Hypoglycemic effect of wheatgrass juice in Alloxan induced diabetic rats

About Authors:
M. R. N. Shaikh, Majaz Quazi, Dr. R. Y. Nandedkar
A. R. A. College of Pharmacy,
Nagaon,
Dhule – 06.

Abstract:
Wheatgrass has many characteristics desirable for health. It is known for its healing property. It is used as a cleansing and purifying agent and yet can be considered as a bodybuilder because of its nutritional value. It contains about 70% of chlorophyll which is very much comparable to alfalfa plant. Due to its high chlorophyll content it is also referred as Green Blood. Wheatgrass contain number of amino acids, vitamins, enzymes which contribute significantly to its health benefits. This article is nothing but an attempt to highlight one of the benefits of wheatgrass. The present investigation is undertaken in order to evaluate hypoglycemic property of wheatgrass juice.

Introduction:
Wheatgrass is commonly found herb in India and is known for its nutritional value. It is known as Triticumaestivum (T. aestivum) in botanical terms. This plant belongs to the Poaceae family. It contains enzymes like cytochrome, lipase, amylase, transhydrogenase, SOD (super oxide dismutase). Beside this, it contain all essential amino acids mainly alanine, aspartic acid, glutamic acid, arginine, serine which are helpful in providing sufficient amount of protein in the body ^[1]. Traditionally it is believed to be having anticancer, immunobodulatory, hypolipidaemic, antibacterial activities. In this communication, the hypoglycaemic action of wheatgrass juice in alloxan induced rats is reported ^[2].

Material and Methods
Growing of the grass and preparation of grass juice
The grass of T. aestivum used in this study was grown in the garden area of the institute until required for experiments. Overnight soaked T. aestivum seeds were then evenly spread over soil and further covered with soil. Small quantities of water were sprinkled evenly over soil and 3-4 hours indirect sunlight was allowed daily for growth of grass. On the tenth day, when grass is about 6″ tall, it is cut ½″ above the surface of soil. To harvest continuous supply of fresh grass, pots were similarly planted at one-day interval. Twenty grams of above harvested fresh grass was cut into further small pieces ^[4]. Then the juice was prepared by grinding the chopped grass in the juicer. The residue was twice resuspended in 3 ml of sterile water and similarly squeezed. Only fresh juice without any additives was used for the pharmacological evaluation.

Fresh T. aestivum grass was subjected to qualitative tests by standard methods as described by Harborne for preliminary phytochemical analysis ^[6].

Animals
Wistar rats of both sex and weighing between 150-200 g were used. The animals were housed under standard laboratory conditions, maintained on a natural light and dark cycle and had free access to food and water. Animals were acclimatized to laboratory conditions before the experimentation. All experiments were carried out between 9.00 a.m. and 15.00 p.m (throughout the study period). The experimental protocols were approved by the Institutional Ethics Committee and conducted according to the Indian National Science Academy Guidelines for the use and care of experimental animals.

Drugs
Alloxan (Loba chemie, Bombay), Gliclazide (Panacea Biotec), Glucose estimation kits (Span diagnostics Ltd., Surat, India) were used.

Evaluation of antihyperglycemic activity
Hyperglycemia was induced in the animals by single injection of alloxan monohydrate (150 mg/kg i.p.). It was confirmed after 48 hours (on third day). Animals (having blood glucose level greater than 225 mg/dl) were divided into different groups. Control group received no drug treatment, standard group received gliclazide (8 mg/kg, p.o) and test groups received test compound (50 & 100 mg/kg, p. o.) once a day up to 14th day of alloxan injection. Blood glucose was estimated in all the groups on 15th day ^[5]. Animals were sacrificed and their liver tissue was used for the estimation of glycogen.

Estimation of liver glycogen
Accurately weighed about 1.0 g of liver tissues, placed the tissues in calibrated centrifuge tube containing 2 ml of KOH (300g /l), and heated in a boiling water bath for 20 min. with occasional shaking. The tubes were cooled in ice, 0.2 ml of saturated sodium sulphate was added, and mixed thoroughly. Then glycogen was precipitated by adding 5 ml of ethanol and the precipitate was removed by centrifugation. The precipitates were dissolved in distilled water (10 ml) with gentle warming. One ml of this solution was added in duplicate in test tubes calibrated at 10 ml, to this 1 ml of HCl (1.2 mol/l) was added After placing marble on the top of each tube, heated in boiling water bath for 2 hours. After 2 hours 1 drop of phenol red indicator was added and neutralized with NaOH (0.5 mol/l). It was diluted to 5 ml with distilled water and the glucose content was determined. Glycogen content was expressed as g / g of liver tissue ^[7].

Statistical analysis
One specific group of rats was assigned to one specific drug treatment condition and each group comprised of six rats (n=6). The data was analyzed using one-way ANOVA followed by Dunnett’s test. In all the tests the criterion for the statistical significance was p<0.05.

Individual observations of Blood glucose level in mg/dl:

Individual observations of Liver Glycogen level in mcg/g: