EVALUATION OF WHEATGRASS JUICE FOR

ANTIDIABETIC ACTIVITY IN RATS

SYNOPSIS FOR

M.PHARM DISSERTATION

SUBMITTED TO

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

KARNATAKA.

BY

VUMMADI DILIP REDDY

DEPARTMENT OF PHARMACOLOGY,

NARGUND COLLEGE OF PHARMACY,

BANGALORE- 560085.

KARNATAKA.

(2010-2012)

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

KARNATAKA.

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR P.G. DISSERTATION

1. / Name of the candidate and address
(in block letters) / VUMMADI DILIP REDDY,
NARGUND COLLEGE OF PHARMACY,
DATTATREYA NAGAR, II MAIN,
100FT RING ROAD, BSK III STAGE,
BANGALORE-560085.
2. /

Name of the Institution

/ NARGUND COLLEGE OF PHARMACY,
DATTATREYA NAGAR, II MAIN,
100FT RING ROAD, BSK III STAGE,
BANGALORE-560058.
3. /

Course of study and subject

/

MASTER OF PHARMACY IN PHARMACOLOGY

4. /

Date of the admission

/ 9th March 2010
5.
6.
7.
8. /

Title of the topic:

EVALUATION OF WHEATGRASS JUICE FOR
ANTIDIABETIC ACTIVITY IN RATS
BRIEF RESUME OF THE INTENDED WORK
6.1. NEED FOR THE STUDY:
Diabetes mellitus, long considered a disease of minor significance to world health, is now taking its place as one of the main threats to human health in the 21st century.1 The past two decades have seen an explosive increase in the number of people diagnosed with diabetes worldwide.2,3 Estimates suggest that the world prevalence of diabetes among adults (aged 20–79 years) will be 6.4%, affecting 285 million adults, in 2010, and will increase to 7.7%, and 439 million adults by 2030. Between 2010 and 2030, there will be a 69% increase in numbers of adults with diabetes in developing countries and a 20% increase in developed countries.4
Diabetes mellitus is a group of metabolic diseases characterized by hyperglycemia resulting from defects in insulin secretion, insulin action, or both. The chronic hyperglycemia of diabetes is associated with long-term damage, dysfunction, and failure of various organs, especially the eyes, kidneys, nerves, heart, and blood vessels.5
In the management of diabetes mellitus, several approaches are often employed which include dietary intervention, use of different classes of oral hypoglycaemic agents, insulin injections, aerobic exercise and food supplements.6
Although, oral hypoglycemic agents/ insulin are the mainstay of treatment of diabetes and are effective in controlling hyperglycemia, they have prominent side effects and fail to significantly alter the course of diabetic complications. The common side effects associated with the main classes of drugs used for the treatment of type 2 diabetes mellitus are hypoglycemia, weight gain, gastrointestinal disorders, peripheral edema and liver disease.7
In view of these shortcomings, herbal pharmacotherapy is often explored by diabetic patients. Many natural products and herbal medicines have been recommended for the treatment of diabetes.8 In spite of the worldwide use of herbs and medicinal plants, the effective treatment of diabetes with phytochemicals has not been validated with scientific criteria which may support their substitution for the current therapy.9 Metformin is the only ethical drug so far approved for treatment of Type 2 diabetes and is derived from a medicinal plant (Galegos officinalis) historically used to treat diabetes.10 World Health Organization expert committee on diabetes has listed as one of its recommendations that traditional methods of treatment for diabetes should be further investigated.11
Hence in this context there is an immense need for search for novel herbal drugs which possess antidiabetic activity. So the present study was taken up to screen the antidiabetic potential of wheatgrass juice, an easily available and consumable plant.
6.2. REVIEW OF LITERATURE:
Wheatgrass is the young grass of common wheat plant, Triticum aestivum Linn., belonging to the family Poaceae (Gramineae). It is commonly known as “green blood” due to its high chlorophyll content which accounts for 70% of its chemical constituents.12
Wheatgrass juice is a rich source of vitamins A, C, E and B complex. It contains minerals like calcium, phosphorous, zinc, boron, and molybdenum. Various enzymes responsible for its pharmacological actions are protease, amylase, lipase, cytochrome oxidase, transhydrogenase, super oxide dismutase. The major clinical utility of wheatgrass juice is due to its antioxidant action which is derived from its high content of bioflavonoids like apigenin, quercetin, and luteolin. Other compounds present, which make this grass therapeutically effective, are the indole compounds, choline and laetrile.12
Orally administered fresh juice of Triticum aestivum leaves showed appreciable hypolipidemic properties in normal rats.13
In a study it was found that drinking wheatgrass juice helped produce healthier blood levels in cancer patients who are on chemotherapy, thus decreasing the need for blood building medications.14
The use of wheatgrass juice for treatment in active distal ulcerative colitis showed significant reduction in the overall disease activity and in the severity of rectal bleeding.15
In another in vitro study it was found that wheat sprout extract inhibited the metabolic activation of carcinogens and decreased their cancer causing ability by up to 99 percent.16
Wheatgrass juice has the potential to lower blood transfusion requirements in β-thalassemia major. A beneficial effect of wheatgrass was defined as decrease in the requirement of packed red cell by 25% or more.17 Wheatgrass juice is also useful in adjuvant therapy in hemolytic anemia.12
Aqueous extracts of wheatgrass are good sources of antioxidants. Significant antioxidant activity was demonstrated by in vitro studies.18
Inclusion of wheatgrass in various food recipes resulted in a significant decrease in glycemic index in human subjects.19
6.3. OBJECTIVES OF THE STUDY:
The objective of this study is to evaluate the antidiabetic potential of fresh wheatgrass (Triticum aestivum) juice in rats.
MATERIALS AND METHODS
7.1. SOURCE OF DATA:
Data will be obtained from the experimental work, which includes laboratory based animal studies and evaluation of various parameters.
1.  Library: Nargund college of pharmacy library & RGUHS digital Library
2.  Internet source
3.  International and National Science and medicine abstracts
4.  International and National Science and medicine journals
7.2. METHODS OF COLLECTION OF DATA:
The data collected is based on animal experimentation as per the parameters studied under each animal model.
METHODOLOGY:
7.2.1. Streptozotocin induced diabetic animal models:20
·  Animals: Albino Wistar Rats.
·  Weight: 180-200gm.
·  Test drug: Wheatgrass juice.
·  Groups: 6 groups of 10 animals each.
§  Group I: Normal control - animals will be administered with vehicle orally
§  Group II: Diabetes induced control - animals will be administered with vehicle orally
§  Group III: Diabetes induced + Standard drug (Glibenclamide – administered orally at a dose of 600 µg/kg body weight)20
§  Group IV: Diabetes induced + Test drug treated dose I (5 ml/kg body weight, administered orally once a day)13
§  Group V: Diabetes induced + Test drug treated dose II (10 ml/kg body weight, administered orally once a day)13
§  Group VI: Test drug treated normal group (10 ml/kg body weight, administered orally once a day)
·  Procedure: The animals of group II, III, IV & V will be injected with streptozotocin (70 mg/kg, i.p.). Five days after injection, the rats with fasting blood glucose higher than 180 mg/dl will be used for the experiments.
·  Parameters to be evaluated:
§  Body weight.
§  Fasting blood glucose levels.
§  Histopathological findings of pancreas.
§  Lipid profile.
7.2.2.  Dexamethasone induced insulin resistant animal models:21
·  Animals: Albino Wistar Rats.
·  Weight: 180-200gm.
·  Test drug: Wheatgrass juice.
·  Groups: 6 groups of 10 animals each.
§  Group I: Normal control - animals will be administered with vehicle orally
§  Group II: Diabetes induced control - animals will be administered with vehicle orally
§  Group III: Diabetes induced + Standard drug (Glibenclamide – administered orally at a dose of 600 µg/kg body weight)20
§  Group IV: Diabetes induced + Test drug treated dose I (5 ml/kg body weight, administered orally once a day)13
§  Group V: Diabetes induced + Test drug treated dose II (10 ml/kg body weight, administered orally once a day)13
§  Group VI: Test drug treated normal group (10 ml/kg body weight, administered orally once a day)
·  Procedure: Dexamethasone sodium phosphate, at a dose of 10 mg/kg will be administered subcutaneously to the overnight-fasted rats of group II, III, IV & V once a day and continued till the end of experiment along with the standard and test drugs.
·  Parameters to be evaluated:
§  Body weight.
§  Fasting blood glucose levels.
§  Histopathological findings of pancreas.
§  Lipid profile.
STATISTICAL ANALYSIS:
The data will be analyzed by using one-way analysis of variance (ANOVA).
7.3. Does the study require any investigation or intervention to be conducted on patients or other human or animals?
Yes. The above study requires investigation on animals. The effect of drug will be studied on various parameters using rat as animal model.
7.4. Has ethical clearance been obtained from your institute?
Applied for the IAEC of Nargund college of pharmacy, Bangalore.
REFERENCES:
1.  Zimmet P, Alberti KGMM, Jonathan S. Global and societal implications of the diabetes epidemic. Nature2001;414:782-7.
2.  Amos AF, McCarty DJ, Zimmet P. The rising global burden of diabetes and its complications: estimates and projections to the year 2010. Diabet Med 1997;14:S1–S85.
3.  King H, Aubert RE, Herman WH. Global burden of diabetes, 1995-2025. Prevalence, numerical estimates, and projections. Diabetes Care 1998;21(9):1414–31.
4.  Shaw JE, Sicree RA, Zimmet PZ. Global estimates of the prevalence of diabetes for 2010 and 2030. Diabetes Res Clin Pract 2010;87(1):4-14.
5.  American diabetes association. Diagnosis and classification of diabetes mellitus. Diabetes Care 2005;28(1):S37-S42.
6.  Adeneye AA, Adeyemi OO. Further evaluation of antihyperglycaemic activity of Hunteria umbellate (K. Schum) Hallier f. seed extract in experimental diabetes. J Ethnopharmacol 2009;12(2):238–43.
7.  Geetanjali K, Santosh S, Rakesh KK, Naik SN. Commonly consumed Indian plant food materials in the management of diabetes mellitus. Diabetes & Metabolic Syndrome: Clinical Research & Reviews 2010;4(1):21–40.
8.  Jung M, Park M, Lee HC, Kang YH, Kang ES, Kim SK. Antidiabetic agents from medicinal plants. Current Medicinal Chemistry 2006;13:1203–18.
9.  Frode TS, Medeiros YS. Animal models to test drugs with potential antidiabetic activity. J Ethnopharmacol 2008;115:173–83.
10.  Luo J, Fort DM, Carlson TJ, Noamesi BK, nii-Amon-Kotei D, King SR, et al. Cryptolepis sanguinolenta: an ethnobotanical approach to drug discovery and the isolation of a potentially useful new antihyperglycaemic agent. Diabet Med 1998;15:367–74.
11.  World Health Organization, Expert committee on diabetes mellitus, second report. Technical report series 646.WHO, Geneva, 1980.
12.  Swati P, Sushma D, Indira R, Alka G, Mamta D. Multitude potential of wheatgrass juice (Green Blood): An overview. Chronicles of young scientists 2010;1(2):23-8.
13.  Kothari S, Jain AK, Mehta SC, Tonpay SD. Effect of freshTriticum aestivumgrass juice on lipid profile of normal rats. Indian J Pharmacol 2008;40(5):235-6.
14.  Bar-Sela G, Tsalic M, Fried G, Goldberg H.Wheatgrass juice may improve hematological toxicity related to chemotherapy in breast cancer patients: A pilot study.Nutr Cancer2007;58(1):43-8.
15.  Ben-Arye E, Goldin E, Wengrower D, Stamper A, Kohn R, Berry E.Wheatgrass juice in the treatment of active distal ulcerative colitis: A randomized double-blind placebo-controlled trial.Scand J Gastroenterol 2002;37(4):444-9.
16.  Lai CN, Dabney B, Shaw C. Inhibition of in vitro metabolic activation of carcinogens by wheat sprout extracts. Nutr Cancer 1978;1(1):27-30.
17.  Marwaha RK, Bansal D, Kaur S, Trehan A.Wheatgrass juice reduces transfusion requirement in patients with thalassemia major: A pilot study.Indian Pediatr 2004;41(7):716-20.
18.  Kulkarni SD, Tilak JC, Acharya R, Rajurkar NS, Devasagayam TP, Reddy AV. Evaluation of the antioxidant activity of wheatgrass (Triticum aestivum L.) as a function of growth under different conditions. Phytother Res 2006;20(3):218-27.
19.  Uma I, Minal S, Swati D, Uliyar VM. Glycemic and lipemic response of wheat grass incorporated recipes. Journal of Herbal Medicine and Toxicology 2010;4(1):161-4.
20.  Eidia A, Eidib M, Esmaeilia E. Antidiabetic effect of garlic (Allium sativum L.) in normal and streptozotocin-induced diabetic rats. Phytomedicine 2006;13:624–9.
21.  Shalam MD, Harish MS, Farhana SA. Prevention of dexamethasone- and fructose-induced insulin resistance in rats by SH-01D, a herbal preparation. Indian J Pharmacol 2006;38(6):419–22.
9. / SIGNATURE OF THE CANDIDATE /
VUMMADI DILIP REDDY
10. / REMARKS OF THE GUIDE / Recommended for registration
11. / NAME AND DESIGNATION OF
11.1. GUIDE / PROF. M.S.HARISH
PROFESSOR
DEPARTMENT OF PHARMACOLOGY
NARGUND COLLEGE OF PHARMACY
11.2. SIGNATURE
11.3. CO-GUIDE / ------
11.4. SIGNATURE / ------
11.5. HEAD OF THE DEPT. /
Dr. H.J.HRISHIKESHAVAN
PROFESSOR & HEAD OF THE DEPARTMENT
DEPARTMENT OF PHARMACOLOGY
NARGUND COLLEGE OF PHARMACY
11.6. SIGNATURE
12. / REMARKS OF THE PRINCIPAL /
Recommended for registration
12.1. SIGNATURE