Evaluation of Methanolic Extract of Scurrula Atropurpurea (Bl.) Dans Sub-Chronic Exposure On Wistar Rat Liver

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Advances in Environmental Biology, 9(23) October 2015, Pages: 245-250
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Advances in Environmental Biology
ISSN-1995-0756 EISSN-1998-1066
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Corresponding Author: N. Athiroh, Department of Biology, Faculty of Mathematic and Natural Sciences, Islamic
University of Malang, East Java Indonesia
E-mail: nur_athiroh_mlg@yahoo.co.id
Evaluation of Methanolic Extract of Scurrula Atropurpurea (Bl.) Dans Sub-Chronic
Exposure On Wistar Rat Liver
1Nour Athiroh and 2Erna Sulistyowati
1Department of Biology, Faculty of Mathematic and Natural Sciences, Islamic University of Malang, East Java Indonesia
2Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, R.O.C
A RT I C LE I NF O
A B ST R AC T
Article history:
Received 28 September 2015
Accepted 15 November 2015
Available online 24 November 2015
Keywords:
liver protection, Scurrula
atropurpurea (Bl.) Dans
Background: This study aimed to analyze sub-chronically exposure of methanolic
extract of Scurrula atropurpurea (Bl.) Dans (MESA) on rat liver function such as AST
(aspartate transferase), ALT (alanine transaminase), level of serum albumin, level of
serum globuline level of total serum protein, and histopathology of liver. Objectives:
Male Wistar rats were randomly divided into four groups: control, MESA I (250
mg/BW), MESA II (500 mg/BW), MESA III (1000 mg/BW). Methanolic extract was
given once daily per oral for 28 days. Body weight were measured twice weekly. Data
were analyzed by one way ANOVA test and continued by Post Hoc test on SPSS 21
software. Results: No effect were found on orally exposure of MESA for 28 days
compared to the control, on rat level of serum AST, serum ALT, level of serum
albumin, level of serum globuline and level of total serum protein (p > 0.05). That
besides no abnormality in liver histopathology in treatment. Conclusion: This study
concluded sub-chronic exposure of MESA did not interfered rat liver function. It would
be a recommendation for next clinical study
© 2015 AENSI Publisher All rights reserved.
To Cite This Article: Nour Athiroh, Erna Sulistyowati, Evaluation Of Methanolic Extract Of Scurrula Atropurpurea (Bl.) Dans Sub-
Chronic Exposure On Wistar Rat Liver. Adv. Environ. Biol., 9(23), 245-250, 2015
INTRODUCTION
Scurrula atropurpurea (Bl.) Dans is a a half parasitic plant cause has chlorophyll and remains able to
assimilate itself, suck the water as well as organic and inorganic tea from host plant. Botanical classification of
Scurrula atropurpurea (Bl.) Dans is class Dicotyledonae, Santales order, Loranthaceae family. Scurrula has
several species; Scurrula (S) oortiana, S. atropurpurea, S. junghuni, and S.parastica. Scurrula atropurpurea
extract contains 16 bioactive materials comprising of six fatty acid compounds, two santin, two monoterpenes
glycosides of the flavonol glycosides, one lignan glycosides, and four flavones [1,2]. Several types of group of
Loranthaceae family potentially have antihypertensive. Many studies reported possibility of parasitic plant on
tea crop had antihypertensive role, one is Viscum album. Administration of Viscum album for 3-5 weeks abled
to reduce blood pressure on hypertensive patient [3,2].
Other types of parasitic plant on tea, Scurulla oortiana and parasitic on guava roses (Macrosolen javanus)
had capability to decrease contraction of separated arterie on rat in vitro study [2]. Previous study, S.
atropurpurea showed its ability on lowering blood pressure in DOCA-salt hypertensive rats, through
improvement of endothelial dysfunction and oxidative stress [4,5,6]. The activity of active ingredients in S.
atropurpurea are antiantioxidant which inhibit oxidative damage caused by free radicals.
The leaves and stems of these plants contain alkaloids, flavonoids, glycosides, triterpenes, saponins, and
tannins that act as antioxidants. Potential flavonoids as antioxidants can reduce the activity hydroxy radical,
superoxide anion and peroxide radicals fat [1,2].
Based on these studies, we evaluate sub-chronical administration of S. atropurpurea on liver function to
show its safety. This study generated to standardize it as an antihypertensive medicinal preparation. For that
reason we aimed to analyse level of liver enzymes; ALT, AST, albumin, globulin, and total protein in Wistar
rats that were given S. atropurpurea for 28 days.

246 N. Athiroh et al, 2015
Advances in Environmental Biology, 9(23) October 2015, Pages: 245-250
Methodology:
Preparation of tea parasite crude extract:
Preparation of tea parasite crude extract Scurrula atropurpurea was determined biologically at the
Laboratorium of Material Medica Batu East Java. MESA was obtained through several steps. The leaves were
washed, dried in an oven at 40-600C, then ground into a powder. A 100 mg portion of this powder was steeped
in methanol in a 1 L Erlenmeyer flask. The mixture was shaken for 30 minutes to distribute the powder
homogenously in the methanol. To collect the precipitate, the mixture was left to stand overnight. The upper
layer know and supernatant, being a mixture of methanol and the active constituents, was subjected to
evaporation. The extract was then labeled [4,5,6,7].
Animal studies:
Male Wistar rats, weight 200-300 gram, were acclimatized for 7 days, grouped randomly, and put on cages
according to weight evenly (weight variation less than 20% of average weight. The rats were fasted for 14-18
hours before it had given treatment. Drinking water were provided. After fasted, Rats were weighed and given
MESA by oral gavage. Once treated, the feed should be given back after 3-4 hours. The dosage of MESA
defined on 500 mg/kg/BW as previously described [4,5,6,8]. MESA administered daily for 28 days with volume
of dilution was 1 ml/100g BW. Body weight measured twice a week. At day 29th rats were sacrified and blood
was drawn to observe levels of AST, ALT, albumin, globulin and total protein serum level.
Blood Samples:
The blood samples were collected, left to clot in clean, dry tubes, and centrifuged at 3000 rpm (600 g) for
10 minutes using Heraeus Labofuge 400R, Kendro Laboratory Products GmbH, Germany, to separate sera. The
sera were kept in a deep freezer at −20°C until biochemical markers were analyzed within one week.
Liver Biomarkers:
All biochemical measurements were determined in serum according to the details given in the kit’s
instructions and performed by using a Shimadzu UV-VIS Recording 2401 PC (Japan). The activities of cellular
enzymes such as AST and ALT were determined according to the methods of Reitman and Frankel (9), while the
concentrations of albumin, globulin and total protein were determined according to the methods of Lowry et al.,
[10, 11, 12, 13, 14] respectively.
Histopathology of Liver Tissues:
Adequate fixation is crucial to the success of histopathological evaluation. Approximately twenty times the
volume of 10% neutral buffered formalin (NBF) relative to the amount of tissue to be fixed should be used.
Tissue samples should be less than 5mm thick to ensure thorough fixation. After tissue specimens are fixed in
10% neutral buffered formalin, they are dehydrated in graded alcohols, embedded in paraffin, sectioned at 3-5
µm, stained with hematoxylin and eosin (H&E), and cover-slipped for standard light microscopic
histopathological interpretation by the pathologist[15]. Histological slides were photographed under Olympus
photomicroscope.
Statistical analysis:
Statistical analysis was done using SPSS 17 for windows and the values were expressed as mean ± S.D. The
statistical significance of differences between the means was analyzed using one-way analysis of variance
(ANOVA) followed by Duncan’s test for comparison between different treatment groups. Statistical
significance was set at p<0.05.
Ethics:
This study had certified by Ethic commission of Faculty of Medicine, University of Brawijaya, Indonesia
with letter approval number: 369/ EC/ KEPK/06/2015.
RESULTS AND DISCUSSION
Signs of Toxicity:
No mortality occurred during the study period. We observed no symptoms and no signs of toxicity such as
hyperirritability on rats. In addition, there was no effect on food and water consumptions in all groups (in
tabulated data).
Liver Biomarkers:
The results demonstrating relatively normal on level of liver biomarkers. There were unsignificant increases
in AST and ALT in all groups (Table 1). AST serum level raised in each group (Figure 1) and also level of ALT

247 N. Athiroh et al, 2015
Advances in Environmental Biology, 9(23) October 2015, Pages: 245-250
A
A
A
A
0
50
100
Control
MESA I
MESA II
MESA III
ALT (U/l)
Groups
2. ALT serum level
(Figure 1). However, albumin, globulin and total protein levels were normal in all rats (Figure 1).
Supplementation of MESA 250, 500 and 1000 mg/kgBW daily for 28 days showed no changes of compare to
the control groups.
Table 1: Liver function biomarkers and liver weight in treatment group
Group
Liver function biomarkers (U/l) Ẋ ± SD
AST
ALT
Albumin
Globulin
Total Protein
Liver Weight
(gr)
Control
124.80 ± 19.33
74.40 ± 13.93
3.35 ± 0.22
3.45 ± 0.43
6.80 ± 0.38
7.64 ± 1.23
MESA I
136.20 ± 22.60
77.80 ± 19.09
3.35 ± 0.11
3.56 ± 0.60
6.91 ± 0.53
6.64 ± 0.85
MESA II
149.00 ± 24.60
65.40 ± 11.01
3.40 ± 0.19
3.42 ± 0.41
6.82 ± 0.53
6.38 ± 0.79
MESA III
122.80 ± 7.46
67.60 ± 16.71
3.55 ± 0.13
3.21 ± 0.35
6.76 ± 0.25
7 ± 1.02
Fig. 1: Serum exposed to MESA for 28 days. The values represented are the means ± S.D. A = means having
the same letters are not significantly different from each other, 𝑃 ≥ 0.05. Groups: control, MESA 1 =
250 mg/kg BW. MESA 2 = 500 mg/kg BW and MESA 3 = 1000 mg/kg BW
Histopathology of Liver:
A
A
A
A
0
50
100
150
200
Control
MESA I
MESA
II
MESA
III
AST (u/L)
Groups
1. AST serum
A
A
A
A
3
3.2
3.4
3.6
3.8
Control
MESA I
MESA
II
MESA
III
Albumin (U/l)
Groups
3. Albumin serum level
A
A
A
A
2.8
3
3.2
3.4
3.6
3.8
Control
MESA I
MESA
II
MESA
III
Globuline (U/l)
Groups
4. Globulin serum level
A
A
A
A
6.6
6.7
6.8
6.9
7
Control
MESA I
MESA II
MESA
III
Total Protein (U/l)
Groups
5. Total Protein serum level

248 N. Athiroh et al, 2015
Advances in Environmental Biology, 9(23) October 2015, Pages: 245-250
Fig. 2 Histopathology of rat liver exposed to MESA for 28 days. There is no abnormality effect. H.E. X 100. a =
A portal triad ; b = vena centralis ; c = nucleus cell
Results of the present study demonstrate that subchronic administration of MESA did not produce signify
toxicity. Providing MESA cause significant increase in body weight gain in rats. There is no effect on feed and
water intake in treated rats. In addition, there are no differences in MESA groups compared to control on liver
function. Other study, Amin, et. al showed ethanolic extract of Phyllanthus niruri and Melastoma
malabathricum able to improve liver function due to increasing immune system[16].
Liver function biomarkers are a helpful screening tool, which are an effective modality to detect hepatic
dysfunction. Our results showed that MESA have no caused in liver enzymes, that is, AST and ALT in Wistar
rats. In fact, aminotransferases are intracellular enzymes, most frequently utilized, and specific indicators of
hepatocellular necrosis. AST and ALT catalyze the transfer of the amino acids of aspartate and alanine,
respectively, to the keto group of ketoglutaric acid.
In the present study, oral administration of MESA to Wistar rats disable cause changes in serum total
protein, albumin and globulin levels. This study proved MESA has no effects in causing liver dysfunctions and
disturbance in the biosynthesis of protein. Several tea parasite noted that several studies showed Loranthaceae
family in various feeding experiments have no toxicity. those tests carried out suggest that the crude mistletoe
leaf extracts of Loranthaceae family were non-toxic to laboratory animals [17].
Other potential role of MESA is protecting from microbes. It competent to kill bacteria. Scurrula
atropurpurea inhibit 50% concentration of Enterobacter sakazakii effectively [18]. Other studies tested it
suppressed Bacillus subtulis, Klebsella pneumaneae, Vibrio cholerae, and Esherichia coli. Scurrula
atropurpurea, Macrosolon cochichinensis and Viscum album are hemiparasites found in the forests of South
West Bengal, showed antimicrobial activities against four bacterial strains (Bacillus subtilis, Klebsella
pneumoniae, Vibrio cholerae and Escherichia coli). With regard to their natural distribution in the forests of
southern parts of west Bengal it was observed that the Scurrula atropurpurea and Viscum album are now very
scant and difficult to locate many times in the field [19]. Other hand that some chemical entities transferred
from host Dendrophthoe falcata and Mangifera indica to the parasite Scurrula parasiica are responsible for its
effect on blood sugar level [20].
a
b
c
6. CONTROL
ControlCO
NTROL
7. MESA 1
a
b
c
8. MESA 2
c
b
a
9. MESA 3
a
b
c

249 N. Athiroh et al, 2015
Advances in Environmental Biology, 9(23) October 2015, Pages: 245-250
The figure 2 showed that histopathology of liver in rat administered with MESA sub chronic 28 day. The
result, no e No abnormality was found in the histopathology of the liver, kidney, heart, and lung in the
experimental group of rats following same dose of two compounds isolated from Loranthus globosus Roxb
when compared with control group. This preliminary study suggest that isolated compounds may used safely for
clinical trial [21].
Conclusion:
The result, no e No abnormality was found in the histopathology of the liver, kidney, heart, and lung in the
experimental group of rats following same dose of two compounds isolated from Loranthus globosus Roxb
when compared with control group. This preliminary study suggest that isolated compounds may used safely for
clinical trial.
Conflicts of Interest:
The authors declare that there are no conflicts of interests regarding the publication of this article.
ACKNOWLEDGMENT
The authors thank to Direktorat Jenderal Pendidikan Tinggi, Kementerian Pendidikan dan Kebudayaan, for
funding on penelitian Strategis Nasional letter number : 018/SP2H/P/K7/KM/2015, on April 2nd 2015.
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