IOSR Journal of Pharmacy and Biological Sciences (IOSR-JPBS) e-ISSN:2278-3008, p-ISSN:2319-7676. Volume 11, Issue 5 Ver. I (Sep. - Oct.2016), PP 17-26 www.iosrjournals.org Evaluation of Antidiabetic Activity of Ethanolic Extract of Trema Orientalis (L.) Blume Leaves. Jiji .K.N1*, Pramod.C2 , Boby S Prasad2, DR.P.Muralidharan1 1Department of Pharmacology, C.L.Baid Metha College of Pharmacy, Chennai. 2 University College of Pharmacy, Cheruvandoor, Kottayam, Kerala. Abstract: Context: Trema orientalis(L.)Blume is a species of flowering tree in the hemp family Cannabaceae. This common tree is widely used in African folk medicine for many diseases, for example asthma, cough, dysenteria and hypertension Objectives: The objective of the study was to evaluate the antidiabetic activity of ethanolic extract of Trema orientalis (L.) Blume leaves by in vivo methods. Material and methods: The leaves of Trema orientalis were dried under shade and then powdered, and extracted with ethanol by hot continuous extraction method using soxhlet apparatus. Preliminary phytochemical studies were carried out on extract. The antidiabetic potential of ethanolic extract of T.orientalis (EETO) was evaluated by using Streptozotocin-Nicotinamide induced type 2 diabetic rat model. Two doses of the EETO (250mg/kg and 500mg/kg) were administered to normal and experimental diabetic rats for 14 days. The blood samples were withdrawn from the retro orbital sinus on 1, 7 and 14th days of extract administration and fasting blood glucose levels were estimated. In addition, changes in body weight and serum lipid profiles, assessed in the extract treated diabetic rats were compared with diabetic control and normal animals. OGTT and histopathological studies of pancreas were also performed. Results and discussion: The study showed that, extract treatment in diabetic rats at two dose levels showed a decrease in fasting blood glucose levels. The total cholesterol and serum triglycerides levels were also reduced and the HDL cholesterol levels were increased upon treatment with the extract thus proving the potent antidiabetic property of the plant. Recovery of body weight of extract treated diabetic rats gave further evidence for dose related antidiabetic activity of EETO. The extract did not produce any significant effects on normal animals but significantly increases glucose tolerance of normal animals, which further confirms the safety and antidiabetogenic action of the extract. Conclusion:The study confirmed the potent antidiabetic activity of the plant Trema orientalis (L.) Blume. Keywords: Diabetes mellitus, Antidiabetic activity, Oral glucose tolerance test. I. Introduction Diabetes mellitus is a complex and a multifarious group of disorders that disturbs the metabolism of carbohydrates, fat and protein. It results from shortage or lack of insulin secretion or reduced sensitivity of the tissue to insulin [1]. Diabetes is a chronic medical condition, it can be controlled by medication and lifestyle modification throughout the life span. The clinical manifestation leading damage to the small blood vessels, referred to as microvascular disease. Diabetes is also an important factor in accerating the hardening and narrowing the arteries (atherosclerosis), leading to strokes, coronary heart disease, and other large blood vessel diseases. This is referred to as macrovascular diseases [2]. Currently there are over 150 million diabetics worldwide and this is likely to increase to 300 million or more by the year 2025 [3][4].Statistical projection about India suggest that the number of diabetics will rise from 15 million in 1995 to 57 million in the year 2025 making it the country with highest number of diabetics in the world [3]. This dreadful disease is found in all parts of world and is becoming a serious threat to mankind. A lot of synthetic chemical agents are available to control and treat diabetic patients, but total recovery from diabetes has not been reported till the date. Plants provide a potential source of hypoglycemic drugs and are widely used in several traditional system of medicine. The effects of these plants may delay the development of diabetic complications and correct the metabolic abnormalities by various mechanisms [5]. Currently available oral antihyperglycemic agents in clinical use have characteristic profile of side effects. Management of diabetes with agents devoid of any side effects is still a challenge to the medical system. This has led to an increase in the demand for natural products with antihyperglycemic activity having fewer side effects [6]. It is therefore imperative that the larger number of crude drugs which are in use as complementary and alternative medicine claiming to be useful in diabetes, be subjected to scientific evaluation. Traditional antidiabetic plants might provide a new oral hypoglycemic compounds, which can counter the high cost and poor availability of the current medicine present day drugs for many rural populations in developing countries [7][8]. DOI: 10.9790/3008-1105011726 www.iosrjournals.org 17 | Page Evaluation Of Antidiabetic Activity Of Ethanolic Extract Of Trema Orientalis (L.) Blume Leaves. Trema orientalis(L.)Blumeis a species of flowering tree in the hemp family Cannabaceae. This common tree is widely used in African folk medicine for many diseases, for example asthma, cough, dysenteria and hypertension [9].The root of this plant has been used for treatment of trauma, blood stasis, hematuria and bleeding of intestine and stomach [10][11].The root, bark and leaves are used in epilepsy. The stem bark is used as a poultice for muscular pain [12]. The study aims to scientifically evaluate whether the ethanolic leaf extract of T.orientalis has antidiabetic property. II. Materials And Methods 2.1Chemicals and regeants Streptozotocin obtained from Himedia (Mumbai, India). Nicotinamide and glibenclamide were obtained from (Alcon Pharmaceuticals, Mumbai). All other chemicals and reagents used were of analytical grade. 2.2. Plant materials The fresh leaves of T. orientaliswere collected fromKottayam M G University Campus on 5th April 2012. The plant was identified and authenticated from School of Environmental science, M.G University, Kottayam. A voucher specimen (SES.M.G.UTY NO. 1458) is preserved at the Herbarium of School of Environmental science, M.G University, Kottayam.Freshly collected plant materials were washedunder running tap water and distilled water to removeadhering dust and then dried under shade. The driedsamples were powdered in a Mechanical grinder and usedfor solvent extraction. 2.3. Solvent extraction The extraction of the dried powdered plant samples were carried out by hot continuous extraction method in Soxhlet extractor with ethanol as solvent for 10 h. The extract obtained was collected and concentrated by gentle heating followed by using vacuum evaporator. The concentrated extract was weighed and the percentage yield was calculated. Then the extract was subjected to Preliminary Phytochemical Evaluation. 2.4. Pharmacological evaluation 2.4.1Experimental animals Healthy adult Wistar male albino rats between 4 to 8 weeks of age and weighing about 150–250 g were used for the study. Animals were procured from the animal house of University College of Pharmacy, Cheruvandoor. Housed in polypropylene cages at a temperature of 25-30˚C and relative humidity 35 – 45%, light and dark cycles of 12 and 12 h respectively for one week before and during the experiments. Animals were provided with standard rodent pellet diet (Dayal Industries, Banglore) and the food was withdrawn 12-18 h before the experiment though water was allowed ad libitum. All studies were performed in accordance with the guide for the care and use of laboratory animals, and approved by the Institutional Animal Ethical Committee of UCP, Cheruvandoor, Kottayam, India (010/MPH/UCP/CVR/12). 2.4.2. Preparation of the test samples Extract as well as drug was suspended in 0.5% carboxymethyl cellulose (CMC) in distilled water prior to oral administration to the experimental animals. 2.4.3. Effect Of Ethanolic Extract Of T.orientalis Leaves In Normoglycemic Rats Effect of EETO in normoglycemic rats was evaluated by a method described by Shirwaikar et al[13].For normoglycaemic study, over night fasted animals were randomly divided into four groups of six anmals each. Group I - Normal Control rats received vehicle solution. Group II - Normal rats treated with EETO 250 mg/ kg body weight. Group III - Normal rats treated with EETO 500 mg/ kg body weight. Group IV - Normal rats treated with glibenclamide 0.6 mg/ kg body weight suspended in aqueous Solution [14]. The vehicles and the drug/extract were administered orally using intra gastric tube daily for 2 weeks. The blood samples were withdrawn from the retro orbital sinus on 1, 7 and 14th days of extract administration. The fasting blood glucose levels were estimated by glucose oxidase–peroxidase reactive strips and a glucometer(ONE TOUCH select simple, manufactured by Johnson’s & Johnson Ltd,India). DOI: 10.9790/3008-1105011726 www.iosrjournals.org 18 | Page Evaluation Of Antidiabetic Activity Of Ethanolic Extract Of Trema Orientalis (L.) Blume Leaves. 2.4.3. Oral Glucose Tolerance Test (OGTT) The oral glucose tolerance test was performed in overnight fasted (18 h) normal rats by a method described by Shirwaikar et al[15]. Rats divided into four groups, each group contains six animals. Group I - Normal Control rats received vehicle solution. Group II - Normal rats treated with glibenclamide 0.6 mg/ kg body weight suspended in aqueous Solution [14]. Group III - Normal rats treated with EETO 250 mg/ kg body weight. Group IV - Normal rats treated with EETO 500 mg/ kg body weight. Glucose (3 g/kg) was fed 30 min after the administration of extracts. Blood was withdrawn from the retro orbital sinus at 0, 30, 60, 90, and 180 min of glucose administration and glucose levels were estimated using glucose oxidase–peroxidase reactive strips and a glucometer(ONE TOUCH select simple, manufactured by Johnson’s & Johnson Ltd,India). 2.4.4. Effect of ethanolic extract of T.orientalis leaves in Streptozotocin-Nicotinamide Type2 diabetic rats[15].