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Advances in Natural and Applied Sciences, 8(4) April 2014, Pages: 233-236
AENSI Journals
Advances in Natural and Applied Sciences
ISSN:1995-0772 EISSN: 1998-1090
Journal home page: www.aensiweb.com/anas/index.html
Anti-hyperglycemic Activity Studies on Leaves and Stems of Areca catechu L.
(Arecaceae)
Shirin Akhter, Maruf Hassan, Shahnaz Rahman, Afifa Sultana, Shejuty Shahreen, Joyonta Banik, Rownak
Jahan, Mohammed Rahmatullah
Department of Biotechnology and Genetic Engineering, University of Development Alternative, Dhanmondi, Dhaka-1205, Bangladesh
ARTICLE INFO
Article history:
Received 25 January 2014
Received in revised form 12
March 2014
Accepted 14 April 2014
Available online 5 May 2014
Keywords:
Areca catechu; Hypoglycemic activity;
Serum glucose level; Glibenclamide.
ABSTRACT
Areca catechu L. (Arecaceae) leaves and stems are used in folk medicinal system of
Bangladesh for treating intestinal pain, ulcer, dental disease, diarrhea, gout, and
constipation, but so far no scientific study has been done with regard to their antihyperglycemic effects. This study was undertaken to evaluate the possible glucose
tolerance efficacy of methanolic extracts of Areca catechu leaves and stems using
glucose-induced hyperglycemic mice. Different doses of the extracts were administered
an hour before glucose administration and blood glucose levels measured two hours
following glucose administration (p.o.) using glucose oxidase method. The statistical
data indicated significant and dose-dependent oral hypoglycemic activity on glucoseloaded mice at all doses of the extracts tested. Maximum anti-hyperglycemic activity
was shown at 400 mg leaf as well as stem extract per kg body weight, which was
comparable to that of glibenclamide (a standard drug for comparing anti-hyperglycemic
activity, administered at a dose of 10 mg/kg body weight). Dose for dose, the leaf
extract showed more anti-hyperglycemic effects than the stem extract.
© 2014 AENSI Publisher All rights reserved.
To Cite This Article: Shirin Akhter, Maruf Hassan, Shahnaz Rahman, Afifa Sultana, Shejuty Shahreen, Joyonta Banik, Rownak Jahan,
Mohammed Rahmatullah., Anti-hyperglycemic Activity Studies on Leaves and Stems of Areca catechu L. (Arecaceae). Adv. in Nat. Appl.
Sci., 8(4): 233-236, 2014
INTRODUCTION
Areca catechu L. (Arecaceae) (local name: supari) is more commonly known as the areca palm or betel nut
palm because its fruit is chewed often with betel leaf. The palm tree can grow up to 20 meters tall, and is
cultivated throughout the tropical parts of the Indian sub-continent, including Bangladesh (Satyavatia et al.,
1976). The fruits of the palm (nuts) are known to be in use for over two thousand years (Bradley, 1979) and are
chewed either by themselves for breath freshening effect, digestive aid, expelling intestinal worms (Cawte,
1985), or as in Bangladesh, with betel leaf, lime and other ingredients as a digestive aid and stimulant effect.
Betel nut with betel leaf chewing is possibly the biggest addiction in Bangladesh, more so than tobacco.
Chewing of the nut reportedly caused a depressive effect on the heart and hypotension (Kapoor, 1990; Lin et al.,
2002).
A number of alkaloids have been reported to be present in the nut. These include arecoline, arecaidine,
guvacoline, guvacine, arecolidine, and choline (Chu, 2001; Farnsworth, 1976). Presence of phenolic compounds
like hydroxychavicol and safrole, and phytochemicals like tannins, gallic acid, catechin, -sitosterol, gum and
amino acids have been reported in betel nut (Wang et al., 1997; Duke, 1992).
The anti-depressant effects of ethanol extract of betel nuts have been reported (Dar and Khatoon, 1997).
Cholinomimetic and acetylcholinesterase inhibitory constituents have also been reported in betel nut (Gilani et
al., 2004). Arecoline, an alkaloid present in betel nut reportedly showed hypoglycemic activities (Chempakam,
1993). The compound also reportedly enhanced uptake of 2-deoxyglucose dose-dependently in L6 myotubes
and caused increased expression of glucose transporter type 4 (GLUT4) and phosphoinositide 3-kinase (PI3K)
genes suggesting that the compound can replace commercially available anti-diabetic drugs (Prabhakar and
Doble, 2010). On the other hand, it has been reported that arecoline significantly attenuated in 3T3-L1
adipocytes insulin-induced uptake of glucose (Hsu et al., 2010). Clearly, more studies are necessary for accurate
evaluation of betel nut extract or isolated constituents like arecoline from nut for any possible anti-diabetic
effects.
Corresponding Author: Professor Dr. Mohammed Rahmatullah, Pro-Vice Chancellor and Dean, Faculty of Life Sciences,
University of Development Alternative, House No. 78, Road No. 11A (new), Dhanmondi, Dhaka1205, Bangladesh
Tele: +88-01715032621
Fax: +88-02-815739
E-mail: [email protected]
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Dr. Mohammed Rahmatullah et al,2014
Advances in Natural and Applied Sciences, 8(4) April 2014, Pages: 233-236
Contrary to betel nut and some of its constituents, there is an absolute dearth of pharmacological activity
studies on effect of betel leaf and stem extracts. The leaves and stems are used in folk medicine of Bangladesh
for the treatment of intestinal pain, ulcer, dental disease, diarrhea, gout, and constipation, but so far no scientific
study has been done with regard to their anti-hyperglycemic effects. It was the objective of the present study to
evaluate the anti-hyperglycemic activity of leaves and stems of the plant in a rodent model.
MATERIALS AND METHODS
Collection of plant material:
The leaves and stems of Areca catechu were collected in December 2009 from Gazipur district,
Bangladesh. Identification of the leaves and stems was done at the Bangladesh National Herbarium, Mirpur,
Dhaka (Accession No. 35,030) and sample specimens have been kept over there.
Preparation of the test samples:
The dried leaves and stems of Acacia catechu were separately air-dried in the shade and pulverized into a
fine powder. The powder was then mixed with methanol at a ratio of 1:3 (w/v). After 24 hours, the mixtures
were filtered; filtrate was collected and the residue was again mixed with fresh methanol at a ratio of 1:2 (w/v)
for 24 hours. After filtration, filtrates were combined and evaporated to dryness (approximate yields were 11.2%
for leaves and 4.1% for stems) using rotary evaporator. 1% Tween 80 in water was used to suspend the extracts
prior to administration.
Preliminary phytochemical screening:
Preliminary phytochemical screening (Kokate, 1994; Harborne, 1998) revealed the presence of presence of
phenolic compounds, alkaloids, glycosides, and tannins.
Animals:
Swiss albino mice (male), weighing 25-30g bred in the animal house of ICDDR,B (International Centre for
Diarrheal Disease and Research, Bangladesh) were used for the present experiments. All animals were
acclimatized one week prior to the experiments. The animals were housed under standard laboratory conditions
(relative humidity 55-65%, room temperature 25.0 ± 20C, and 12 hrs light-dark cycles). The animals were fed
with standard diet from ICDDR,B and had free access to water. The study was approved by the Institutional
Animal Ethical Committee of the University of Development Alternative, Dhaka, Bangladesh.
Anti-hyperglycemic activity test:
Anti-hyperglycemic activities of the extracts were studied through the glucose tolerance test method.
Glucose tolerance test was performed following the procedure described by Joy and Kuttan (1999) with minor
modifications. Briefly, fasted mice were divided into nine groups of six mice each. Each group received a
particular treatment like group-I served as control and received vehicle (1% Tween 80 in water, 10 ml.kg -1 body
weight), while group-II received standard drug (glibenclamide, 10 mg.kg-1 body weight). Groups III-VI received
stem extract at four different doses of 50, 100, 200 and 400 mg extract.kg -1 body weight, respectively. Groups
VII-IX received leaf extract at doses of 100, 200 and 400 mg extract.kg-1 body weight, respectively. Each mouse
was weighed properly and the doses of the test samples, standard drug, and control materials were adjusted
accordingly. Test samples, control, and glibenclamide were given orally. After one hour, all mice were orally
treated with 2 g.kg-1 of glucose. Blood samples were collected two hours after glucose administration. Serum
was separated and blood glucose levels were measured immediately by glucose oxidase method (Venkatesh et
al., 2004).
Statistical analysis for anti-hyperglycemic activity:
Experimental results are expressed as mean ± SEM. For statistical comparison, Independent Sample t-test
was carried out. A p value < 0.05 in all cases was considered to be statistically significant.
Acute toxicity study:
The study was carried out as has been previously described (Ganapaty et al., 2002) with minor
modifications. Selected animals were divided into nine groups of six animals each. The control group received
1% Tween 80 in normal saline (2 ml.kg-1 body weight). The other groups received respectively, 100, 200, 300,
600, 800, 1000, and 2000 mg leaf or stem methanolic extract.kg -1 body weight. Animals were monitored closely
after dosing for the next 8 hours for any behavioral changes and were kept under observation up to 14 days to
find out if there is any mortality.
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Dr. Mohammed Rahmatullah et al,2014
Advances in Natural and Applied Sciences, 8(4) April 2014, Pages: 233-236
RESULTS AND DISCUSSION
Acute toxicity study:
Mice did not show any mortality with any of the extracts at tested doses till the end of 14 days of
observation.
Table 1: Effect of methanol extract of Areca catechu (leaf and stem) on serum glucose level in hyperglycemic mice.
Treatment
Dose
Serum glucose level (mg.dl-1)
% inhibition
Group I (Control, vehicle)
1% Tween 80 in water
84.2 ± 9.1
(10 ml.kg-1 body weight)
-1
Group II (glibenclamide)
10 mg.kg body weight
24.1 ± 2.5*
71.4
Group III (stem extract)
50 mg.kg-1 body weight
48.7 ± 6.6*
42.2
Group IV (stem extract)
100 mg.kg-1 body weight
36.4 ± 2.8*
56.8
Group V (stem extract)
200 mg.kg-1 body weight
34.2 ± 3.3*
59.4
Group VI (stem extract)
400 mg.kg-1 body weight
31.1 ± 3.7*
63.1
-1
Group VII (leaf extract)
100 mg.kg body weight
34.7 ± 2.8*
58.8
Group VIII (leaf extract)
200 mg.kg-1 body weight
32.9 ± 3.5*
60.9
Group IX (leaf extract)
400 mg.kg-1 body weight
28.9 ± 2.5*
65.7
Extracts and drug were given orally one hour before glucose administration and serum glucose level was measured two hours after glucose
administration. Values are given as Mean ± S.E.M. from six mice in each group. * P < 0.05 is significant compared to hyperglycemic control
animals.
Anti-hyperglycemic effect:
The results obtained from the present study indicate that the methanol extract of the leaves and stems of
Areca catechu lowered serum glucose levels dose-dependently and significantly when compared to control
(group-I) at all the doses examined. Dose for dose, the anti-hyperglycemic activity was more pronounced with
methanolic leaf extract of the plant than the stem extract. Maximum hypoglycemic activity of methanol extract
of Areca catechu leaves in glucose-induced hyperglycemic mice was observed with a 400 mg.kg-1 dose (65.7%
inhibition), while the standard drug, glibenclamide produced 71.4 % inhibitory activity at 10 mg.kg-1 dose
(Table 1) under the experimental conditions of the present study. The present preliminary experimental results
indicated that leaves and stems of Areca catechu exhibited significant blood glucose lowering property in
glucose-induced hyperglycemic mice. A known hypoglycemic agent – arecoline is reportedly present in nuts of
the plant (Chempakam, 1993). Whether arecoline is present in leaves and stems of the plant is currently under
investigation. This, however, dose not preclude the presence of other phytochemical components in leaves and
stems of Areca catechu, which was responsible for the observed anti-hyperglycemic effect. The mechanism
underlying the glucose lowering efficacy of Areca catechu is yet to be investigated and is currently going on in
our laboratory. However, it is speculated that the glucose lowering activity of the extract may either be through
potentiating insulin secretion by the pancreas or through increasing the uptake of glucose.
It is to be noted that antihyperglycemic activity of leaf extract of A. catechu has been reported in
streptozotocin-induced diabetic rats (Mondal et al., 2012). The beneficial role of nut extract in alloxan diabetic
rats has also been reported (Kavitha et al., 2013). Ethanolic extract of leaf has been shown to cause significant
lowering of blood glucose in hyperglycemic Sprague Dawley rats (Wankhade and Nandi, 2013). Thus the
results obtained in the present study are consistent with previous studies and suggest that constituents from A.
catechu can prove to be valuable glucose lowering agents in diabetic patients.
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