Sukardiman et al. SJIF Impact Factor 7.523 Volume 6, Issue 2, 1334-1339. Research Article ISSN 2277 7105 ANTIDIABETIC ACTIVITY OF DRY EXTRACTS OF SWIETENIA MAHAGONI SEEDS IN ALLOXAN-INDUCED DIABETIC BALB/C Sukardiman*, Novia Pegin, Herra Studiawan and Lusiana Arifianti Department of Pharmacognosy and Phytochemistry Faculty Pharmacy Airlangga University, Surabaya, Indonesia. Article Received on 20 Dec. 2016, Revised on 10 Jan. 2017, Accepted on 30 Jan. 2017 DOI: 10.20959/wjpr20172-7849 *Corresponding Author Dr. Sukardiman Department of Pharmacognosy and Phytochemistry Faculty Pharmacy Airlangga University, Surabaya, Indonesia. ABSTRACT Objective: The present study was carried out to evaluate the antidiabetic activity of dry extracts of Swietenia mahagoni seeds Methods: Antidiabetic was induced experimentally by a single intraperitoneal administration of 3,8 mg/20 g body weight of Alloxanmonohydrat in each male mice. Three days after alloxan administration, blood glucose level was measured using a glucometer. Mice with blood glucose levels above 200 mg/dl were considered diabetic and were used in this study. Then the mice were randomly divided into five experimental groups of six mice each. First group as positive control (Glibenclamide 0,06 mg/20 g body weight). Second group as negative control (CMC-Na 0.5%). Samples were given by oral route at three dosage levels (10mg / 20g, 20mg / 20g and 40mg / 20g body weight). After treatment finished, blood sampling was done by sterilizing the tail with alcohol and then nipping the tail at the start of the experiment and this was repeated after 2nd, 4th, 6th and 24th h. Results: Oral administration of dry extracts of Swietenia mahagoni seeds at doses of 10, 20 and 40 mg/20g body weight demonstrated antidiabetic activity in a dose independent manner comparable to that demonstrated by glibenclamide at 3 mg/kg body weight from two second to the twenty-fourth hour. Conclusion: From the studies it is concluded that, dry extract of Swietenia mahagoni seeds is suitable to prepare formulation for treatment of diabetic. KEYWORDS: Diabetes mellitus, Antidiabetic activity, Swietenia mahagoni seeds dry extract, alloxan. www.wjpr.net Vol 6, Issue 2, 2017. 1334
INTRODUCTION Diabetes mellitus is a metabolic disorder characterized by elevated levels of glucose in the blood (hyperglycemia). It is associated with abnormalities of the state of metabolism of carbohydrates, fats and proteins occur due to abnormalities in insulin secretion, insulin action (sensitivity), or both, of genetic factors and environmental factors and lead to chronic complications, including microvascular, macrovascular complications and chronic neuropathy. [1] The total number of people with diabetes is projected to rise from 366 million in 2011 to 552 million in 2030. Tradisonal medicine is fostered particularly in countries where scientific medicine is not accessible to large population for economic reasons. Complementary and alternative medicine does not compete with the successful main stream of scientific medicine. Users of complementary and alternative medicine commonly have chronic conditions and have tried conventional medicine but found that it has not offered a satisfactory solution, or has caused adverse effect. [2-3] Swietenia mahagoni Jacq is one of the most significant plants of the family Meliaceae. In a previous study the Swietenia mahogany seeds have antidiabetic activity. Pytochemical studies Swietenia mahogany seeds that in contains saponins, flavonoids and alkaloids, terpenoids, anthraquinone, cardiac glycosides, and volatile oils. [4] The seed of Swietenia mahagoni is a natural agonist of peroxisome-proliferator activated receptor (PPAR γ). The functions of these PPAR receptors after activation by drugs include an increase in lipid and cholesterol metabolism, adipocyte differentiation and improvement in insulin sensitivity. It has been demonstrated that PPAR γ is the receptor of the thiazolidinedione (TZD) class ligands. Among the TZD type antidiabetic drugs, Rosiglitazone and Troglitazone are potent adiopocyte-differentiating agents, which activate ap-2 gene expression in a PPARγdependent manner. [5] The formulation of dry extract seeds mahogany (Swietenia mahagoni Jacq) with the addition of cab-osil and Avicel. Therefore, this study was carried out to investigate the activity of dry extract seeds mahogany standardized against a random blood sugar level of mice (Mus musculus) were induced alloxan. MATERIAL AND METHODS Animals: Male mice of age 8 weeks and body weights ranging from 20-30 g were obtained from Animal Laboratory, Department of Pharmacognosy and Phytocemistry, Airlangga University. www.wjpr.net Vol 6, Issue 2, 2017. 1335
Chemicals: Alloxan monohydrate (Sigma Aldrich Chemical Co., US), Glibenclamide (Kimia Farma ), CMC-Sodium, Avicel, Cab-osil, Ethanol 96%, Glukometer (Easy Touch GCU Meter) and Blood Gluco-strip (Easy Touch GCU Meter). Plant material: The seeds of Swietenia mahagoni Jacq. were collected and identified by Purwodadi Botanical Garden, Pasuruan, Indonesia. Preparation of the plant dry extract The Swietenia mahagoni were seeds whie still simplicia.the dried seeds separately ground using an electric mill. The powered plant materials were kept at room temperture away from direct sunlight in closed, dry plastic bags. Thousand kilograms of each powdered plant material was extracted in 10 L etanol 96% for three times 24 h. The filtrat separated from solvent using a Buchner fannel. The filtrat was than evaporated using a rotary evaporator. The mahagony extract is added avicel: cab-osil with the ratio of 70:30 with the of comparisons dried 4:1. Induction of diabetes: The mice were injected with alloxan monohydrate (Sigma Aldrich Chemical Co., US) dissolved in sterile normal saline at a dose of 186,9 mg/kg body wt intraperitoneally. After three days, mice with blood glucose above 200 mg/dl were used for the experiment. Prior to initiation of this experiment, the animals were fasted for 8-12 h (Szkudelski, 2001), but allowed free access to water until the end of the experiment. [6] Experimental design: The mice were randomly divided into five experimental groups of six mice each. These groups included: the negative control group orally administered with 0,2 ml suspension 0,5% CMC-Sodium. Alloxan-induced diabetic control mice, orally administered with 0,2 ml of the reference drug glibenclamide (3mg per kg body weight, positive control grup) and alloxan induced diabetic experimental mice containing 10 mg/ 20g bw, 20 mg/ 20g bw and 40 mg/ 20g bw of plant dry extracts seeds of S. mahagoni Jacq. group 1. Diabetic mice given with dry extract seeds of S. mahagoni Jacq (10mg/20g bw), group 2. Diabetic mice given with dry extract seeds of S. mahagoni Jacq. (20mg/20g bw), group 3. Diabetic mice given with dry extract seeds of S. mahagoni Jacq. (40mg/20g bw). Blood sampling: Blood sampling was done by sterilizing the tail with 10% alcohol and then nipping the tail at the start of the experiment and this was repeated after 2nd, 4th, 6th and www.wjpr.net Vol 6, Issue 2, 2017. 1336
24th h, respectively. Every time the blood glucose levels were determined with a glucometer (EasyTouch GCU Meter). Data management and statistical analysis: Data was entered in the Microsoft Excel spread sheet, cleaned and then exported to Statistical Package for SocialSciences (SPSS Version 17.0) software for analysis. Results were expressed as Mean ± SEM. Differences in the means between the groups were statistically analyzed by One-way ANOVA followed by Bonferroni-Holm test for post hoc analysis with p<0.05 was considered significant. RESULTS AND DISCUSSION Result of the Swietenia mahagoni for thousand kilograms of each powdered plant material was extracted in 10 L etanol 96% for three times 24 h. The filtrat separated from solvent using a Buchner fannel. The filtrat was than evaporated using a rotary evaporator.the mahagony extract is added avicel : cab-osil with the ratio of 70:30 with the of comparisons dried 4:1. Tabel 1. Effects of oral administration of Dry Extracts of Swietenia mahagoni seeds in alloxan-induced diabetic BALB/c mice. Treatment CMC-Na 0,5% Glibenclamide ( 3mg/kg body weight ) Dry Extracts of Swietenia mahagoni seeds ( 10mg/20g body weight ) Dry Extracts of Swietenia mahagoni seeds ( 20mg/20g body weight ) Dry Extracts of Swietenia mahagoni seeds ( 40mg/20g body weight ) Blood glucose levels (mg/dl) 0 h 2 h 4 h 6 h 24 h 580,00 ± 559,33 ± 584,33 ± 590,67 ± 581,33 ± 12,283 13,966 9,992 9,333 6,858 542,00 ± 466,17 ± 325,50 ± 247,17 ± 333,50 ± 34,966 30,551 19,318 13,055 15,466 497,00 ± 413,67 ± 382,83 ± 327,83 ± 261,33 ± 13,621 31,571 37,921 36,151 41,240 507,83 ± 401,00 ± 345,83 ± 252,00 ± 186,67 ± 21,370 30,905 37,139 32,718 14,673 491,83 ± 403,17 ± 281,17 ± 170,17 ± 98,83 ± 26,962 35,745 25,527 27,952 4,126 The present study was carried out to evaluate the effect of dry extracts of Swietenia mahagoni seeds on experimentally induced diabetes mellitus in mice. The blood glucose lowering effect of dry extracts Swietenia mahagoni seeds was tested in positif control diabetic and experimentally induced diabetic mice. The dry extracts of Swietenia mahagoni seeds was given for one days 24 h in alloxan induced diabetic rats. Results were expressed as Mean±Standard Error of Mean (SEM) of five mice per group; Means followed by similar upper case letters in the same column are not significantly different at p 0.05 by ANOVA and post ANOVA (Bonferroni-Holm) test. www.wjpr.net Vol 6, Issue 2, 2017. 1337
As depiced in table 1, the dry extracts lowered blood glucose levels in a dose independent manner; however the 20 and 40 mg/20g body weight dry extracts doses significantly lowered blood glucose levels from the 2 h to the 24 h. Similarly, the dose 10 mg/20g body weight and positif control dose glibenclamide 3 mg/kg body weight lowered blood glucose levels no diferents. Oral administration of both the dry seeds extracts of Swietenia mahagoni at 10, 20 and 40 mg/20g body weeight doses to alloxan-induced diabetic BALB/c mice had significant effect on the blood glucose levels. The dry extracts of Swietenia mahagoni seeds showed potent antidiabetic activity in alloxaninduced diabetic mice. The blood glucose levels in animals were measured 72 hours after administration of alloxan which was done according to experiment of. [7] This dose range for mices aggres with the optimal and safe alloxan dose observated in this study of 186,9 mg/kg body weight. This loss is suggested to be due to kidney tubular cell necrotic toxicity at high doses of alloxan. [5] Induction of diabetes using alloxan-monohydrate to male BALB/c mice avails a functional and physiological model for the study of hypogycemic agents that could be used in the management of diabetes mellitus. In the present study, the rise in blood glucose level in experimental diabetic rats was also very high. The dose of Swietenia mahagoni (1000 mg/kg body weight), used in this study was selected in keeping conformity with the dose used in different research. [5] The observation that both of Swietenia mahagoni seeds at dose of 10, 20 and 40 mg/20g body weight lowered blood glucose from the second to the 24 h indicates that these extracts have antidiabetic constituents. These antidiabetic activity compound thought to generate dominant hypoglycemic activity in mahogany plants Swietenia mahagoni Jacq is swietenin. Swietenin refers to a class of tetranortriterpenoid compound representing natural agonist PPARγ (Peroxisome Proliferator Activated Receptor) with an action mechanism of activating the gen-responsive insulin which can stimulate insulin to form and translocate GLUT (glucosetransporter) to the cell membrane in peripheral organs so that the absorption and the use of peripheral glucose increased. [5,7] Another function of this PPARγ activation is enhancing the metabolism of carbohydrates and fats and improving insulin sensitivity. [7] The presence of dry extracts of Swietenia mahagoni seeds is an indication of the holistic nature of herbal medicine antidiabetic. www.wjpr.net Vol 6, Issue 2, 2017. 1338
CONCLUSION In conclusion, the findings of this study indicate that, the oral administration of dry extracts of Swietenia mahagoni seeds at doses of 10, 20 and 40 mg/20g body weight demonstrated antidiabetic activity in a dose independent manner comparable to that demonstrated by glibenclamide at 3 mg/kg body weight. The demonstrated antidiabetic activity could be associated with the phytonutrients present in Swietenia mahagoni seeds. The study recommends continued used dry extract Swietenia mahagoni seeds suitable to prepare formulation for treatment of diabetic. CONFLICT OF INTERESTS Declared None. ACKNOWLEDGMENT This reseacrh was financialy support by Hibah Kompetensi 2016 the Ministry Technology and Research and Higher Education, Indonesia. of REFERENCES 1. Khan AKA, Mahtab H, Grant J, Stewart M, Ahmed T, Haq JA, editors. 2007. Diabetes Mellitus 2 nd. Dhaka : Diabetic Association of Bangladesh. 2. Wild S, Roglic G, Green A, Sicree R, King H. 2004. Global Prevalence of Diabtes : Estimates for the year 2000 and projections for 2030 Diabetes Care; 27: 1047-53. 3. Bennett PN, Brown MJ, editors. 2008. Clinical Pharmacology. 10 th ed.churchill Livingstone, p 14-617. 4. Yelaware, P.N., Gunashekar, D.R., Faiyaz, A., Asna, U., 2014. Pharmacological Effects And Active Phytoconstituents of Swietenia mahagoni: A Review. International Journal of Pharmacy and Pharmaceutical Sciences Medicine. 12: 2. 5. Li, D., Chen, J., Chen, Q., Li, G., Chen, J., Yue, J.,Chen, M., Wang, X., Shen, J.,Shen, X., and Jiang, H., 2005. Swietenia mahagony Extract Shows Agonistic Activity to PPARγ and Gives Ameliorative Effects on Diabetic Mice. Acta Pharmacologia Sinica. (2): 220 222. 6. Szkudelski, T., 2001. The Mechanism of alloxan and streptozotocin action in beta-cells of the rat pancreas. Physiol. Res., 50: 536-546. 7. Etuk EU and Muhammad BJ. Evidence based analysis of chemical method of induction of diabetes mellitus in experimental rats. Int J Res Pharm Sci. 2010; 1(2): 139-42. www.wjpr.net Vol 6, Issue 2, 2017. 1339