Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Advances in Natural and Applied Sciences, 7(5) December 2013, Pages: 519-525 AENSI Journals Advances in Natural and Applied Sciences Journal home page: www.aensiweb.com/anas/index.html Antihyperglycemic activities of leaves of Brassica oleracea, Centella asiatica and Zizyphus mauritiana: Evaluation through oral glucose tolerance tests Sadia Haque, Tania Naznine, Mohammad Ali, Tania Tabassum Azad, Md. Tanvir Morshed, Nusrat Anik Afsana, Ishtiaq Ahmed, Mohammed Rahmatullah Faculty of Life Sciences, University of Development Alternative, Dhanmondi, Dhaka-1205, Bangladesh. ARTICLE INFO Article history: Received 14 November 2013 Received in revised form 24 December 2013 Accepted 28 December 2013 Available online 15 February2014 Key words: Brassica oleracea, Centella asiatica, Zizyphus mauritiana, antihyperglycemic, OGTT ABSTRACT The antihyperglycemic potentials of methanolic extract of leaves of Brassica oleracea, Centella asiatica, and Zizyphus mauritiana were evaluated by oral glucose tolerance tests in glucose loaded Swiss albino mice. At doses of 50, 100, 200 and 400 mg per kg body weight, administration of extract of Brassica oleracea leaves led to reductions in blood glucose levels in glucose-loaded mice by, respectively, 22.4, 24.2, 34.0, and 47.6%. The results were both dose-dependent, as well as statistically significant. At the afore-mentioned four doses, administration of extract of leaves of Centella asiatica led to dose-dependent and significant reductions in blood glucose levels in mice by 29.4, 32.8, 33.6, and 35.7%, respectively. At the same four doses, administration of extract of leaves of Zizyphus mauritiana lead to dose-dependent and significant reductions in blood glucose levels by 37.8, 38.9, 40.5, and 43.1%, respectively. A standard antihyperglycemic drug, glibenclamide, when administered at a dose of 10 mg per kg body weight, led to reductions in blood sugar levels in glucose-loaded mice within a range of 41.3 – 54.2%. It can be concluded that the leaves of all three plants possess significant antihyperglycemic effects and can be utilized as potential sources for new antidiabetic drugs, or used as crude remedies for lowering of blood sugar in diabetic patients. © 2013 AENSI Publisher All rights reserved. To Cite This Article: Sadia Haque, Tania Naznine, Mohammad Ali, Tania Tabassum Azad, Md. Tanvir Morshed, Nusrat Anik Afsana, Ishtiaq Ahmed, Mohammed Rahmatullah., Antihyperglycemic activities of leaves of Brassica oleracea, Centella asiatica and Zizyphus mauritiana: Evaluation through oral glucose tolerance tests. Adv. in Nat. Appl. Sci., 7(5): 519-525, 2013 INTRODUCTION Brassica oleracea L. var. gongylodes (Family: Cruciferae; English: kohlrabi; Bengali: ol kopi) is a winter crop of Bangladesh and belongs to the same genera as other more well-known species like cabbage, cauliflower or broccoli. The whole plant is consumed in the cooked form in Bangladesh, but more often the swollen portion of the plant below the ground is consumed. The leaves of Centella asiatica (L.) Urb. (Family: Apiaceae; English: Asiatic pennywort; Bengali: thankuni) are also consumed in the cooked form as soup or as a vegetable dish in Bangladesh, more so during episodes of gastrointestinal disorders like diarrhea, dysentery, bloating, and indigestion. Occasionally, the plant is also consumed to increase memory. The fruits of Zizyphus mauritiana Lam. (Family: Rhamnaceae; English: Indian plum; Bengali: boroi) are also consumed raw when ripe in Bangladesh or taken in the form of pickles or chutney. Diabetes is a disease affecting millions of people throughout the world and is manifested primarily by increases in blood sugar levels, and in later stages can lead to complicated disorders like cardiovascular problems, diabetic retinopathy and diabetic nephropathy. The disease has no known total cure in allopathic medicine, but traditional medicinal practitioners of many countries, including Bangladesh, claim to have plant remedies that can totally cure the disease. The leaves of the above three plants are often advised by the folk medicinal practitioners of Bangladesh to be consumed in the form of decoction or in the form of cooked vegetables for lowering of blood sugar in diabetic patients. According to projections made by the World Health Organization, the number of diabetic patients in the world may increase by 35% within 2025 (Boyle et al., 2001). As such, it is extremely important to find a total cure for diabetes before it reaches endemic proportions. Various members of the Brassica oleracea family have been reported to have antihyperglycemic effects. The antihyperglycemic and glycogenesis effects of cabbage (Brassica oleracea L. var. capitata) have been reported in alloxan-induced diabetic rats (Asaduzzaman et al., 2011). Juice obtained from cabbage leaves is also Corresponding Author: Dr. Mohammed Rahmatullah, Pro-Vice Chancellor and Dean, Faculty of Life Sciences University of Development Alternative House No. 78, Road No. 11A (new) Dhanmondi, Dhaka-1205 Bangladesh Tel: +88-01715032621; Fax: +88-02-815739; E-mail: [email protected] 520 Dr. Mohammed Rahmatullah et al, 2013 Advances in Natural and Applied Sciences, 7(5) December 2013, Pages: 519-525 used as traditional medicine for treatment of diabetes by inhabitants of Douala Town, Cameroon (Din et al., 2011). Aqueous extract of cabbage has also been shown to improve a number of parameters in diabetic rats (Luka and Tijjani, 2013). The antidiabetic effect of Korean traditional Baechu (Chinese cabbage) kimchi has been shown in type 2 diabetic rats (Islam and Choi, 2009). Amelioration of diabetic nephropathy by extract of red cabbage has been reported (Kataya and Hamza, 2007). The antidiabetic effect of ethanol extract of Centella asiatica leaves in alloxan diabetic rats has been reported (Rahman et al., 2012). The plant juice has also been shown to exert antidiabetic activity in Type 1 model of alloxan induced diabetic rats (Rahman et al., 2011). In glucose tolerance tests, ethanolic and methanolic extract of leaves normalized glucose levels in alloxan induced diabetic rats (Chauhan et al., 2010). Anti-diabetic activity of ethanol extract of whole plant has been shown in streptozotocin-induced diabetic rats (Gayathri et al., 2011). The hypoglycemic activity of Ziziphus mauritiana aqueous-ethanol seed extract in alloxan-induced diabetic mice has been demonstrated (Bhatia and Mishra, 2010). Various solvent extracts of fruits of the plant also reportedly showed antihyperglycemic activity in glucose overloaded hyperglycemic rats (Jarald et al., 2009). Studies conducted by our research group have centered on ethnomedicinal surveys (Rahmatullah et al., 2009a-c; Rahmatullah et al., 2010a-g; Rahmatullah et al., 2011a,b; Rahmatullah et al., 2012a-d), followed by screening of the plants obtained for antihyperglycemic, antinociceptive and cytotoxic activities (Anwar et al., 2010; Jahan et al., 2010; Khan et al., 2010; Mannan et al., 2010; Rahman et al., 2010; Rahmatullah et al., 2010h; Shoha et al., 2010; Ali et al., 2011; Barman et al., 2011; Hossan et al., 2011; Jahan et al., 2011; Rahman et al., 2011; Sutradhar et al., 2011; Ahmed et al., 2012; Arefin et al., 2012; Haque et al., 2012; Sathi et al., 2012). As part of the screening process to locate plants with antihyperglycemic properties, this study was conducted to evaluate the antihyperglycemic potential of methanol extract of leaves of Brassica oleracea, Centella asiatica, and Zizyphus mauritiana in oral glucose tolerance tests (OGTT) in glucose-overloaded mice. MATERIALS AND METHODS Leaves of Brassica oleracea were collected from Dhaka district, Bangladesh during January, 2013. The plant was taxonomically identified at the Bangladesh National Herbarium at Dhaka (Accession Number 38,099). The sliced and air-dried leaves of Brassica oleracea were grounded into a fine powder and 100g of the powder was extracted with methanol (500 ml) for 48 hours. The extract was evaporated to dryness. The final weight of the extract was 3g. Leaves of Centella asiatica were collected from Dhaka district, Bangladesh during May, 2013. The plant was taxonomically identified at the Bangladesh National Herbarium at Dhaka (Accession Number 38,359). The sliced and air-dried leaves of Centella asiatica were grounded into a fine powder and 57g of the powder was extracted with methanol (300 ml) for 48 hours. The extract was evaporated to dryness. The final weight of the extract was 4.35g. Leaves of Zizyphus mauritiana were collected from Dhaka district, Bangladesh during June, 2013. The plant was taxonomically identified at the Bangladesh National Herbarium at Dhaka (Accession Number 38,560). The sliced and air-dried leaves of Ziziphus mauritiana were grounded into a fine powder and 100g of the powder was extracted with methanol (500 ml) for 48 hours. The extract was evaporated to dryness. The final weight of the extract was 5g. Chemicals: Glacial acetic acid was obtained from Sigma Chemicals, USA; aspirin, glibenclamide and glucose were obtained from Square Pharmaceuticals Ltd., Bangladesh. Animals: In the present study, Swiss albino mice (male), which weighed between 15-22 g 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. Antihyperglycemic activity: Glucose tolerance property of the various extracts 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 six mice each for Brassica oleracea, six groups of five mice each for Centella asiatica, and six groups of five mice each for Zizyphus mauritiana. The various groups received different treatments like Group 1 received vehicle (1% Tween 80 in water, 10 ml/kg body weight) and served as control, group 2 received standard drug (glibenclamide, 10 mg/kg body weight). Groups 3-6 received methanol extract of leaves at doses of 50, 100, 200 and 400 mg per 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. 521 Dr. Mohammed Rahmatullah et al, 2013 Advances in Natural and Applied Sciences, 7(5) December 2013, Pages: 519-525 Following a period of one hour, all mice were orally administered 2 g glucose/kg of body weight. Blood samples were collected 120 minutes after the glucose administration through puncturing heart. Blood glucose levels were measured by glucose oxidase method (Venkatesh et al., 2004). 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 At doses of 50, 100, 200 and 400 mg per kg body weight, administration of extract of Brassica oleracea leaves led to reductions in blood glucose levels in glucose-loaded mice by, respectively, 22.4, 24.2, 34.0, and 47.6%. The results were both dose-dependent, as well as statistically significant. In comparison, a standard antihyperglycemic effect, glibenclamide, when administered at a dose of 10 mg per kg body weight, reduced blood glucose level by 41.3%. Thus at the highest dose of the Brassica oleracea extract tested (400 mg), the extract demonstrated better antihyperglycemic potential than the standard drug, glibenclamide. The results are shown in Table 1. At the afore-mentioned four doses, i.e. 50, 100, 200 and 400 mg per kg body weight, administration of extract of leaves of Centella asiatica led to dose-dependent and significant reductions in blood glucose levels in mice by 29.4, 32.8, 33.6, and 35.7%, respectively. Glibenclamide, when administered at a dose of 10 mg per kg body weight, reduced blood glucose level in this group of mice by 48.9%. The results are shown in Table 2. At the same four doses, administration of extract of leaves of Zizyphus mauritiana lead to dose-dependent and significant reductions in blood glucose levels by 37.8, 38.9, 40.5, and 43.1%, respectively. With this group of mice, glibenclamide reduced the blood glucose level by 54.2%. The results are shown in Table 3. Notably, the effect of glibenclamide varied somewhat from group experiment to group experiment, but nevertheless fell within the range of 41.3 – 54.2%. This is not unusual, given that the weight of mice varied between the three experiments conducted with the three different extracts. But overall it can be concluded that all extracts demonstrated significant antihyperglycemic potential. Several mechanisms can be attributed to the observed lowering of blood glucose by the extracts in the present study. Glucose absorption in gut may be inhibited by a compound or compounds, as observed with Mangifera indica L. (Anacardiaceae) stem-barks (Bhowmik et al., 2009). Another possible mechanism can possibly be increase of peripheral glucose consumption induced by the extract, as has been seen with ethanolic extract of Sapindus trifoliatus L. (Sapindaceae) (Sahoo et al., 2010). Alternately, antihyperglycemic compound or compounds present in the extract may lower blood sugar either through potentiating the pancreatic secretion of insulin or increasing the glucose uptake, as has been observed in studies with Artemisia extract and extract of Ageratum conyzoides L. (Asteraceae), respectively (Farjou et al., 1987; Nyunai et al., 2009). The exact mechanism for lowering of blood glucose by the three extracts has not been determined in the present study and is currently ongoing in our laboratory. Brassica oleracea var. gongylodes has been reported to contain quercetin (Duke, 1992). The anti-diabetic effect of quercetin has been confirmed in rats (Abdelmoaty et al., 2010). Centella asiatica is known to contain a number of bio-active compounds, including kaempferol and quercetin derivatives, brahmoside, brahminoside, beta-sitosterol, and kaempferol (Duke, 1992). The hypoglycemic and antioxidant potential of kaempferol-3,7(O)-alpha-dirhamnoside has been shown (de Sousa et al., 2004). Beta-sitosterol also has been shown to have anti-diabetic efficacy through lowering of blood glucose levels, which has been attributed to its ability to increase peripheral glucose consumption (Karan et al., 2012). Brahmoside and brahminoside has been shown to exert hypoglycemic effects in rats (Ramaswamy, 1970), which may also explain the observed antihyperglycemic action of Centella asiatica leaf extract. Zizyphus mauritiana reportedly contains betulinic acid (Duke, 1992). Betulinic acid has been shown to be a component of Morus alba root bark extract, which exerted a hypoglycemic effect in streptozotocin-induced diabetic rats (Singab et al., 2005). Betulinic acid reportedly is also an alpha-glucodidase inhibitor, which may explain its hypoglycemic action (Benalla et al., 2010). The beneficial effect of pentacyclic triterpenoids like betulinic acid in diabetes and diabetes-induced complications has been reviewed (Alqahtani et al., 2013). Taken together, the results obtained in the present study suggest that consumption of Brassica oleracea and Centella asiatica may prove beneficial to diabetic patients, when consumed on a regular basis. Studies are ongoing in our laboratory for identification of the responsible phytochemicals in leaves of all three plants for the observed antihyperglycemic effects, and the mechanisms through which the phytochemicals are exerting their antihyperglycemic actions. 522 Dr. Mohammed Rahmatullah et al, 2013 Advances in Natural and Applied Sciences, 7(5) December 2013, Pages: 519-525 Table 1: Effect of methanol extract of Brassica oleracea leaves on blood glucose level in hyperglycemic mice following 120 minutes of glucose loading. Treatment Dose (mg/kg body Blood glucose level (mmol/l) % lowering of blood glucose level weight) Control (Group 1) 10 ml 5.50 ± 0.31 Glibenclamide (Group 2) 10 mg 3.23 ± 0.14 41.3* Brassica oleracea (Group 3) 50 mg 4.27 ± 0.43 22.4* Brassica oleracea (Group 4) 100 mg 4.12 ± 0.32 24.2* Brassica oleracea (Group 5) 200 mg 3.63 ± 0.35 34.0* Brassica oleracea (Group 6) 400 mg 2.88 ± 0.28 47.6* 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 Centella asiatica leaves on blood glucose level in hyperglycemic mice following 120 minutes of glucose loading. Treatment Dose (mg/kg body Blood glucose level (mmol/l) % lowering of blood weight) glucose level Control (Group 1) 10 ml 4.70 ± 0.38 Glibenclamide (Group 2) 10 mg 2.40 ± 0.21 48.9* Centella asiatica (Group 3) 50 mg 3.32 ± 0.33 29.4* Centella asiatica (Group 4) 100 mg 3.16 ± 0.26 32.8* Centella asiatica (Group 5) 200 mg 3.12 ± 0.20 33.6* Centella asiatica (Group 6) 400 mg 3.02 ± 0.33 35.7* All administrations were made orally. Values represented as mean ± SEM, (n=5); *P < 0.05; significant compared to hyperglycemic control animals. Table 3: Effect of methanol extract of Zizyphus mauritiana leaves on blood glucose level in hyperglycemic mice following 120 minutes of glucose loading. Treatment Dose (mg/kg body Blood glucose level (mmol/l) % lowering of blood weight) glucose level Control (Group 1) 10 ml 5.24 ± 0.37 Glibenclamide (Group 2) 10 mg 2.40 ± 0.21 54.2* Zizyphus mauritiana (Group 3) 50 mg 3.26 ± 0.24 37.8* Zizyphus mauritiana (Group 4) 100 mg 3.20 ± 0.29 38.9* Zizyphus mauritiana (Group 5) 200 mg 3.12 ± 0.30 40.5* Zizyphus mauritiana (Group 6) 400 mg 2.98 ± 0.18 43.1* All administrations were made orally. Values represented as mean ± SEM, (n=5); *P < 0.05; significant compared to hyperglycemic control animals. REFERENCES Abdelmoaty, M.A., M.A. Ibrahim, N.S. Ahmed and M.A. Abdelaziz, 2010. Confirmatory studies on the antioxidant and antidiabetic effect of quercetin in rats. Indian Journal of Clinical Biochemistry, 25: 188-192. Ahmed, T., K.M.S.U. Imam, S. Rahman, S.M. Mou, M.S. Choudhury, M.J. Mahal, S. Jahan, M.S. Hossain and M. Rahmatullah, 2012. Antihyperglycemic and antinociceptive activity of Fabaceae family plants – an evaluation of Mimosa pigra L. stems. Advances in Natural and Applied Sciences, 6: 1490-1495. Ali, M., K. Nahar, M. Sintaha, H.N. Khaleque, F.I. Jahan, K.R. Biswas, A. Swarna, M.N. Monalisa, R. Jahan and M. Rahmatullah, 2011. An evaluation of antihyperglycemic and antinociceptive effects of methanol extract of Heritiera fomes Buch.-Ham. (Sterculiaceae) barks in Swiss albino mice. Advances in Natural and Applied Sciences, 5: 116-121. Alqahtani, A., K. Hamid, A. Kam, K.H. Wong, Z. Abdelhak, V. Razmovski-Naumovski, K. Chan, K.M. Li, P.W. Groundwater and G.Q. Li, 2013. The pentacyclic triterpenoids in herbal medicines and their pharmacological activities in diabetes and diabetic complications. Current Medicinal Chemistry, 20: 908-931. Anwar, M.M., M.A. Kalpana, B. Bhadra, S. Rahman, S. Sarker, M.H. Chowdhury and M. Rahmatullah, 2010. Antihyperglycemic activity and brine shrimp lethality studies on methanol extract of Cajanus cajan (L.) Millsp. leaves and roots. Advances in Natural and Applied Sciences, 4: 311-316. Arefin, S.A., S. Rahman, S. Rahman, M. Akter, M. Munmun, M.A. Kalpana, S. Jahan, M.S.A. Bhuiyan, and M. Rahmatullah, 2012. Scientific validation of folk medicinal uses in Bangladesh of Piper betle leaves to alleviate pain and lower blood sugar. Advances in Natural and Applied Sciences, 6: 1496-1502. Asadujjaman, M., M.S. Hossain, M.R.I. Khan, A.S.M. Anisuzzaman, M. Ahmed and A. Islam, 2011. Antihyperglycemic and glycogenesis effects of different fractions of Brassica oleracea in alloxan induced diabetic rats.International Journal of Pharmaceutical Sciences and Research, 2: 1436-1442. Barman, M.R., Md. Saleh Uddin, S. Akhter, Md.N. Ahmed, Z. Haque, S. Rahman, F. Mostafa, M. Zaman, F.A. Noor and M. Rahmatullah, 2011. Antinociceptive activity of methanol extract of Areca catechu L. (Arecaceae) stems and leaves in mice. Advances in Natural and Applied Sciences, 5: 223-226. Benalla, W., S. Bellahcen and M. Bnouham, 2010. Antidiabetic medicinal plants as a source of alpha glucosidase inhibitors. Current Diabetes Reviews, 6: 247-254. 523 Dr. Mohammed Rahmatullah et al, 2013 Advances in Natural and Applied Sciences, 7(5) December 2013, Pages: 519-525 Bhatia, A., and T. Mishra, 2010. Hypoglycemic activity of Ziziphus mauritiana aqueous ethanol seed extract in alloxan-induced diabetic mice. Pharmaceutical Biology, 48: 604-610. Bhowmik, A., L.A. Khan, M. Akhter and B. Rokeya, 2009. Studies on the antidiabetic effects of Mangifera indica stem-barks and leaves on nondiabetic, type 1 and type 2 diabetic model rats. Bangladesh Journal of Pharmacology, 4: 110-114. Boyle, J.P., A.A. Honeycutt, K.M. Narayan, T.J. Hoerger, L.S. Geiss, H. Chen and T.J. Thompson, 2001. Projection of diabetes burden through 2050: Impact of changing demography and disease prevalence in the U.S. Diabetes Care, 24: 1936-1940. Chauhan, P.K., I.P. Pandey and V.K. Dhatwalia, 2010. Evaluation of the anti-diabetic effect of ethanolic and methanolic extracts of Centella asiatica leaves extract on alloxan induced diabetic rats. Advances in Biological Research, 4: 27-30. de Sousa, E., L. Zanatta, I. Seifriz, T.B. Creczynski-Pasa, M.G. Pizzolatti, B. Szpoganicz, and F.R.M.B. Silva, 2004. Hypoglycemic effect and antioxidant potential of kaempferol-3,7-O-(alpha)-dirhamnoside from Bauhinia forficata leaves. Journal of Natural Products, 67: 829-832. Din, N., S.D. Dibong, E.M. Mpondo, R.J. Priso, N.F. Kwin and A. Ngoye, 2011. Inventory and identification of plants used in the treatment of diabetes in Douala Town (Cameroon). European Journal of Medicinal Plants, 1: 60-73. Duke, J.A., 1992. Handbook of phytochemical constituents of GRAS herbs and other economic plants. Boca Raton, FL. CRC Press. Farjou, I.B., M. Al-Ani and S.Y. Guirgea, 1987. Lowering of blood glucose of diabetic rats by Artemisia extract. Journal of the Faculty of Medicine, 92: 137-147. Gayathri, V., P. Lekshmi and R.N. Padmanabhan, 2011. Anti-diabetes activity of ethanol extract of Centella asiatica (L.) Urban (whole plant) in streptozotocin-induced diabetic rats, isolation of an active fraction and toxicity evaluation of the extract. International Journal of Medicinal and Aromatic Plants, 1: 278-286. Haque, M.M., M.S. Choudhury, M.S. Hossain, M.A. Haque, K. Debnath, S. Hossain, S.M. Mou, I. Malek, and M. Rahmatullah, 2012. Evaluation of antihyperglycemic and antinociceptive properties of leaves of Calotropis gigantea R.Br. (Asclepiadaceae) – a medicinal plant of Bangladesh. Advances in Natural and Applied Sciences, 6: 1508-1514. Hossan, A.N.M.F., F. Zaman, M.R. Barman, S. Khatoon, M. Zaman, F. Khatun, T. Mosaiab, F. Mostafa, M. Sintaha, F. Jamal and M. Rahmatullah, 2011. Antinociceptive activity of Xanthium indicum J. Koenig ex Roxb. (Asteraceae) leaves and Leucas aspera (Willd.) Link (Lamiaceae) whole plants. Advances in Natural and Applied Sciences, 5: 214-217. Islam, Md. S., and H. Choi, 2009. Antidiabetic effect of Korean traditional Baechu kimchi in a Type 2 diabetes model of rats. Journal of Medicinal Food, 12: 292-297. Jahan, F.I., M.S. Hossain, A.A. Mamun, M.T. Hossain, S. Seraj, A.R. Chowdhury, Z. Khatun, N.Z. Andhi, M.H. Chowdhury and M. Rahmatullah, 2010. An evaluation of antinociceptive effect of methanol extracts of Desmodium gangeticum (L.) DC. stems and Benincasa hispida (Thunb.) Cogn. leaves on acetic acid-induced gastric pain in mice. Advances in Natural and Applied Sciences, 4: 365-369. Jahan, T., S. Shahreen, J. Banik, F. Islam, A.A. Mamun, R. Das, S. Rahman, S. Seraj, R. Jahan and M. Rahmatullah, 2011. Antinociceptive activity studies with methanol extracts of Ficus hispida L.f. leaves and fruits in Swiss albino mice. Advances in Natural and Applied Sciences, 5: 131-135. Jahan, F.I., M.S. Hossain, A.A. Mamun, M.T. Hossain, S. Seraj, A.R. Chowdhury, Z. Khatun, N.Z. Andhi, M.H. Chowdhury and M. Rahmatullah, 2010. An evaluation of antinociceptive effect of methanol extracts of Desmodium gangeticum (L.) DC. stems and Benincasa hispida (Thunb.) Cogn. leaves on acetic acid-induced gastric pain in mice. Advances in Natural and Applied Sciences, 4: 365-369. Jahan, T., S. Shahreen, J. Banik, F. Islam, A.A. Mamun, R. das, S. Rahman, S. Seraj, R. Jahan and Rahmatullah, M., 2011. Antinociceptive activity studies with methanol extracts of Ficus hispida L.f. leaves and fruits in Swiss albino mice. Advances in Natural and Applied Sciences, 5: 131-135. Jarald, E.E., S.B. Joshi and D.C. Jain, 2009. Antidiabetic activity of extracts and fraction of Zizyphus mauritiana. Pharmaceutical Biology, 47: 328-334. Joy, K.L., and R.J. Kuttan, 1999. Anti-diabetic activity of Picrorrhiza kurroa extract. Journal of Ethnopharmacology, 67: 143-148. Karan, S.K., S.K. Mishra, D. Pal and A. Mondal, 2012. Isolation of beta-sitosterol and evaluation of antidiabetic activity of Aristolochia indica in alloxan-induced diabetic mice with a reference to in-vitro antioxidant activity. Journal of Medicinal Plants Research, 6: 1219-1223. Kataya, H.A.H., and A.E.A. Hamza, 2007. Red cabbage (Brassica oleracea) ameliorates diabetic nephropathy in rats. Evidence-Based Complementary and Alternative Medicine, 5: 281-287. Luka, C.D., and H. Tijjani, 2013. Comparative studies of the aqueous extracts of Ocimum gratissimum, Aloe vera, Brassica oleracea and Ipomoea batatas on some biochemical parameters in diabetic rats. IOSR Journal of Pharmacy and Biological Sciences, 6: 23-29. 524 Dr. Mohammed Rahmatullah et al, 2013 Advances in Natural and Applied Sciences, 7(5) December 2013, Pages: 519-525 Nyunai, N., N. Njikam, E.H. Addennebi, J.T. Mbaford and D. Lamnaouer, 2009. Hypoglycaemic and antihyperglycaemic activity of Ageratum conyzoides L. in rats. African Journal of Traditional, Complementary and Alternative Medicines, 6: 123-130. Rahman, M., A. Siddika, B. Bhadra, S. Rahman, B. Agarwala, M.H. Chowdhury and M. Rahmatullah, 2010. Antihyperglycemic activity studies on methanol extract of Petrea volubilis L. (Verbenaceae) leaves and Excoecaria agallocha L. (Euphorbiaceae) stems. Advances in Natural and Applied Sciences, 4: 361-364. Rahman, M.M., M.N. Hasan, A.K. Das, M.T. Hossain, R. Jahan, M.A. Khatun and M. Rahmatullah, 2011. Effect of Delonix regia leaf extract on glucose tolerance in glucose-induced hyperglycemic mice. African Journal of Traditional, Complementary and Alternative Medicines, 8: 34-36. Rahman, S., M.A.H.M. Jamal, A. Parvin, Md. Mahfuz-Al-Mamun and M.R. Islam, 2011. Antidiabetic activity of Centella asiatica (L.) Urbana in alloxan induced Type 1 diabetic model rats. Journal of Bio-Sciences, 19: 23-27. Rahman, Md.M., M.S.B. Sayeed, Md.A. Haque, Md.M. Hassan and S.M.A. Islam, 2012. Phytochemical screening, antioxidant, anti-Alzheimer and anti-diabetic activities of Centella asiatica. Journal of Natural Products and Plant Resources, 2: 504-511. Rahmatullah, M., D. Ferdausi, M.A.H. Mollik, M.N.K. Azam, M.T. Rahman and R. Jahan, 2009a. Ethnomedicinal Survey of Bheramara Area in Kushtia District, Bangladesh. American Eurasian Journal of Sustainable Agriculture, 3: 534-541. Rahmatullah, M., A. Noman, M.S. Hossan, M.H. Rashid, T. Rahman, M.H. Chowdhury and R. Jahan, 2009b. A survey of medicinal plants in two areas of Dinajpur district, Bangladesh including plants which can be used as functional foods. American Eurasian Journal of Sustainable Agriculture, 3: 862-876. Rahmatullah, M., A.K. Das, M.A.H. Mollik, R. Jahan, M. Khan, T. Rahman and M.H. Chowdhury, 2009c. An Ethnomedicinal Survey of Dhamrai Sub-district in Dhaka District, Bangladesh. American Eurasian Journal of Sustainable Agriculture, 3: 881-888. Rahmatullah, M., D. Ferdausi, M.A.H. Mollik, R. Jahan, M.H. Chowdhury and W.M. Haque, 2010a. A Survey of Medicinal Plants used by Kavirajes of Chalna area, Khulna District, Bangladesh. African Journal of Traditional, Complementary and Alternative Medicines, 7: 91-97. Rahmatullah, M., M.A. Khatun, N. Morshed, P.K. Neogi, S.U.A. Khan, M.S. Hossan, M.J. Mahal and R. Jahan, 2010b. A randomized survey of medicinal plants used by folk medicinal healers of Sylhet Division, Bangladesh. Advances in Natural and Applied Sciences, 4: 52-62. Rahmatullah, M., A.A.B.T. Kabir, M.M. Rahman, M.S. Hossan, Z. Khatun, M.A. Khatun and R. Jahan, 2010c. Ethnomedicinal practices among a minority group of Christians residing in Mirzapur village of Dinajpur District, Bangladesh. Advances in Natural and Applied Sciences, 4: 45-51. Rahmatullah, M., M.A. Momen, M.M. Rahman, D. Nasrin, M.S. Hossain, Z. Khatun, F.I. Jahan, M.A. Khatun and R. Jahan, 2010d. A randomized survey of medicinal plants used by folk medicinal practitioners in Daudkandi sub-district of Comilla district, Bangladesh. Advances in Natural and Applied Sciences, 4: 99-104. Rahmatullah, M., M.A.H. Mollik, M.N. Ahmed, M.Z.A. Bhuiyan, M.M. Hossain, M.N.K. Azam, S. Seraj, M.H. Chowdhury, F. Jamal, S. Ahsan and R. Jahan, 2010e. A survey of medicinal plants used by folk medicinal practitioners in two villages of Tangail district, Bangladesh. American Eurasian Journal of Sustainable Agriculture, 4: 357-362. Rahmatullah, M., M.A.H. Mollik, M.K. Islam, M.R. Islam, F.I. Jahan, Z. Khatun, S. Seraj, M.H. Chowdhury, F. Islam, Z.U.M. Miajee and R. Jahan, 2010f. A survey of medicinal and functional food plants used by the folk medicinal practitioners of three villages in Sreepur Upazilla, Magura district, Bangladesh. American Eurasian Journal of Sustainable Agriculture, 4: 363-373. Rahmatullah, M., R. Jahan, M.A. Khatun, F.I. Jahan, A.K. Azad, A.B.M. Bashar, Z.U.M. Miajee, S. Ahsan, N. Nahar, I. Ahmad and M.H. Chowdhury, 2010g. A pharmacological evaluation of medicinal plants used by folk medicinal practitioners of Station Purbo Para Village of Jamalpur Sadar Upazila in Jamalpur district, Bangladesh. American Eurasian Journal of Sustainable Agriculture, 4: 170-195. Rahmatullah, M., Sk.Md.I. Sadeak, S.C. Bachar, Md.T. Hossain, Abdullah-al-Mamun, Montaha, N. Jahan, M.H. Chowdhury, R. Jahan, D. Nasrin, M. Rahman and S. Rahman, 2010h. Brine shrimp toxicity study of different Bangladeshi medicinal plants. Advances in Natural and Applied Sciences, 4: 163-173. Rahmatullah, M., T. Ishika, M. Rahman, A. Swarna, T. Khan, M.N. Monalisa, S. Seraj, S.M. Mou, M.J. Mahal and K.R. Biswas, 2011a. Plants prescribed for both preventive and therapeutic purposes by the traditional healers of the Bede community residing by the Turag River, Dhaka district. American Eurasian Journal of Sustainable Agriculture, 5: 325-331. Rahmatullah, M., M.N.K. Azam, M.M. Rahman, S. Seraj, M.J. Mahal, S.M. Mou, D. Nasrin, Z. Khatun, F. Islam and M.H. Chowdhury, 2011b. A survey of medicinal plants used by Garo and non-Garo traditional medicinal practitioners in two villages of Tangail district, Bangladesh. American Eurasian Journal of Sustainable Agriculture, 5: 350-357. 525 Dr. Mohammed Rahmatullah et al, 2013 Advances in Natural and Applied Sciences, 7(5) December 2013, Pages: 519-525 Rahmatullah, M., and K.R. Biswas, 2012a. Traditional medicinal practices of a Sardar healer of the Sardar (Dhangor) community of Bangladesh. Journal of Alternative and Complementary Medicine, 18: 10-19. Rahmatullah, M., A. Hasan, W. Parvin, M. Moniruzzaman, A. Khatun, Z. Khatun, F.I. Jahan and R. Jahan, 2012b. Medicinal plants and formulations used by the Soren clan of the Santal tribe in Rajshahi district, Bangladesh for treatment of various ailments. African Journal of Traditional, Complementary and Alternative Medicines, 9: 342-349. Rahmatullah, M., Z. Khatun, A. Hasan, W. Parvin, M. Moniruzzaman, A. Khatun, M.J. Mahal, M.S.A. Bhuiyan, S.M. Mou and R. Jahan, 2012c. Survey and scientific evaluation of medicinal plants used by the Pahan and Teli tribal communities of Natore district, Bangladesh. African Journal of Traditional, Complementary and Alternative Medicines, 9: 366-373. Rahmatullah, M., M.N.K. Azam, Z. Khatun, S. Seraj, F. Islam, M.A. Rahman, S. Jahan, M.S. Aziz and Jahan, R., 2012d. Medicinal plants used for treatment of diabetes by the Marakh sect of the Garo tribe living in Mymensingh district, Bangladesh. African Journal of Traditional, Complementary and Alternative Medicines, 9: 380-385. Ramaswamy, A.S., 1970. Pharmacological studies on C. asiatica. Journal of Research in Indian Medicine, 4: 160. Sahoo, P.K., K.M. Padhy, D. Pradhan, G. Tripathy, R. Bhoi, S. Pattanayak and S.K. Sahoo, 2010. Antidiabetic and antioxidant activity of ethanolic extract of Sapindus trifoliatus Linn. International Journal of Pharma and Bio Sciences, 1: 1-8. Sathi, S.I., S. Rahman, M.A. Shoyeb, K. Debnath, M.A. Haque, Z. Khatun, M.S. Hossain, M.M.R. Shelley and M. Rahmatullah, 2012. A preliminary study of the antihyperglycemic and antinociceptive potential of Tagetes patula L. (Asteraceae) stems. Advances in Natural and Applied Sciences, 6: 1515-1520. Shoha, J., H. Jahan, A.A. Mamun, M.T. Hossain, S. Ahmed, M.M. Hossain, S. Rahman, R. Jahan and M. Rahmatullah, 2010. Antihyperglycemic and antinociceptive effects of Curcuma zedoaria (Christm.) Roscoe leaf extract in Swiss albino mice. Advances in Natural and Applied Sciences, 5: 6-8. Singab, A.N., H.A. El-Beshbishy, M. Yonekawa, T. Nomura and T. Fukai, 2005. Hypoglycemic effect of Egyptian Morus alba root bark extract: effect on diabetes and lipid peroxidation of streptozotocin-induced diabetic rats. Journal of Ethnopharmacology, 100: 333-338. Sutradhar, B.K., M.J. Islam, M.A. Shoyeb, H.N. Khaleque, M. Sintaha, F.A. Noor, W. Newaz and M. Rahmatullah, 2011. An evaluation of antihyperglycemic and antinociceptive effects of crude methanol extract of Coccinia grandis (L.) J. Voigt. (Cucurbitaceae) leaves in Swiss albino mice. Advances in Natural and Applied Sciences, 5: 1-5. Venkatesh, S., G.D. Reddy, Y.S.R. Reddy, D. Sathyavathy and B.M. Reddy, 2004. Effect of Helicteres isora root extracts on glucose tolerance in glucose-induced hyperglycemic rats. Fitoterapia, 75: 364-367.