D.Thaseena Begum et al. / Journal of Pharmacy Research 2011,4(6),1704-1706 Research Article Available online through ISSN: 0974-6943 http://jprsolutions.info Evaluating phytochemical and antidiabetic activity of ethanolic extract of whole o f indica Linn on alloxan induced diabetic Rats D.Thaseena Begum *1, K. Mahesh Kumar, P. Sravanthi, K. Sivasankara Prasad, K. Ravindrareddy 1Department of Pharmacology, P. Rami Reddy Memorial College of Pharmacy, Kadapa,Andhra Pradesh, . 516003 Received on: 11-02-2011; Revised on: 16-03-2011; Accepted on:21-04-2011 ABSTRACT mellitus is the most common endocrine disorder that impairs homeostasis resulting in severe diabetic complications including retinopathy, angiopathy, nephropathy, neuropathy and causing neurological disorders due to perturbation in utilization of glucose. The present study was designed to investigate the hypoglycemic properties of ethanolic extract of whole plant of Acalypha indica, which is widely used as a traditional treatment for diabetes mellitus. The present study was carried out to evaluate the antidiabetic activity of Acalypha indica ethanolic extract of whole plant of in alloxan induced diabetic rats for 7 days. The ethanolic extract at high dose (400 mg/kg) exhibited significant antihyperglycemic activity .The results showed that it has significant antihyperglycemic effect in experimental model of diabetes mellitus.

Key words: Diabetes, Hypoglycemic effect, Ethanolic extract of Acalypha indica , Alloxan.

INTRODUCTION Nature has endowed India with a unique gift of as many as 15,000 plant varieties. India and anti diabetic activity of the whole plant of Acalypha indica (L.) which is presented in accounts for two third of the flowering of the world of which about 2500 are reported the article. to possess medicinal and curative qualities1. About 70-80% of the populations depend on herbal remedies either partly or entirely. The export potential of herbal drugs has been MATERIALS AND METHODS estimated to be Rs. 2,840 million/annum for India and Rs.16, 000 million for China2. The India Council Medical Research (ICMR) has been doing research work with much Drugs and Chemicals: Ethanol (S.D. Fine chemicals Ltd, Mumbai), sodium hydroxide appreciable interest on herbal antidiabetic drugs. Inspite of the tremendous advancements (S.D. Fine chemicals Ltd, Mumbai), Sodium hydroxyl methyl cellulose (S.D. Fine made in the modern system of medicine for the management of diabetes mellitus, till data chemicals Ltd, Mumbai), Alloxan (sigma chemicals Co., U.S.A), Glibenclamide (Aristo on ideal or suitable drug is not available. pharma, Mumbai). Plant collection and Authentication The leaves of the plant Acalypha indica were collected from utukur, Kadapa District, Diabetes is a metabolic disorder characterized by symptoms like hyperglycemia, altered Andhra Pradesh. Authentified by Dr.K.Madhava Chetty, Department of Botany, Sri metabolism of lipids, carbohydrates and proteins. It also causes an increased risk of Venkateswara University, Tirupathi. complications from vascular diseases3.National Urban Diabetes Survey (NUDS) reveals that out of every eight Indians (above 20 years of age) living in urban metros is diabetic Preparation of the Extract and may be one in four either diabetes are potential candidate for diabetes inducement. The The Acalypha indica was collected, cleaned dried and powdered in a grinder –mixer to prevalence of diabetes in adults to be 13.2%. India has the highest diabetic population obtain a coarse powder and then passed through 40 mesh sieve. About 500 gm of powdered with an estimated 32 million people suffering from it. It is estimated that by 2005, India drug was extracted successively aqueous ethanol and water by soxhlet apparatus. The will have more than one fifth of the world diabetes population. The global diabetic extraction was carried out until the drug becomes exhausted. The solvents were recovered population is expected to double to 300 million in 2005 from 136 million in 1995. The from their extract by distillation under reduced pressure .The dried extract thus obtained increasing incidence in India during the period will be three fold from 19.4 million to 57.2 was kept in a desicator and was used for further experiments was well as used for million4.The ancient Ayurvedic drugs have mentioned about the permanent cure of diabetes identifying their chemical groups present .plant profile mentioned in the table no: 1 mellitus through restoration of to normal functioning5. Though this theory appears to be very sound as yet no such drug has been clinically investigated so far which Experimental Animals can completely bring back pancreas to normal functioning in diabetes. The therapeutic Male wistar rats (150-200 g) body weight were obtained from Raghavendra enterprises, objective of the modern system of medicine is to provide metabolic relief by bringing Bangalore, and they were housed under standard husbandry conditions, 25±50C temperature, down the blood glucose level to normal. This helps in the prevention, postponement and 12 h light/dark cycle with standard rat feed (Pravan agro Ltd. India) with water ad libitum. amelioration of the complications of diabetes mellitus and also in improving the quality The experimental protocol was approved by the Institutional Animal Ethical Committee of life. of P.Rami Reddy Memorial Collage of Pharmacy (1423/Po/11/CPCSEA/104/2011).Ethical

Acalypha indica linn an annual erect herb found throughout various parts of India, Table..1 The Acalypha indica plant scientific profile mentioned in the below table: Bangladesh, Srilanka, Philippines and tropical Africa. The plant is commonly known as Kingdom Plantae Indian acalypha indica and belongs to the family . The plant has wide uses Division Magnoliophyta in the traditional medicines of various countries and reported to posses diuretic, purgative Class Magnoliopsida and anthelmintic properties , besides being also used bronchitis, asthma ,pneumonia, Order 6 Family Euphorbiaceae scabies and other cutaneous diseases . Genus Acalypha Species Indica. Acalypha indica whole plant contain acalphine which is used in the treatment of sore Botanical name Acalypha indica. gums and have post antifertility effect7, anti venum properties 8, wound healing effects9, antioxidant effects10, anti inflammatory effects 11, acardial effects 12, diuretic effects 13, anti clearance for performing the experiments on animals was obtained from Institutional bacterial activites14, antihelminitic 15. The plant Acalypha indica (L.) was selected for the Animal Ethics Committee (IAEC). present investigation. Ayurvedic knowledge supported by modern science was necessary to isolate characterize and standardize the active constituents from herbal sources for anti Acute toxicity studies diabetic activity. Since, all parts of investigated plant were being used for various human Acute toxicity studies for aqueous ethanolic extract of Acalypha indica was carried out ailments and very less scientific work has been performed. The present investigation was in rats at different doses (1000 - 2000 mg/kg, orally), showed no gross evidence of any carried out to expose the chemical and therapeutically potential by evaluating phytochemical abnormalities in the mice up to the end of 72 hr of the observation period. This indicates the safety of extract. Further, it is reported 21 that ethanolic extract of Acalypha indica is found to be safe at a dose of 3000 mg/kg. Hence, further pharmacological investigation was *Corresponding author. carried at dose levels of 200, 400 mg/kg. Acute toxicity study was done as per OECD, D.Thaseena Begum 2006 Guidelines (OECD, 2006 Guidelines). M.pharmacy, Induction of diabetes Department of pharmacology, Alloxan (2, 4, 5, 6-tetraoxypyrimidine; 2, 4, 5, 6- tetrone) is an oxygenated P.Rami Reddy Memorial College of Pharmacy, pyrimidine derivative 16 and was originally isolated in 1818 by Brugnatelli and got its Kadapa, Andhra Pradesh, India-516003 name in 1838 by Friedrich Wohler and 17. Alloxan is a toxic glucose analogue, which selectively destroys -producing cells in the pancreas when

Journal of Pharmacy Research Vol.4.Issue 6. June 2011 1704-1706 D.Thaseena Begum et al. / Journal of Pharmacy Research 2011,4(6),1704-1706 administered to rodents and many Table no: 4 Effect of Aqueous ethanolic extract of Acalypha indica on body weight in alloxan other animal species. This causes an -induced diabetic rats. insulin-dependent diabetes mellitus Group Body weight (g) (called “Alloxan Diabetes”) in these Day 1 Day 7 animals, with characteristics similar I. Normal control 225.45±0.76 236±1.36 to type 1 diabetes in humans. Alloxan II. Diabetic control 220.91±0.82 200.7±0.6 monohydrate was obtained from S.D. III. Standard (10mg/kg) 205.32±0.67 196.5±1.57** Fine, Mumbai and all the other IV. ELAI (100mg/kg) 209.54±0.65 190.42±0.82* chemicals used were of analytical Values are given as mean±S.E.M. from six rats in each group. * Represents statistical signifi- grade and were acquired from cance vs. diabetic control (P < 0.05). ** Represents statistical significance vs. diabetic control commercial sources. (P < 0.01). Oral glucose tolerance test (OGTT) Hypoglycemic Effect of methanolic extract The oral glucose tolerance test was The results from the study clearly indicated that the ethanolic extract exhibited significant performed in overnight fasted normal hypoglycemic activity in alloxan -diabetic rats. The standard drug glibenclamide also animals. Rats divided into three Figure no:1 indicated a significant decrease of blood glucose levels. (Table 5) groups (n = 6) were administered distilled water 10 ml/kg, Aqueous ethanolic extract of Table no: 5 Effect of Aqueous ethanolic extract of Acalypha indica on fasting blood glucose Acalypha indica (AEEAI) 100 ml/kg p.o and glibenclamide (GBC) 10 mg/kg, respectively. level in diabetic rats. Glucose (2.5 g/kg) was fed 30 min after the administration of extracts. Blood was Group Fasting blood glucose concentration (mg/dl) withdrawn from retro orbital just prior to the drug administration (normal fasting) and at Day 1 Day 3 Day 5 Day 7 0, 30, 60 and 120 min of glucose loading. Blood glucose level was measured immediately I. Normal control 110.27 ±11.67 115.16 ±8.26 117.31 ±14.76 119.83±15.02 by using glucose kit (God – Pod Method). II.Diabetic control 309.45 ±29.35 318.87±35.67 326.4 ± 37.54 296.5 ±45.56 Alloxan (150 mg/kg S.C) Evaluation of antidiabetic activity III.Standard (10mg/kg P.O) 346.45± 27.46** 258± 40.34*** 192.45±38.67*** 142.7± 25.06*** Rats were divided into five groups of six rats (n = 6) each. Groups 1 and 2 served as IV. ELAI(200mg/kg) 339.67±21.86** 247.69±27.12*** 200.56±29.67*** 186.65± 23.82*** V. ELAI (400mg/kg) 330.45±16.54** 252.83±37.70*** 182.67±27.88*** 153.83± 20.93*** control and diabetic untreated control respectively. Group 3 served as standard and was treated with 10 mg/kg/day glibenclamide. Group 4 and 5 was treated with the Aqueous Values are given as mean±S.E.M. from six rats in each group. ** Represents statistical significance ethanolic extract of Acalypha indica (200 mg/kg, 400mg/kg p.o. for 7 days. Blood vs. control (P < 0.01). *** Represents statistical significance vs. control (P < 0.001). glucose levels were measured on day 1, 3, 5 and 7 of the study. DISCUSSION The fundamental mechanism underlying hyperglycemia involves over-production (excessive The various groups used in experiment hepatic glycogenolysis and gluconeogenesis) and decreased utilization of glucose by the Group 1 - Served as normal control and received 1% SCMC 2ml/kg per orally. tissues15. Persistent hyperglycemia, the common characteristic of diabetes can cause Group 2 – Alloxan 150 mg/kg (S.C) and received 1% SCMC 2ml/kg (P.O). most diabetic complications. In all patients, treatment should aim to lower blood glucose Group 3 - Alloxan 150 mg/kg + Glibenclamide (10 mg/kg, p.o.). to nearnormal levels. In our investigation, the normal glycaemic and oral glucose tolerance Group 4 - Alloxan 150 mg/kg + AEEAI (200 mg/kg, p.o.) test studies revealed that the ethanolic extract of whole plant of Acalypha Indica has the Group 5 - Alloxan 150 mg/kg + AEEAI (400 mg/kg, p.o.) capacity to lower blood glucose levels. AEEAI — Aqueous ethanolic extract of Acalypha indica SCMC— Sodium carboxy methyl cellulose The diabetic syndrome in rats administered alloxan is characterized by stable moderate hyperglycemia, glucose intolerance and altered but significant glucose stimulated insulin Statistical Analysis secretion16. In normal glycaemic study, the data indicates that the AEEAI treatment All the values of body weight, fasting blood sugar, were expressed as mean ± standard significantly reduced the blood glucose levels (***P<0.001) in the diabetic rats towards error of mean (S.E.M.) and analyzed for ANOVA and post hoc Dunnet’s t test. Differences the normal level in the 7 days of study period. between groups were considered significant at P < 0.05 levels. RESULTS Table .2 Qualitative chemical tests for In oral glucose tolerance test, treatment of AEEAI significantly reduced the serum glucose phyto constituents in Acalypha indica Linn level (***P<0.001) over the period of 120 min compared to normal control group. These EXTRACTION Tests Aqueous ethanolic extract data suggested that treatment with AEEAI showed tolerance to exogenously administration The dried powder of Acalypha Indica (50 gms) of Acalypha indica glucose in the treated group of rats in OGTT. was extracted with 80% aqueous ethanol by Alkaloids + soxhelation. The yield of aqueous ethanolic Carbohydrates + The characteristic loss of body weight associated with alloxan induced diabetes is due to extract is 9.46g. Flavonoids + increased muscle wasting in diabetes. The Acalypha Indica ethanolic extract treated Triterpenoids - animals recovered the body weight significantly towards normal level (*P<0.05). This Protiens + Phytochemical tests Resins — may be directly due to the lipid lowering activity of the extract or indirectly to the The phytochemical tests revealed the presence Sapoinins + influence on various lipid regulation systems. Steroids + of Alkaloids, proteins, flavanoids, reducing Tannins — sugars, gums and mucilage, saponins in volatile oils — CONCLUSION ethanolic extract. Mentioned table no: 2 lignin’s __ From the phytochemical screening major constituents present in the compounds are flavanoids and alkaloids. The phytochemical screening has given the basic foundation in Oral glucose tolerance test which the class of compounds possesses the anti diabetic activity. Administration of glucose (2.5 g/kg,) produces significant change in blood glucose level of normal rats. Treatment with AEEAI (100 mg/kg, p.o.) and GLB (10 mg/kg, p.o.) Aqueous ethanolic extract of Acalypha indica exhibit significant anti-hyperglycemic significantly reduced serum glucose level at normal fasting, at 0 minute, 30 minute, 60 activities in alloxan -induced rats. The Aqueous ethanolic extract of Acalypha indica also minute and 120minute compared to normal control group (Table 3). showed improvement body weight, OGTT, glucose as well as regeneration of Table no: 3 Effect of Aqueous ethanolic extract of Acalypha indica on OGTT in normal rats. pancreatic islets of langerhans so might be value in treatment of diabetes. However, further phyto pharmacological investigations are needed to identify the lead molecule and to Groups Blood glucose concentration (mg/dl) elucidate its exact mechanism of action for anti-diabetic effect. Normal 0 min 30 min 60 min 120 min. Fasting ACKNOWLEDGEMENT I. Glucose 121.34±6.12 252.45±4.34 284.23±5.27 236.87±4.38 169.57±3.22 We are thankful to Management of P. Rami Reddy memorial College of Pharmacy, (2.5g/kg) Kadapa, A.P for the providing the all facilities for carried out this work. II.GBC 102.21±2.26** 196.32±5.18** 225.58±2.65** 152.40±3.12** 132.36±2.02** (10 mg/kg) III.AEEAI 115.26±4.12** 201.43±6.98** 251.45±6.22** 186.52±7.02** 148.45±5.32** REFRENCES (100mg/kg) 1. Rajan. T.P.S., Chemical weekly, XL1994, (1), 51. 2. Sanjay Garg., CRIPS Vol.2 No.1, 2001, 16. Values are given as mean±S.E.M. from six rats in each group. ** Represents statistical signifi- 3. Alfred. G.G., Theodore. W.R., Aklaw. S.N. and Palmer. T., Goodman and Gillman, The cance vs. control (P < 0.001) Pharmacological Basis of Therapeutics, 8th Edn, Pergamon Press, New York, 1991, 147. 4. Express Pharma Pulse Vol.7, No.47, (2001). Changes in body weight 5. Dhar. M.L., Dhar. M.M., Dhawan. B.N., Mehtotra. B.N. and Ray. C., Indian journal of Experimental At the end of 7 days treatment the body weight of diabetic control group decreased whereas Biology, 1968, 6(4), 272. 6. 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Journal of Pharmacy Research Vol.4.Issue 6. June 2011 1704-1706