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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
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
Article history:
Received 28 September 2015
Accepted 15 November 2015
Available online 24 November 2015
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
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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
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
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
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.
This study had certified by Ethic commission of Faculty of Medicine, University of Brawijaya, Indonesia
with letter approval number: 369/ EC/ KEPK/06/2015.
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
ALT (U/l)
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
Liver function biomarkers (U/l) Ẋ ± SD
Total Protein
Liver Weight
124.80 ± 19.33
74.40 ± 13.93
3.35 ± 0.22
3.45 ± 0.43
6.80 ± 0.38
7.64 ± 1.23
136.20 ± 22.60
77.80 ± 19.09
3.35 ± 0.11
3.56 ± 0.60
6.91 ± 0.53
6.64 ± 0.85
149.00 ± 24.60
65.40 ± 11.01
3.40 ± 0.19
3.42 ± 0.41
6.82 ± 0.53
6.38 ± 0.79
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:
AST (u/L)
1. AST serum
Albumin (U/l)
3. Albumin serum level
Globuline (U/l)
4. Globulin serum level
Total Protein (U/l)
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].
7. MESA 1
8. MESA 2
9. MESA 3
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].
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.
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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BACKGROUND Hypertension is the most prevalent chronic disease and has an impact on one billion people. Production of superoxide radicals and endothelial dysfunction are involved in hypertension. Scurrula atropurpurea (BL.) Dans. is a tea plant parasite. This study aimed to evaluate the role of Scurrula atropurpurea (BL.) Dans. on nitric oxide (NO) as a marker of endothelial dysfunction and malondialdehyde (MDA) as a marker of oxidative stress in hypertensive rats. METHODS This study subjected rats to deoxycorticosterone acetate (DOCA)-induced hypertension. The experimental groups consisting of the control group and 3 hypertension groups receiving Scurulla tropurpurea extract at a dosage of 50; 100; and 200 mg/KgBW. Scavenging activity of Scurrula atropurpurea (BL.) extract was analyzed by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. The levels of arterial nitric oxide (NO) and pulmonary malondialdehyde (MDA) were analyzed by spectrophotometry. ANOVA and a post hoc test were applied to find the difference of arterial NO and pulmonary MDA levels between groups. RESULTS 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 significantly increased (p
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To know whether Scurrula atropurpurea is able to modulate total plasma nitrate/nitrite levels, decrease endothelial damage, and increase endothelial progenitor cells (EPCs) in hypertensive rats. The rats were divided in 5 groups: control (normotensive) group, Desoxy cortico sterone (DOCA)-salt hypertensive group, and three DOCA-salt hypertensive groups. All 5 groups received methanolic extract of S. atropurpurea (MESA) at a dosage of 50; 100; and 200 mg/KgBW. Serum nitric oxide (NO) was assayed by colorimetric. Circulating endothelial cells (CECs) and EPCs were assayed using flow cytometry. The administration of MESA100 and MESA200 elevated the total plasma nitrate/nitrite levels but cannot reach the level in control group. MESA100 and MESA200 also elevated the EPCs number compared with hypertensive group. The administration of MESA significantly (P< 0.05) decreased the CECs number compared to hypertensive groups. Methanolic extract of S. atropurpurea is able to modulate total plasma nitrate/nitrite levels and diminish endothelial damage via increasing EPCs.
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Mistletoes of the Loranthaceae and Viscaceae are hemiparasitic plants and their preparations in the form of injectable extracts, infusions, tinctures, fluid extracts or tea bags are widely used in various cultures in almost every continent to treat or manage various health problems including hypertension, diabetes mellitus, inflammatory conditions, irregular menstruations, menopause, epilepsy, arthritis, cancer, etc. The medicinal values of some species of Mistletoes (Loranthaceae) growing in the West African sub-region have been reviewed along with some considerations of their chemistries and local uses. These have been compared with Mistletoes (Loranthaceae and Viscaceae) growing elsewhere in Europe and Asia. This review has attempted to update our knowledge on the values of these hemi-parasites which belong to the genera - Globimetula, Phragmanthera, Agelanthus and Tapinanthus, and which have, for years, been seen as only devastating and notorious plants. They are also seen as epiphyting economic, ornamental and medicinal plants. The hemi-parasitic plants (Mistletoes) are not well understood as very little is known about their biology (taxonomy, host/plant relationship, ecology, toxicology, physiological characteristics, etc.) and chemistry (chemical constituents' profile). Some pharmacological studies carried out on the various crude alcoholic extracts and purified fractions have, however, revealed that mistletoes showed hypotensive, hypoglycaemic, antilipidaemic, anti-oxidative, anti-inflammatory, antimicrobial, etc. effects and were non-toxic in experimental animals at the doses used. The findings showed that mistletoes can be very useful as medicinal agents in ameliorating health problems such as diabetes mellitus, hypertension, arthritis, pain, cancer and a host of other ailments if properly studied and developed.
This study aimed to investigate whether methanolic extract of Scurrula atropurpurea (BL.) Dans. able to diminisihed oxidative stress in hypertensive rats. This study subjected rats to DOCA-induced hypertension. The experimental design: I. Sham group, II. Hypertensive group (administered by DOCA-salt at dosage of 10 mg/KgBW). III-V. The groups receiving Scurulla atropurpurea (BL.) Dans. methanolic extract at dosage of 50; 100; and 200 mg/KgBW. The levels of blood malondialdehyde (MDA) and superoxide dismutase (SOD) were measurenmet by spektrofotometry. There was significantly (P < 0.05) increased systolic blood pressure in hypertension rats compared to sham group. Compared to its hypertensive group, the administration of MESA50 significantly decreased systolic blood pressure, but not able to reach the level in sham group. The SOD level were significantly (P < 0.05) decreased in hypertension rats compared to sham group. The administration of MESA100 elevated the SOD levels to reach level in sham group. The SOD levels in MESA100 and MESA200 were higher significantly (P < 0.05) compared to hypertensive group. There were significantly (P < 0.05) increased MDA levels in hypertension rats compared to sham group. The administration of MESA200 significantly (P < 0.05) decreased the MDA levels compared to hypertensive groups. In conclusion, Scurulla atropurpurea (BL.) Dans. methanolic extract able to decreased sistolic blood pressure, modulates SOD, and diminished oxidative stress (MDA), and in DOCA-salt hypertensive rats. Copyright