Objective: As per the ethnopharmacological information has Butea monosperma been used to treat diabetes mellitus by the tribal people of tropical and subtropical areas. However, there is no much more scientific report available about the antidiabetic property of the leaves of the plant. Hence, the study was undertaken to evaluate the antidiabetic effect of ethanolic extract of B. monosperma on blood levels of adrenaline-induced and glucose feed diabetic rabbits.Methods: The three different doses of the extracts (100, 200, and 400 mg/kg) were administered orally to an experimental animal. The animals were induced diabetics by adrenaline and high glucose diet. Blood glucose level was measured accordingly. For antidiabetic activity, photocolorimeter was used to monitoring the blood glucose level with crest kit box (GOP-POD method).Results: The extracts showed considerable dose-dependent activity. However, the dose 400 mg/kg showed considerable lower of blood glucose level. p&lt;0.01 indicates the significance result. 8 hrs reading 182.5Â±3.83 for 400 mg does is most effective for reducing blood sugar.Conclusion: The study indicates that the ethanolic extract of B. monosperma leaves possesses antidiabetic properties which suggest the presence of biologically active components

Behera, Pradipta Kumar

Mahapatra, Santosh Kumar

Panigrahi, Ashok K

Ray, Biswaranjan

Innovare Academic Sciences: E-Journals

Vol 10, Issue 12, 2017Online - 2455-3891 Print - 0974-2441EVALUATION OF ANTIHYPERGLYCAEMIC EFFECT OF BUTEA MONOSPERMA LEAF EXTRACT ON ADRENALIN INDUCED AND HIGH GLUCOSE FEED ANIMAL MODELBISWARANJAN RAY1*, SANTOSH KUMAR MAHAPATRA2, PRADIPTA KUMAR BEHERA1, ASHOK K PANIGRAHI31Department of PhD, Sambalpur University, Sambalpur, Odisha, India. 2Department of Pharmacy, Gayatri College of Pharmacy, Sambalpur, Odisha, India. 3Department of Pharmacology respectinely, Veer Surenrdra Sai Institute of Medical Sciences and Research, Sambalpur, Odisha, India. Email: crabiswa@gmail.comReceived: 22 July 2017, Revised and Accepted: 08 September 2017ABSTRACTObjective: As per the ethnopharmacological information has Butea monosperma been used to treat diabetes mellitus by the tribal people of tropical and subtropical areas. However, there is no much more scientific report available about the antidiabetic property of the leaves of the plant. Hence, the study was undertaken to evaluate the antidiabetic effect of ethanolic extract of B. monosperma on blood levels of adrenaline-induced and glucose feed diabetic rabbits.Methods: The three different doses of the extracts (100, 200, and 400 mg/kg) were administered orally to an experimental animal. The animals were induced diabetics by adrenaline and high glucose diet. Blood glucose level was measured accordingly. For antidiabetic activity, photocolorimeter was used to monitoring the blood glucose level with crest kit box (GOP-POD method).Results: The extracts showed considerable dose-dependent activity. However, the dose 400 mg/kg showed considerable lower of blood glucose level. p<0.01 indicates the significance result. 8 hrs reading 182.5±3.83 for 400 mg does is most effective for reducing blood sugar.Conclusion: The study indicates that the ethanolic extract of B. monosperma leaves possesses antidiabetic properties which suggest the presence of biologically active components.Keywords: Glucose, Adrenaline, Leaf extract, Hyperglycemia.INTRODUCTIONDiabetes mellitus is a chronic metabolic disorder characterized by high glucose concentration (hyperglycemias) due to impaired insulin secretion and or insulin resistance [1]. It is a metabolic disorder in which there is an inability to metabolize carbohydrate due to disturbances in insulin function and secretion. Diabetes is one of the oldest known diseases of the man whose devastating effect is increasing by the day and severity almost at an epidemic level. The identification parameter and treatment protocol are becoming complicated day-by-day. Diabetes is always associated with many other complications which may or may not be preexisting. The number of people suffering from the disease worldwide is increasing at an alarming rate with a projected 366 million peoples likely to be diabetic by the year 2030 as against 191 million estimated in 2000 (Wild et al., 2004). Developing countries are the most affected because of expansive and inadequate treatments (djr olo etd, 1998), coupled with the side effect associated with these drug [2].Thus the search of a new drug with low cost, more potential and without adverse effect becomes inevitable. A great number of medicinal plants have been used in the treatment of diabetes in a different part of the world. Many herbal products are the mixture of different plant extract used for a therapeutic purpose which is based on its ethnobotanical information. Many times plant product are subjected to animal screening but sometimes these product used by following traditional information on the basis of traditional use. Evaluation of the antidiabetic potentials of these plants becomes necessary to provide scientific proof and justify their uses in ethnomedicine.Butea monosperma is a species of Butea native to tropical and subtropical parts of the Indian Subcontinent and Southeast Asia, ranging across India, Bangladesh, Nepal, Pakistan, Sri Lanka, Myanmar, Thailand, Laos, Cambodia, Vietnam, Malaysia, and western Indonesia 4,5. In this study, the different doses of leaf extracts were screened for antidiabetic study [3].METHODSCollection and leaf extract preparationThe plant leaves were collected based on local ethnobotanical information. The collected plant materials were subjected to authenticated identification using the standard of nomenclature and classification. The authentification has been certified by the Scientist, Botanical Survey of India (BSI), Kolkata.The leaves were dried in the shade and powdered to get a coarse powder. About 800 g of dry, coarse powder was subjected to extraction with ethanol using Soxhlet apparatus. The residue of the extract obtained evaporated to dried mass and then the residue of the extract obtained was taken for the experiment [4,5].Chemicals and reagent usesAll chemicals and drugs were obtained commercially and were of analytical grade. All the reagent kits were obtained from crest India Ltd.Instrument usedFor antidiabetic activity photocolorimeter was used to monitoring the blood glucose level with crest kit box (GOP-POD method).Phytochemical screeningThe ethanolic extract of the leaves obtained was subjected to preliminary phytochemical screening to identify the chemical constituents. The methods of analysis employed were those described by Trease and Evans (1989) [6].© 2017 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons. org/licenses/by/4. 0/) DOI: http://dx.doi.org/10.22159/ajpcr.2017.v10i12.21501Research Article302Asian J Pharm Clin Res, Vol 10, Issue 12, 2017, 301-304 Ray et al. The presence of various constituent has been screened in a different chemical test using different chemical reagent. The finding of different chemical constituents is evidence for different pharmacological activity [7].Dose selectionThe three test drug dose and standard drug dose were selected by acute toxicity study (LD50).Animals and induction of diabetes mellitusHealthy albino rabbits of either sex, inbred in the departmental animal house, weighing between 2 and 2.5 kg, aged between 6 months and 1 year were used for this study. The animals were kept properly and maintain required condition suitable for experiment before and during experiment. Adrenaline diabetes can be produced by intravenous, intramuscular, subcutaneous, administration of different doses. In this study, adrenaline 8 mg/kg by intravenous route has been sued as the route is least toxic. The protocol of the study was approved by IAEC, Gayatri College of Pharmacy, Sambalpur, India, bearing Regd. No1339/ac/10/CPCSEA.Experimental designMethod used by Mohan et al. (1978) was followed to study the affect of BMLE on the fasting plasma glucose (FPG) at different hour’s intervals. Adrenaline was administered in the dose 0.8 mg/kg for induction of diabetics. In addition, high carbohydrate diet has been employed for experimental animal to maintain the good diabetic model. Adrenaline is responsible for hyperglycemia in an animal model, and high carbohydrate diet plays a synergistic role for maintaining diabetes [8,9]. The diabetic rabbits with FPG level 200-250 mg/dl were divided into different groups of six animals each in the following manner. The detail is present in Table 1.Determination of blood glucose levelsBlood samples from the overnight fasted diabetic rabbits were collected. Then, the drug and the vehicles in their indicated doses were administered orally to the respective groups. The effect of BMLE on FPG was compared to that of their respective vehicle and glibenclamide, the standard antihyperglycemia drug at different time interval by photocolorimeter. The detail is present in respective table.Statistical analysisBlood glucose levels were expressed in mg/dl as mean ± SEM. The data were statistically analyzed using ANOVA with multiple comparisons versus control group. The values of p<0.01 were considered as significant [10].RESULT AND DISCUSSIONAdrenaline-induced hyperglycemia has been described as a useful experimental model to study the activity of antidiabetic agents as adrenaline is responsible for elevated glucose label by producing glycogenolysis. The following table shows the result of the effect of three doses (100, 200, and 400 mg/kg) of the ethanolic extract of B. monosperma leaves.BM leaf extract administered in doses of 100, 200, and 400 mg/dl to three different group of diabetic rabbits whose FPG was estimated just before (Ohr) and 2, 4 as well as 8 hrs after the drug administration. In Table 2, the effect of vehicle on FPG has been noted. This table bears the report of the FPG of the diabetic rabbits 0 hr (before) and 2, 4 and 8 hrs intervals after administrations of different vehicles. Here, p<0.05 after analysis by one-way ANOVA. There is no significant difference between the mean FPG of all groups. Hence, none of the vehicles have any significant action. In Table 3, the effect of different doses of BM leaf extract on adrenaline-induced diabetic animal has been mentioned. BM leaf extract at 100, 200, and 400 mg/kg dose has been administered and glucose level has been monitored at 0 hr (before) and 2 hrs, 4 hrs and 8 hrs intervals after administration. One-way ANOVA analysis shows p<0.01 which indicates statistical significance. At 4 hrs reading and 8 hrs reading there is a significant lowering of FPG. In 8 hrs reading the lowering of FPG is very prominent. Table 4 is a consolidated table showing the effect of BMLE at 3 dose level and glibenclamide 0.5 mg/kg on mean FPG of different groups of rabbits as compared to their respective vehicles at 0, 2, 4, and 8 hrs interval of drug administration. Statistical analysis by one-way ANOVA followed by paired t-test indicates the significant of result which has been mentioned in the respective table. The highly significant result has been indicated as triple star within the table. BMLE 100 mg dose at 8-hour interval achieve 203.5±3.92.200 mg dose at 8 hrs interval achieve 191.33±5.14.Which is very effective and 400 mg achieve 182.5±3.83 which is very effective for reducing blood glucose level in the experimental animal.The peak effect with each dose was observed at 8 hrs with low dose the onset of antihyperglycemic action was delayed whereas with high dose an early response was observed [11,12].Table 2: Effect of vehicles on fasting plasma glucose in adrenaline-induced diabetic rabbits (acute study)S. No. FPG (mg/dl)Distilled water 2% gum acacia 5% Tween 800 hrs 2 hrs 4 hrs 8 hrs 0 hrs 2 hrs 4 hrs 8 hrs 0 hrs 2 hrs 4 hrs 8 hrs1 239 239 231 224 236 232 228 219 245 241 243 2392 237 234 238 225 238 234 236 238 235 236 231 2253 232 227 223 232 231 224 223 225 232 229 233 2244 244 228 237 238 243 238 237 231 234 233 235 2315 231 228 231 226 230 226 229 227 219 215 221 2136 226 233 228 226 225 228 227 223 227 229 224 233Mean 234.83 231.5 231.33 228.5 233.8 230.3 230.0 227.17 230.0 230.5 231.2 227.5One-way ANOVA p>0.05ns, ns: Not significant. This table bears the report of the FPG of the diabetic rabbits 0 hr (before) and 2, 4 and 8 hrs intervals after administrations of different vehicles. There is no significant difference between the mean FPG of all groups. Hence, none of the vehicles have any significant action on FPG levels of diabetic rabbits, and there is no significant difference among the effects of these three vehiclesTable 1: Details of Drug, Vehicle and Routes of AdministrationDrug/vehicle Dose per kg body weight Route of administration Nature of use5% Tween 80 2 ml Oral Vehicle for SRLEGlibenclamide 0.5 mg Oral Standard antihyperglycemic drugSRLE 100, 200 and 400 mg Oral Indigenous test drugAdrenaline 0.8 mg I.V Diabetes inducing agent303Asian J Pharm Clin Res, Vol 10, Issue 12, 2017, 301-304 Ray et al. Table 3: Effect of BMLE on FPG in adrenaline-induced diabetic rabbits (acute study)S. No.FPG (mg/dl)0 hr2 hrs4 hrs8 hrsT-80B-100B-200B-400GLT-80B-100B-200B-400GLT-80B-100B-200B-400GLT-80B-100B-200B-400GL123222423823923622921922922721323320820520416222419819718617122342352252432282332342212302092352242012171562312091881921623227238243233239229221228219213224210219201164233201210184169424523921922824124123720821521524322819517817323921817217817552352192332452212362152212311962312022012151502251901941891576219233228221227215229225209201221221199182176213205187166183Mean232.0231.33231.0234.8232.0230.5225.8222.0221.8207.8231.2215.5203.3199.5163.5227.5203.5191.33182.5169.5SEM3.543.33.592.783.23.613.603.123.643.123.234.193.406.684.033.673.925.143.833.78One-way ANOVA p>0.05ns. *p<0.01 statistically significant this table bears the report of the fasting plasma glucose of the diabetic rabbits. 0 hr (before) and 2 hrs, 4 hrs and 8 hrs intervals after administration of BMLE at 100, 200, and 400 mg/kg and gl at doses 0.5 mg/kg. There is a significant difference among the mean FPG value of different groups in each set at 2, 4 as well as 8 hrs interval. Result of post ANOVA tests is represented in next page, tableTable 4: Effect of BMLE on mean FPG in diabetic rabbits (acute study)GroupTreatmentDose per kg Bd. Wt.Mean FPG ± SEM (mg/dl)0 hr2 hr4 hr8 hrIGlibenclamide0.5 mg232.0±3.2207.8±3.12***163.5±4.03***169.5±3.78***I i5%tween802 ml232.0±3.4230.5±3.61231.2±3.23227.5±3.67IiiBMLE100 mg231.33±3.3225.8±3.60215.5±4.19203.5±3.92**IvBMLE200 mg231.0±3.59222.0±3.12203.3±3.40**191.33±5.14***vBMLE400 mg234.8±2.78221.8±3.64**199.5±6.68***182.5±3.83***This is a consolidated table showing the effect of BMLE at 3 dose level and glibenclamide 0.5 mg/kg on mean FPG of different groups of rabbits as compared to their respective vehicles at 0, 2, 4, and 8 hrs interval of drug administration. 8 hrs reading 182.5±3.83 for 400 mg does is most effective for reducing blood sugar Post ANOVA paired t-test  Test drug Vs. glibenclamide**Statistically significant**Statistically significant  Test drug /glibenclamide Vs. Vehicle***Highly significant304Asian J Pharm Clin Res, Vol 10, Issue 12, 2017, 301-304 Ray et al. CONCLUSIONThe study indicates that the ethanolic extract of B. monosperma leaves possesses antidiabetic properties which suggest the presence of biologically active components. The extract might be promoting glucose uptake and metabolism or inhibiting hepatic gluconeogenesis. BM leaf extract administered in doses of 100, 200, and 400 mg/dl to three different group of diabetic rabbits whose FPG was estimated just before (0 hr) and 2, 4 as well as 8 hrs after the drug administration. The results of this study depicted in Table 2 and Graph 1 reveal that the drug exhibited a significant effect on FPG of diabetic rabbits. Post ANOVA analysis of the data by paired t-test reveals that BMLE with 100 mg/kg dose reduced mean FPG levels in diabetic rabbits significantly at 8 hrs; with 200 mg/kg at 4 and 8 hrs; and with 400 mg at 2, 4 as well as at 8 hrs intervals in comparison to its vehicle. The peak effect with each dose was observed at 8 hrs. With low dose, the onset of antihyperglycemic action was delayed whereas with high dose an early response was observed [13].REFERENCES1. Ganong WF. Review of Medical Physiology. 21st ed. New York: McGraw-Hill Professional; 2003.2. Garatini S, Paoleti R, Bizzi L, Gmssi E, Vertua A. Drugs Affecting Lipid Metabolism. Amsterdam: Elsvier; 1961. p. 1-44.3. Kirtikar KR, Basu BD. Indian Medicinal Plants. Vol. 1. Allahabad, India: Lalit Mohan Basu, Prakashan; 1991. p. 785-9.4. Rosenthaler L. The Chemical Investigation of Plant. 1st ed. London: Bell and Sons; 1930.5. Suganya G, Kumar PS, Dheeba B, Sivakumar R. In vitro anti diabetic, antioxidant and anti-inflammatory activity of Clitoria ternatea. Int J Pharm Pharm Sci 2014;6(7):342-7.6. Ali M. Textbook of pharmacognosy. Treaseand Evans Pharmacognosy. 13th ed. London: Bailliere Tindall; 2002.7. Agbafor KN, Nwachukwu N. Phytochemical analysis and antioxidant property of leaf extracts of Vitex doniana and Mucuna pruriens. Biochem Res Int 2011;2011:459839.8. Kokate CK. Practical Pharmacognosy. 4th ed. New Delhi, India: Vallabh Publications, Prakashan; 1994.9. Naik SR, Barbosa Filho JM, Dhuley JN, Deshmukh V. Probable mechanism of hypoglycemic activity of bassic acid, a natural product isolated from Bumelia sartorum. J Ethnopharmacol 1991;33(1-2):37-44.10. Mandlik RV, Desai SK, Naik SR, Sharma G, Kohli RK. Antidiabetic activity of a polyherbal formulation (DRF/AY/5001). Indian J Exp Biol 2008;46(8):599-606.11. Evangeline RM, Murugan N, Kumar PP, Christhudas IV. In vitro studies on a glucosidase inhibition anti oxidant and free radical scavenging properties of Tecoma stans L. Int J Pharm Sci 2015;7(7):44-9.12. Ghosh MN. Statistical methods. In: Fundamentals of Experimental Pharmacology. 3rd ed. Kolkata: S.K. Ghosh and Others, Hilton and Company; 2005. p. 209-29.13. Grover JK, Vats V, Rathi SS. Anti-hyperglycemic effect of Eugenia jambolana and Tinospora cordifolia in experimental diabetes and their effects on key metabolic enzymes involved in carbohydrate metabolism. J Ethnopharmacol 2000;73(3):461-70.Graph 1: Effect of Bmle on Fpg in Diabetic Rabbit

EVALUATION OF ANTIHYPERGLYCAEMIC EFFECT OF BUTEA MONOSPERMA LEAF EXTRACT ON ADRENALIN INDUCED AND HIGH GLUCOSE FEED ANIMAL MODEL

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