Research & Reviews: Journal of Pharmacology and Toxicological Studies
Menue-ISSN: 2322-0139 p-ISSN: 2322-0120
e-ISSN: 2322-0139 p-ISSN: 2322-0120
Department of Pharmcology, Rakshpal Bahadur College of Pharmacy, Bareilly- 243001, Uttar Pradesh, India.
Received date: 03 March 2014; Revised Date: 27 April 2014; Accepted date: 04 May 2014
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We evaluated the hypoglycemic effects of methanol, water, petroleum ether, chloroform extract of Clitoria ternatea leaves. The hypoglycemic effect was evaluated in Streptozotocin induced diabetic rats for acute and sub acute effects. The extract of Clitoria ternatea also significantly (P<0.001) reduced blood glucose level in Streptozotocin induced diabetic rats twelve hours after administration.
Hypoglycemic, Clitoria ternatea, antidiabetic activity.
Diabetes mellitus (DM) is a widespread disorder, which has long been recognized in the history of medicine, before the advent of insulin and oral hypoglycemic drugs, the major form of treatment involved the use of paints. More than 400 plants are known to have been recommended ad recent investigations have affirmed the potential value of some of these treatments [1].
Diabetes mellitus is a group of endocrine syndromes characterized by hyperglycemia; altered metabolism of lipids, carbohydrates, and proteins, and an increased risk of complications from vascular disease. Most patients can be classified clinically as having either type I diabetes mellitus (type I DM formerly known as insulin dependent diabetes of IDDM) or type II diabetes mellitus (type II DM formerly known as non-insulin dependent diabetes of NIDDM) [2].
A good-looking perennial twining herb with terete stems and branches, leaves compound, imparipinnate, leaflets 5-7, sub- coriaceous, elliptic- oblong, obtuse; flowers blue or white , solitary axillary or in fascicles, corolla papilionacaous; fruits nearly straight, Flattened pods, Sharply beaked; seeds 6-10, smooth, yellowish brown. Clitoria ternatea is used as aphrodisiac tonic and are useful in ophthalmopathy. The leaves are useful in otalgia, hepatopathy and eruptions. The root also has anti-inflammatory, analgesic and antipyretic properties. Clitoria ternatea is used in leucoderma, burning sensation and pains. The roots are used as bitter, refrigerant, ophthalmic and laxative [3,4,5].
Plant material
The fresh leaves of Clitoria ternatea was collected during the month of September 2011, from the pratap Nursury, karamchari nagar Bareilly. The plant materials was taxonomically identified and authenticated by Dr. Umesh Chand Pandey, HOD and in charge Botany Department, Bareilly college, Bareilly (BCB/BOT/376/24-01-2012).
Preparation of Extract
The leaves of Clitoria ternatea were shaded dried until cracking sound was observed during breakage, and then these are made into coarsely powdered from using dry grinder. The powder leaves of the plant (500 gm) was macerated with each different solvents methanol, water, chloroform, petroleum ether (1500 ml) at room temperature for 72 hours with occasionally stirring. The extracts were separated from the residues by filtering 1st through several layers of muslin cloth for coarse filtration and then through what man No. 1 filter paper. The residue was further extracted using the same procedure. The filtrates obtained were combined and then evaporated to dryness at temperature not exceeding 400C and then give moderate heating on water bath at temp 40±50C. The extracts were kept indifferent Petri dish and it was stored in refrigerator (-4c) at cool place till use. During experiment the crude extracts were diluted (100 mg of the extract was dissolved in 0.5 ml water) with distilled water just before administration to the animal [6,7,8].
Animals
Male swiss albino mice of body 150-200 gm weight were taken before and after experiment with the help of single pan balance were used for the study. The animals were housed in clean metabolic cages and maintained in controlled temperature (27± 20C) and light cycle (12 hrs. light and 12 hrs. dark). They were fed with standard pellet diet (Gold mohar brand, Lipton India Ltd.) and water. The protocol was approved by Institutional animal ethics committee 1452/po/a/11/cpcsea.
Streptozotocin
Streptozotocin (STZ) is a naturally occurring nitrosourea product of Streptomyces achromogenes. Usually, the intraperitoneal injection of a single dose (25 mg/kg body weight) of it exerts direct toxicity on β cells resulting in necrosis within 48-72 h and causes a permanent hyperglycemia. STZ breaks nuclear DNA strand of the islet cells [9].
Preparation of Dose
The Dose of 200 mg/kg and 400 mg/kg of methanol extract was selected for the test. All the doses was given orally after making emulsion in vehicle i.e. 1% acacia gum and the standard drug i.e. glibenclamide was given orally (10 mg/70kg) in the vehicle.
Effect of different extract on streptozotocin induced diabeticrats
Induction of diabetes
Streptozotocin manufactured by Sisco Research laboratories Pvt. Ltd. Mumbai, India and was freshly dissolved in 0.1 M citrate buffer (pH = 4.5) at the dose of 25 mg/kg body weight and injected intraperitoneally within 15 min of dissolution in a vehicle volume of 0.4 ml with 1 ml of tuberculin syringe fitted with 24 gauge needle, where as normal control group was given citrate buffer only (0.4 ml). Diabetes was confirmed by the determination of fasting glucose concentration on the third day post administration of streptozotocin [10].
Sample collection
Blood sample were collected from tail nipping and glucose level was determined by an automatic electronic glucometer (Accuchek comfort).
Procedure
After checking the fasting blood glucose in overnight fasted diabetic rats. They were divided into five groups of five rats each and one group of non-diabetic rats.
All the doses were given in the following manner [3]
• 1st Group- normal control group received vehicle.
• 2nd Group-diabetic control received vehicle.
• 3rd Group-Received methanol extract at dose of 200 mg/Kg orally.
• 4th Group- Received methanol extract at dose of 400 mg/Kg. orally.
• 5thGroup-Received water extract at dose of200 mg/kg orally.
• 6th Group- Received water extract at the dose of 400 mg/kg orally.
• 7th Group-Received chloroform extract at the dose of 200mg/kg orally.
• 8th Group Received chloroform extract at the dose of400 mg/kg orally.
• 9th Group Received petroleum extract at the dose 200 mg/kg orally
• 10th Group Received petroleum extract at the dose 400 mg/kg orally
• 11th Group- Received standard drug i.e. Glibenclamide (10 mg /Kg. in Vehicle) orally.
The treatment was continued for 4 hours. During the period water was supplied ad libitum. All the doses were administered orally by the oral feeding needle. The effect of extract on Blood glucose levels was estimated on overnight fasted rats on 0 hour, 30 min, 60min, 120min and 240min by the method described before. The general behaviors of the animals were recorded. The blood glucose level in (Mean ± SEM) is shown in the Table.
Effect of different extract on oral glucose tolerance test
The hypoglycemic effect of extracts of Clitoria ternatea leaves was study on glucose loaded rats..
Table 1: The Antihyperglycemic effect of different extracts on Glucose Loaded rats
The effect of extracts on the glucose loaded animal the results shows that methanol extract is most potent than the any other extract. While chloroform extract shows minimum potency the blood glucose level. Petroleum ether extract is better than the chloroform extract but it shows less activity than water extract and methanol extract.
Graph 1: comparative effect of different extracts on glucose loaded rat
Table 2: The comparative Antihyperglycemic effect of different extracts on STZ induced Diabetic rats
Diabetic control shows highest blood glucose level .on the other hand standard drug shows the best result among the all extract. But methanol dose 400mg/kg shows prominent result. Chloroform shows minimum effect on the blood glucose level. In the 200mg/kg methanol also shows good effect but these was not good than the 400mg/kg of the methanol extract.
Graph 2: The comparative Antihyperglycemic effect of different extracts on STZ induced Diabetic rats
Table 3: The comparative sub-acute Antihyperglycemic effect of different extracts on STZ induced Diabetic rats
The result of the sub acute activity is much differs than the acute study this result shows that on the long term use of extract the dose 200mg/kg is much better to control the blood glucose level tan the 400mg/kg dose. After the 15 days the methanol extract of 200mg/kg dose shows minimum blood glucose level than other extract. In the all extract, on long term use 200mg/kg has a better result than the 400mg/kg. but the effect were same chloroform extract has minimum effect .petroleum ether extract is better effect than chloroform extract but poor effect than the methanol extract and water extract.
Graph 3: The comparative sub-acute Antihyperglycemic effect of different extracts on STZ induced Diabetic rats.
Table 4: The comparative sub-acute effect of different extracts on body weight of rats
Graph 4:
The effect of the standard drug and different extract the methanol extract of 200mg/kg dose showed 15 gm in the fifteen days and standard drug showed 10 gm weight gain in the animal. But in the case of chloroform extract there is something different because it showed 7gm weight loss in the 200mg/kg dose. Water 400mg/kg, 200mg/kg showed within 3gm ± weight difference in the animal. Petroleum ether 400mg/kg also showed 7gm weight loss in the animal during 15 days study. Diabetic control group showed 3gm weight gain in the animals.