An evaluation of antihyperglycemic and antinociceptive effects of methanol extract of Cassia fistula L. (Fabaceae) leaves in Swiss albino mice.
Cassia fistula L. (local name: shonalu, English name: golden shower tree) is a member of the Fabaceae family and is native to southern Asia. It is common in Bangladesh, being found both in forest areas or else planted by the roadsides as an ornamental tree because of its abundant and showy flowers. The leaves and other parts of the plant are used by the folk medicinal practitioners (Kavirajes) of Bangladesh for treatment of diabetes, pain, fever, helminthiasis, and stomach disorders. In Indian traditional medicine, the plant is also used for the treatment of diabetes.
Catechin has been isolated from methanol extract of stems of the plant, and which has been shown to have insulin mimetic effects on glucose oxidation and glucose uptake in streptozotocin-induced diabetic Wistar rats (Daisy et al., 2010). The alcohol extract of leaves of the plant also reportedly showed wound healing properties as demonstrated by better wound closure and improved tissue regeneration at the wound site of rats (Senthil Kumar et al., 2006). Besides Cassia fistula, other plants belonging to the same genus have been reported to have diverse pharmacological activities, including antidiabetic and antinociceptive activities. The aqueous extract of Cassia occidentalis has been shown to exhibit significant antihyperglycemic activity in normal and alloxan-induced diabetic rats (Verma et al. , 2010). An active principle, Cg-1, has been isolated from Cassia glauca leaves, which demonstrated both cardioprotective potential as well as antidiabetic activity in diabetic rats (Mazumder et al., 2009). Aqueous extract of bark of Cassia glauca demonstrated significant antidiabetic potential in ameliorating the diabetic condition in streptozotocin-induced diabetic rats (Salahuddin et al., 2010). Antidiabetic effect has also been observed with Cassia auriculata leaf extract in streptozotocin-induced diabetic rats (Gupta et al. , 2010). The antihyperglycemic and hypolipidemic activity of aqueous extract of Cassia auriculata leaves in experimental diabetes in rat model has been shown (Gupta et al., 2009). Leaf extract of Cassia alata was reported to be effective in reducing blood sugar levels in streptozotocin-induced hyperglycemic rats (Palanichamy et al., 1988).
Hexane and ethyl acetate extracts of Cassia alata leaves have been reported to demonstrate antiinflammatory activities; additionally, the chloroform extract of leaves was found to be antimutagenic, while the ethyl acetate extract exhibited hypoglycemic properties (Villasenor et al., 2002). The antinociceptive profile of (-)-spectaline, a piperidine alkaloid isolated from Cassia spectabilis and Cassia leptophylla has been reported (Viegas et al., 2008; Alexandre-Moreira et al., 2003). Methanol extract of root bark of Cassia singueana reportedly demonstrated antinociceptive, antipyretic and antiplasmodial activity in mice and rat models (Adzu et al., 2003). Antinociceptive and smooth muscle contracting activities of the methanolic extract of Cassia tora leaves has been described (Chidume et al., 2002). Considering the fact that a number of members of the Cassia genus have been reported in the scientific literature for their antidiabetic as well as antinociceptive activities, and that Cassia fistula is used by the folk medicinal practitioners of bangladesh for treatment of diabetes and pain, it was the objective of the present study to evaluate the antihyperglycemic and antinociceptive potential of methanol extracts of leaves of this plant in rodent model systems.
Materials and methods
Plant Material and Extraction:
The leaves of Cassia fistula L. were collected from Dhaka district, Bangladesh in July, 2009. The plant was taxonomically identified by Bangladesh National Herbarium at Dhaka (Accession Number 35,155). The leaves of Cassia fistula 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 6.7g.
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.
In the present study, Swiss albino mice (male), which weighed between 20-25g 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 one week prior to actual experiments. The study was conducted following approval by the Institutional Animal Ethical Committee of University of Development Alternative, Dhaka, Bangladesh.
Glucose tolerance property of methanol extract of Cassia fistula leaves was determined as per the procedure previously described by Joy and Kuttan (1999) with minor modifications. In brief, fasted mice were grouped into six groups of seven 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 three groups (III-V) received the methanol extract of Cassia fistula leaves at four different doses of 50, 100, 200 and 400 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 the glucose administration through puncturing heart. Serum glucose levels were measured by glucose oxidase method (Venkatesh et al., 2004).
Acetic Acid-induced Writhing Method:
Antinociceptive activity of methanol extract of Cassia fistula leaves was examined using previously described procedures of Deb et al. (2010) with minor modifications. Pain was induced in mice in the writhing test through intraperitoneal administration of 1% acetic acid at a dose of 10 ml/kg body weight. Mice were separated into six groups of six mice each. Group-I served as control and was administered vehicle (1% Tween 80 in water, 10 ml/kg body weight). The standard drug, aspirin was administered to Group-II mice at a dose of 200 mg/kg body weight. Groups-III to VI received extract, respectively at 50, 100, 200, and 400 mg extract/kg body weight orally 30 min before acetic acid injection. A period of 5 minutes was given to each animal to ensure bio-availability of acetic acid, following which period, the number of writhings was counted for 10 min.
Experimental values are expressed as mean [+ or -] SEM. Independent Sample t-test was carried out for statistical comparison. Statistical significance was considered to be indicated byap value < 0.05 in all cases.
Results and discussion
The results from the study showed that the methanol extract of leaves of Cassia fistula dosedependently and significantly lowered serum glucose levels in glucose-loaded hyperglycemic mice. At the lowest dose tested, namely that of 50 mg/kg body weight, serum glucose levels in glucose-loaded mice was lowered by 24.7%; however, the results were not significant when compared with control mice. At the higher doses tested of the extract, i.e. 100, 200 and 400 mg/kg body weight, serum glucose levels were lowered, respectively, by 26.9, 32.1 and 47.0%, which were significantly less than that of control animal values. In comparison, the standard antihyperglycemic drug, glibenclamide, when administered at a dose of 10 mg/kg body weight also lowered serum glucose levels in mice by 47.0%, which was the same value obtained with the highest dose (400 mg) of the extract tested. The results are shown in Table 1. From the results obtained, it can be concluded that the methanol extract of leaves possessed significant antihyperglycemic activity and can be considered for further studies towards isolation of phytochemical constituent(s) leading to discovery of possible antidiabetic drugs.
Several mechanisms may be relevant factors behind the observed decrease in serum glucose levels in mice administered with the methanol extract of the plant. The extract may potentiate the pancreatic secretion of insulin or increase the glucose uptake (Nyunai et al., 2009; Farjou et al., 1987).
Alternately, the extract may inhibit glucose absorption in gut (Bhowmik et al., 2009). The exact mechanism through which the extract lowered serum glucose levels is presently being studied in our laboratory.
The methanol extract of leaves of Cassia alata also demonstrated a dose-dependent reduction in acetic acid writhings in mice. The results are shown in Table 2. At doses, respectively, of 50, 100, 200 and 400 mg extract/kg body weight, percent inhibitions of writhings were 7.9, 13.8, 21.5 and 60.8. However, the results were significant only at the highest dose (400 mg) of the extract tested. In comparison, the standard antinociceptive drug, aspirin, caused a 41.2% inhibition in writhings. As such, at the highest dose tested of the extract, considerably more antinociceptive activity was obtained than obtained with aspirin.
Both central and peripheral analgesia can be detected with the acetic acid-induced writhing test (Shanmugasundaram and Venkataraman, 2005). Production of prostaglandins [mainly prostacyclines ([PGI.sub.2]) and prostaglandin- (PG-E)] has been shown to be responsible for excitation of Ad-nerve fibers, leading to the sensation of pain (Reynolds, 1982; Rang and Dale, 1993). The mechanism behind intraperitoneal administration of acetic acid and subsequent evolvement of pain leading to gastric writhings, probably then involve a mechanism where increased production of prostaglandins is involved. It is then possible that the mechanism behind the inhibition of gastric writhings by the extract is mediated through inhibition of prostaglandin synthesis. It may be seen from Table 2, that the extract when administered at a dose of 400 mg/kg body weight reduced abdominal constrictions or writhings in mice, considerably more than the standard drug, aspirin. The obtained results are suggestive that the extract may contain phytochemical components, which following administration has lead to inhibition of lipooxygenase and/or cyclooxygenase. This inhibition will lead in turn to reduction of prostaglandin E2 synthesis, thereby causing amelioration or cessation of pain caused by intraperitoneal administration of acetic acid. Notably, a similar mechanism has been proposed for antinociceptive activity of Ficus deltoidea aqueous extract in acetic acid-induced gastric pain model (Sulaiman et al., 2008). Studies are being conducted in our laboratory to identify the constituents of the methanol extract of Cassia fistula responsible for the antinociceptive effect observed in acetic acid-injected mice.
Bangladesh has a rich diversity of medicinal plants, which we have been studying for the last few years. Folk medicinal uses of plants has continued in Bangladesh from time immemorial and are still very much in existence today, as documented by a number of ethnomedicinal papers from our laboratory (Rahmatullah et al., 2009 a-e). Pharmacological activity studies, conducted in our laboratory, has also validated the traditional uses of a number of medicinal plants obtained from our various ethnomedicinal surveys (Morshed et al., 2010; Ahmed et al., 2010; Moushumi et al., 2010). The present study also validates the folk medicinal uses of Cassia fistula for treatment of diabetes and pain. The plant, following further scientific studies and clinical trials, can thus form a cost-effective solution as an antidiabetic and an antinociceptive agent to the vast majority of the people of Bangladesh, who either cannot afford modern allopathic medicines or lacks access to modern health-care facilities.
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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. Email: email@example.com Telephone: +88-01715032621 Fax: +88-02-815739
(1) Zahidul Islam Khan, (1) Badrun Nahar, (1) Md. Abu Jakaria, (1) Shahnaz Rahman, (2) Majeedul H. Chowdhury, (1) Mohammed Rahmatullah
(1) Department of Biotechnology & Genetic Engineering, University of Development Alternative, Dhanmondi, Dhaka-1205, Bangladesh
(2) Present address: New York City College of Technology The City University of New York 300 Jay Street, Brooklyn, NY11201, USA.
(1) Zahidul Islam Khan, (1) Badrun Nahar, (1) Md. Abu Jakaria, (1) Shahnaz Rahman, (2) Majeedul H. Chowdhury, (1) Mohammed Rahmatullah: An Evaluation of Antihyperglycemic and Antinociceptive Effects of Methanol Extract of Cassia Fistula L. (Fabaceae) Leaves in Swiss Albino Mice
Table 1: Effect of methanol extract of Cassia fistula leaves on serum glucose level in hyperglycemic mice. % lowering of Dose (mg/kg Serum glucose serum glucose Treatment body weight) level (mg/dl) level Control 10 ml 132.23 [+ or -] 16.01 -- Glibenclamide 10 mg 69.96 [+ or -] 6.13 47.0 * Cassia fistula 50 mg 99.63 [+ or -] 12.98 24.7 Cassia fistula 100 mg 96.70 [+ or -] 3.05 26.9 * Cassia fistula 200 mg 89.74 [+ or -] 7.59 32.1 * Cassia fistula 400 mg 69.96 [+ or -] 5.53 47.0 * All administrations were made orally. Values represented as mean [+ or -] SEM, (n=7); * P < 0.05; significant compared to hyperglycemic control animals. Table 2: Antinociceptive effect of crude methanol extract of Cassia fistula leaves in the acetic acid-induced gastric pain model mice. Dose (mg/kg Mean no. Groups body weight) of writhing Inhibition (%) Control (vehicle) 10 ml 8.5 [+ or -] 0.7 -- Aspirin 200 mg 5.0 [+ or -] 1.2 41.2 * Cassia fistula 50 mg 7.8 [+ or -] 1.9 7.9 Cassia fistula 100 mg 7.3 [+ or -] 2.7 13.8 Cassia fistula 200 mg 6.7 [+ or -] 2.7 21.5 Cassia fistula 400 mg 3.3 [+ or -] 1.4 60.8 *
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|Title Annotation:||Original Article|
|Author:||Khan, Zahidul Islam; Nahar, Badrun; Jakaria, Abu; Rahman, Shahnaz; Chowdhury, Majeedul H.; Rahmatull|
|Publication:||Advances in Natural and Applied Sciences|
|Date:||Sep 1, 2010|
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