly different in comparison with control group
(p<0.05); # significantly different in comparison with the non-TPLE hypertension group (p<0.05)

DISCUSSION compared with the group of non-TPLE

hypertensive rats, but is not able to attain the
The present study demonstrates that in the NO levels in the control rats. The ability of
rat model of hypertension there is a significantly TPLE to modulate NO levels is due to its active
reduced arterial NO concentration and a constituents, among others flavonol glycosides
significantly increased pulmonary MDA (quercetin and rutin), monoterpene glucosides
concentration, in comparison with the control (icariside B), lignan glycosides (aviculin),
group. The reduction in arterial NO level flavans (catechin, epicatechin, epicatechin-3-
indicates the occurrence of endothelial 0-gallate, epigallocatechin-3-0-gallate,
dysfunction in hypertension. The increase in gallocatechin, and epigallocatechin). (6)
pulmonary MDA indicates an increase in Quercetin diffuses directly into endothelial cells
oxidative stress in hypertension. Endothelial and increases NO production. (13) Catechin
dysfunction and pulmonary oxidative stress in increases eNOS phosphorylation and NO
hypertension may be modulated by the bioavailability by inhibition of NADPH
administration of TPLE. oxidase. (14)
The reduced arterial NO in hypertension In the present study, pulmonary oxidative
may be due to decreased expression of stress increased significantly in comparison with
endothelial nitric oxide synthase (eNOS) or by the controls. This indicates pulmonary
decreased NO bioavailability.(11) The latter may involvement in the rat model of hypertension
be the result of a reaction between NO and induced by DOCA. In the vascular system, one
superoxide radicals to form peroxynitrites.(12) of the mechanisms for increases in reactive
Subsequently peroxynitrites may trigger the oxygen compounds is the high activity of
formation of hydroxyl radicals leading to lipid NADPH oxidase. (15) We suppose that this
peroxidation, which is marked by an increase in mechanism is also at work in the lungs.
MDA levels. Administration of TPLE at dosages of 100
Administration of crude TPLE at a dosage and 200 mg/KgBW is capable of significantly
of 200 mg/KgBW is able to significantly decreasing pulmonary MDA levels in
increase arterial NO concentration, as comparison with the group of non-TPLE

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Univ Med Vol. 32 No.1

hypertensive rats, but cannot attain the MDA and suggestions; 3). The umbrella research team
levels in the control rats. This indicates that at the Faculty of Mathematics and Natural
TPLE has antioxidant properties and inhibits Sciences (FMIPA) and the Faculty of Medicine,
pulmonary oxidative stress. Based on its DPPH Islamic University of Malang (UNISMA) for
radical scavenging activity, TPLE is a strong their cooperation.
antioxidant with an IC 50 value of 11.64%.
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