Antihyperglycemic activity studies with methanol extract of Madhuca indica j.f. gmel. leaves and Paederia foetida L. stems in mice

Abstract

Antihyperglycemic activity studies were conducted by oral glucose tolerance tests in glucose-loaded Swiss albino mice with crude methanol extract of leaves of Madhuca indica and stems of Paederia foetida. The leaf extract of Madhuca indica, when administered to mice at dose levels of 50, 100, 250, and 500 mg/kg body weight demonstrated dose-dependent and significant reductions in serum glucose levels at the three higher doses. Serum glucose levels were reduced by 22.2, 25.8, and 36.3%, respectively, at doses of 100, 250, and 500 mg extract/kg body weight. In comparison, the standard antihyperglycemic drug, glibenclamide, reduced serum glucose levels by 35.9%, which is approximately equivalent to that obtained with the highest dose of Madhuca indica leaf extract. Crude methanol extracts of stems of Paederia foetida also exhibited significant and dose-dependent reductions in serum glucose levels when administered to glucose-loaded mice. The extract at the four different doses of 50, 100, 250, and 500 mg/kg body weight, reduced serum glucose levels by 7.7, 25.3, 31.0, and 31.2%, respectively. Overall, the results demonstrate significant antihyperglycemic activities of leaves of Madhuca indica and stems of Paederia foetida.

Full text

Advances in Natural and Applied Sciences, 5(2): 122-126, 2011
ISSN 1995-0772
This is a refereed journal and all articles are professionally screened and reviewed
ORIGINAL ARTICLE
122
Corresponding Author: Mohammed Rahmatullah, Faculty of Life Sciences University of Development Alternative
House No. 78, Road No. 11A (new) Dhanmondi, Dhaka-1205 Bangladesh.
Tele: +88-01715032621; Fax: +88-02-815739
E-mail: rahamatm@hotmail.com
Antihyperglycemic Activity Studies with Methanol Extract of Madhuca Indica J.F. Gmel.
Leaves and Paederia Foetida L. Stems in Mice
Salehin Khan, Dilara Zahan, Rajib Das, Dilruba Nasrin, Shamima Ahsan, Rasheda Ahmed,
A.F.M. Nazmus Sadat, A.B.M. Anwarul Bashar, Nusratun Nahar, Mohammed Rahmatullah
Faculty of Life Sciences, University of Development Alternative, Dhanmondi, Dhaka-1205, Bangladesh.
Salehin Khan, Dilara Zahan, Rajib Das, Dilruba Nasrin, Shamima Ahsan, Rasheda Ahmed, A.F.M.
Nazmus Sadat, A.B.M. Anwarul Bashar, Nusratun Nahar, Mohammed Rahmatullah: Antihyperglycemic
Activity Studies with Methanol Extract of Madhuca Indica J.F. Gmel. Leaves and Paederia Foetida
L. Stems in Mice
ABSTRACT
Antihyperglycemic activity studies were conducted by oral glucose tolerance tests in glucose-loaded Swiss
albino mice with crude methanol extract of leaves of Madhuca indica and stems of Paederia foetida. The leaf
extract of Madhuca indica, when administered to mice at dose levels of 50, 100, 250, and 500 mg/kg body
weight demonstrated dose-dependent and significant reductions in serum glucose levels at the three higher
doses. Serum glucose levels were reduced by 22.2, 25.8, and 36.3%, respectively, at doses of 100, 250, and
500 mg extract/kg body weight. In comparison, the standard antihyperglycemic drug, glibenclamide, reduced
serum glucose levels by 35.9%, which is approximately equivalent to that obtained with the highest dose of
Madhuca indica leaf extract. Crude methanol extracts of stems of Paederia foetida also exhibited significant
and dose-dependent reductions in serum glucose levels when administered to glucose-loaded mice. The extract
at the four different doses of 50, 100, 250, and 500 mg/kg body weight, reduced serum glucose levels by 7.7,
25.3, 31.0, and 31.2%, respectively. Overall, the results demonstrate significant antihyperglycemic activities
of leaves of Madhuca indica and stems of Paederia foetida.
Key words: Madhuca indica, Paederia foetida, Sapotaceae, Rubiaceae, antihyperglycemic
Introduction
Madhuca indica J.F. Gmel. (Family: Sapotaceae, local name: mohua, English name: moa tree) is commonly
observed in various parts of the Indian sub-continent, including Bangladesh. The tree attains a height of about
20 meters, and possesses evergreen or semi-evergreen foliage. In the folk medicinal system of Bangladesh,
various parts of the tree are used, namely whole young plants, leaves, stems, barks, roots, fruits, flowers, and
seeds. The different ailments treated with these parts include tuberculosis, rheumatoid arthritis, cholera,
paralysis, snake-bite, debility, tonsillitis, influenza, piles, arthritic pain, helminthiasis, low semen count,
headache, flatulency, and infections, besides being used as a blood purifier and as an antidote to poison. Two
protobassic glycosides, namely madhucosides A and B have been isolated from the bark of this tree (Pawar
and Bhutani, 2004). The two compounds showed significant inhibitory effects on both superoxide release from
polymorphonuclear cells, and hypochlorous acid generation from neutrophils.
Paederia foetida L. (Family: Rubiaceae, local name: gondho vadali, English name: skunk vine) is a
climbing plant. It can be found in Bangladesh, parts of India, China and Japan, southern Bhutan, Cambodia,
Taiwan, Indonesia, Laos, Malaysia, Myanmar, Nepal, the Philippines, Singapore, South Korea, Thailand, and
Vietnam. Whole plants, leaves, and fruits are used in folk medicines of Bangladesh for treatment of ailments

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Adv. in Nat. Appl. Sci., 5(2): 122-126, 2011
like hepatic disorders, rheumatoid arthritis, constipation, diabetes, coughs, asthma, itches, wounds, stomachache,
diarrhea, dysentery, pain, typhoid, pneumonia, toothache, cancer, flatulency, body ache, and bone fractures.
Antioxidant activity has been reported for fresh and dried plant extracts (Osman et al., 2009). Antidiarrheal
activity has been reported for 90% ethanolic extract of the plant in castor oil and magnesium sulfate-induced
diarrheal models in mice (Afroz et al., 2006). There have also been reports of the anti-inflammatory effects
of plant or plant part extracts (De et al., 1994; Srivastava et al., 1973). The plant reportedly also inhibited
gastrointestinal helminthes in bovines (Roychoudhury et al., 1970).
Folk medicinal system is one form of traditional system practiced by folk medicinal practitioners or
Kavirajes in Bangladesh. Kavirajes, in fact, form the first tier of primary health care system in the country.
They rely almost exclusively on simple formulations of medicinal plants for treatment of various ailments, and
have quite extensive knowledge on the medicinal properties of different plant species. In our laboratory, we
had been studying plants used in the folk medicinal system of Bangladesh, particularly as to their
antihyperglycemic and antinociceptive effects (Anwar et al., 2010; Jahan et al., 2010; Khan et al., 2010;
Mannan et al., 2010; Rahman et al., 2010; Rahman et al., 2011; Shoha et al., 2011; Sutradhar et al., 2011).
The objective of the present study was to evaluate the antihyperglycemic potential of Madhuca indica leaves
and Paederia foetida stems in oral glucose tolerance tests in glucose-loaded mice.
Materials and methods
Plant Material and Extraction
The leaves of Madhuca indica were collected from Comilla district, Bangladesh in August, 2009. The
plant was taxonomically identified by the Bangladesh National Herbarium at Dhaka. The leaves of Madhuca
indica were air-dried in the shade for 120 hours, grounded into a fine powder, and were extracted with
methanol at a ratio of 1:5 (w/v). After 24 hrs, the mixture was filtered; filtrate was collected and the residue
was again extracted with methanol at a ratio of 1:3 (w/v) for 24 hrs. Filtrates were combined and evaporated
to dryness. The initial weight of dried leaf powder used for extraction was 100g; the final weight of the extract
was 7.3g. The stems of Paederia foetida were collected from Dhaka during June, 2010. Stems were sliced into
small pieces and air-dried in the shade for 120 hours. Methanol extraction of stems was done as described
before for leaves of Madhuca indica. The initial weight of dried stem powder used for extraction was 100g;
the final weight of the extract was 6.8g.
Chemicals and Drugs:
Glacial acetic acid was obtained from Sigma Chemicals, USA; aspirin, glibenclamide and glucose were
obtained from Square Pharmaceuticals Ltd., Bangladesh. All other chemicals were of analytical grade.
Animals
In the present study, Swiss albino mice (male), which weighed between 19-22g were used. The animals
were obtained from International Centre for Diarrheal Disease Research, Bangladesh (ICDDR,B). All animals
were kept under ambient temperature with 12h light followed by a 12h dark cycle. The animals were
acclimatized for three days prior to actual experiments. The study was conducted following approval by the
Institutional Animal Ethical Committee of University of Development Alternative, Dhaka, Bangladesh.
Anti-hyperglycemic Activity
Glucose tolerance property of methanol extract of Madhuca indica leaves and Paederia foetida stems was
determined as per the procedure previously described by Joy and Kuttan (1999) with minor modifications. In
brief, for Madhuca indica leaves, fasted mice were grouped into six groups of six mice each. The various
groups received different treatments like Group-I received vehicle (1% Tween 80 in water, 10 ml/kg body
weight) and served as control, group-II received standard drug (glibenclamide, 10 mg/kg body weight) and the
other four groups (III-VI) received the methanol extract of Madhuca indica leaves at four different doses of
50, 100, 250 and 500 mg/kg body weight. Each mouse was weighed and doses adjusted accordingly prior to
administration of vehicle, standard drug, and test samples. All substances were orally administered. Following
a period of one hour, all mice were orally administered 2 g glucose/kg of body weight. Blood samples were
collected two hours after glucose administration through puncturing heart. Serum glucose levels were measured
by glucose oxidase method (Venkatesh et al., 2004).

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For Paederia foetida stem methanol extract, mice were divided into six groups of ten mice each. The
various groups received different treatments like Group-I received vehicle (1% Tween 80 in water, 10 ml/kg
body weight) and served as control, group-II received standard drug (glibenclamide, 10 mg/kg body weight)
and the other four groups (III-VI) received the methanol extract of Paederia foetida stems at four different
doses of 50, 100, 250 and 500 mg/kg body weight. The rest of the experiment was conducted as described
previously for Madhuca indica leaves.
Statistical Analysis:
Experimental values are expressed as mean ± SEM. Independent Sample t-test was carried out for
statistical comparison. Statistical significance was considered to be indicated by a p value < 0.05 in all cases.
Results and discussion
The crude methanolic extract of Madhuca indica leaves demonstrated dose-dependent reductions in serum
glucose level following administration in glucose-loaded mice. Of the four doses of the extract tested, namely
50, 100, 250, and 500 mg extract per kg body weight, the decreases in serum glucose levels were found to
be significantly reduced at doses of 100, 250, and 500 mg extract per kg body weight. At these doses, the
extract reduced serum glucose levels, respectively, by 22.2, 25.8 and 36.3%. In comparison, the standard
antihyperglycemic drug, glibenclamide, reduced serum glucose concentration by 35.9%, when administered at
a dose of 10 mg per kg body weight. Thus the highest dose of the extract was equivalent to glibenclamide
in terms of ability to reduce serum glucose levels. The results are shown in Table 1.
The crude methanolic extract of stems of Paederia foetida also demonstrated serum glucose reducing
ability in a dose-dependent manner. At the four doses of extract tested, namely 50, 100, 250, and 500 mg
extract per kg body weight, serum glucose levels were reduced by 7.7, 25.3, 31.0, and 31.2%, respectively.
Essentially, the activity of the extract peaked at a dose of 250 mg extract administered per kg body weight
of mice. The results are shown in Table 2. Overall, the conclusion from both experiments was that the leaf
extract of Madhuca indica and stem extract of Paederia foetida had strong antihyperglycemic effects in
glucose-loaded mice and thus can be considered for further studies towards isolation of antidiabetic compounds.
Other Sapotaceae and Rubiaceae family (to which Madhuca indica and Paederia foetida, respectively
belongs) plants have been reported in the scientific literature for presence of antidiabetic activities in extracts
of whole plants or plant parts. As an instance, the fruits of Synsepalum dulcificum (Schumach. & Thonn.)
Daniell (Sapotaceae) showed improvement in insulin resistance in fructose-rich chow-fed rats and decreased
plasma glucose concentrations (Chen et al., 2006). Aqueous leaf extract of Rothmannia longiflora Salisb.
(Rubiaceae) has been shown to improve basal metabolic rate and electrolyte parameters in alloxan-induced
diabetic rats (Ikpi et al., 2009). a-Glucodidase inhibitors have been reported from the plant, Luculia pinciana
Hook. (Rubiaceae) (Kang et al., 2009). Antidiabetic activity has been reported for ethanolic leaf extract of
Heinsia crinita (Afzel.) G. Taylor (Rubiaceae) in alloxan-induced diabetic rats (Okokon et al., 2009). Extract
of leaves of Hintonia standleyana Bullock (Rubiaceae) and Hintonia latiflora (DC.) Bullock (Rubiaceae) has
been found to decrease blood glucose levels in both normal and streptozotocin-induced diabetic rats (Cristians
et al., 2009).
Table 1: Effect of methanol extract of Madhuca indica leaves on serum glucose level in hyperglycemic mice following 120 minutes of
glucose loading.
Treatment Dose (mg/kg body weight) Serum glucose level (mg/dl) % lowering of serum glucose level
Control 10 ml 87.94 ± 5.56 -
Glibenclamide 10 mg 56.39 ± 8.01 35.9*
Madhuca indica 50 mg 88.65 ± 4.58 -
Madhuca indica 100 mg 68.44 ± 6.49 22.2*
Madhuca indica 250 mg 65.25 ± 8.08 25.8*
Madhuca indica 500 mg 56.03 ± 6.85 36.3*
All administrations were made orally. Values represented as mean ± SEM, (n=6); *P < 0.05; significant compared to hyperglycemic
control animals.
Table 2: Effect of methanol extract of Paederia foetida stems on serum glucose level in hyperglycemic mice following 120 minutes of
glucose loading.
Treatment Dose (mg/kg body weight) Serum glucose level (mg/dl) % lowering of serum glucose level
Control 10 ml 86.15 ± 5.24 -
Glibenclamide 10 mg 51.79 ± 4.78 39.9*
Paederia foetida 50 mg 79.48 ± 5.07 7.7
Paederia foetida 100 mg 64.36 ± 3.51 25.3*
Paederia foetida 250 mg 59.48 ± 3.70 31.0*
Paederia foetida 500 mg 59.25 ± 5.15 31.2*
All administrations were made orally. Values represented as mean ± SEM, (n=6); *P < 0.05; significant compared to hyperglycemic
control animals.

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The antihyperglycemic activity exhibited by any extract or compound may occur through several
mechanisms. The extract may potentiate the pancreatic secretion of insulin or increase the glucose uptake
(Farjou et al., 1987; Nyunai et al., 2009). Alternative mechanisms behind demonstration of any
antihyperglycemic effect include inhibition of glucose absorption in gut (Bhowmik et al., 2009), or restoration
of damaged pancreatic islets, which caused decreased secretion of insulin in the first place (Venkatesh et al.,
2010). The exact mechanism(s) involved behind the antihyperglycemic activities shown by Madhuca indica
leaves and Paederia foetida stems are currently being investigated. However, the results strongly indicate the
importance of further studies on these two plants towards discovery of possible novel antidiabetic agents.
Notably, diabetes is a disease affecting millions of people throughout the world currently, and for which
modern medicine has no known cure.
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