“EVALUATION OF ANTIDIABETIC ACTIVITY OF HELICTERES ISORA LINN FRUITS ON ALLOXAN INDUCED DIABETIC MICE”
Synopsis for registration of M.Pharm Dissertation submitted to
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BENGALURU
KARNATAKA
Submitted by
A RAJENDRA PRASAD
1st Year M.Pharm.
Under the Guidance of
Dr. SHANMUGA SUNDARAM M.Pharm, Ph.D
HOD, Dept of Pharmacology
ADITYA BANGLORE INSTITUTEOF PHARMACY EDUCATION&RESEARCH,
BENGALURU-64
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
KARNATAKA, BENGALURU.
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. / Name of the candidate and address / A RAJENDRA PRASADM.PHARM, PART-I,
DEPARTMENT OF PHARMACOLOGY,
ADITYA BENGALURU INSTITUTE FOR PHARMACY EDUCATION & RESEARCH, 12, KOGILU MAIN ROAD, YELAHANKA.
2. / Name of the Institution / ADITYA BANGALORE INSTITUTE FOR PHARMACY EDUCATION & RESEARCH, BENGALURU.
3. /
Course of study and subject
/ M.Pharm-Pharmacology4. /
Date of the admission
/ June-20115. /
Title of the topic
“EVALUATION OF ANTIDIABETIC ACTIVITY OF HELICTERES ISORA LINN FRUITS ON ALLOXAN INDUCED DIABETIC MICE”6.0 /
Brief resume of the intended work
7.08.0 / 6.1 - Need for the study:
Diabetes mellitus is one of the common metabolic disorders and 1.5% of the total population suffers from this disease throughout the world. Insulin and oral hypoglycemic agents like sulphonylureas and biguanides are still the major players in the management of the disease. However complete cure of the disease has been eluding physicians for centuries and the quest for the development of more effective anti-diabetic agent is pursued relentlessly1.
Many herbal products have been described for the use of diabetes mellitus in ancient literature; herbal preparations alone or in combination with oral hypoglycemic agent sometimes produce a good therapeutic response in some resistant cases wherein the allopathic drug alone has failed to produce the satisfactory results.
Helicteres isora Linn which belongs to family Sterculiacae is a sub-deciduous shrub or small tree of spreading habit with stem 1-5inches diameter, reaching a height of 5-15 feet. The species is native to Asia and Australia2.It occurs throughout India, from Jamuna eastwards to Nepal, Bihar, Bengal, southern India and Andaman islands.
The fruits are astringent, acrid, refrigerant, demulcent, constipating, stomachic, vermifuge, vulnerary, haemostatic and urinary astringent, they are useful in vitiated conditions of pitta, colic, flatulence, diarrhoea, dysentery, Verminosis, wounds, ulcers, hemorrhages, epistaxis and diabetes3. The root and stem barks are considered to be expectorant demulcent, astringent and anti-galactagogue and are useful in colic, scabies, empyema, gastropathy, diabetes, diarrhea and dysentery3,4. Tradionally, the root juice is claimed to be useful in diabetes, empyema, and a favorite cure for snakebite4,5.
The fruits were studied phytochemically and the presence of tannins, cardiac glycosides, sterols, triterpenes, α and β amysin, lupeol, friedelin, taraxerone and β-sitosterol have been demonstrated6.
Though the fruits are claimed to possess antidiabetic activity, their systematic investigation
has not been carried out so far. Hence it was thought worthwhile to investigate the antidiabetic activity of these fruits and to validate the claims made in the Indian traditional systems of medicine.
6.2 - Review of Literature:
Alloxan-induced diabetes consistently produced the main characteristics of diabetes mellitus including polydipsia, polyphagia, polyuria, decreased insulin levels, weight loss and hyperglycemia. This study evaluated the blood glucose levels, urinary glucose, body weight, feed intake and water intake in experimental diabetes induced by alloxan in mice7.
The fruits have shown a stimulant effect on rat uterus and a slight spasmogenic activity on isolated guinea-pig ileum8,the fruit extract possess weak anti-HIV (I) activity9, the aqueous extract of the bark showed significant hypoglycaemic effect10 ,lowering effect of hepatic enzymes11, glycoprotein levels12, antihyperglycaemic and antiperoxidative13, reinstate brain antioxidant enzymes14, antioxidant activity in diabetic rats15, hypolipidaemic effect16and roots show antinociceptive activity17,antidiabetic and hypolipidemic activity18.Cucurbitacin B and isocucurbitacin B were isolated and reported to possess cytotoxic activity19.
6.3 – Objective of Study:
The aim of the study is to evaluate the antidiabetic effect of the crude fruit extract preparation (Helicteres isora linn) in experimental type-2 diabetes on alloxan induced diabetic mice.
The objectives of the research are:
1. Acute toxicity studies
2. To estimate the blood sugar in control as well as treated animals (mice).
3. To estimate lipid profile in mice.
4. To study the histopathology of pancreas in the animals.
5. To study the serum biomarkers levels like ALT, AST, LDH and CK-MB.
MATERIALS AND METHODS
PLANT MATERIAL:
The fruits will be collected from Nilgiris, Ooty, Tamil Nadu and authenticated.
EXTRACTION :
The fruits of the plant would be dried in shade and extracted with 70% aqueous alcohol by kinetic maceration. The extracts of the plant would be filtered and collected, and concentrated by using a Rotatory Flash Evaporator.
7.1 – Source of Data:
Whole work is planned to generate data from laboratory studies i.e. experiments are performed as described in references. Experimental studies in journals and in text books available with college.
Web sites: www.sciencedirect.com
www.pubmed.com
www.google.com
www.ijp-online.com
7.2 – Method of Data Collection:
The Experiments will be conducted using different animal models and the data will be generated from such experimental studies.
METHODOLOGY:
Experimental animals:
In-house laboratory bred healthy male albino mice of Wistar strain weighing 25-40 g will be included for the study. Animals will be housed in polypropylene cages on clean paddy
husk bedding. Animals will be maintained under controlled temperature at 250C ± 20C with 12 hr light/dark cycle. All animals will have a free access to food and water ad libitum.
Administration of Drug:
The extract will be suspended in 1% w/v CMC and the suspension will be orally administered to the animals once daily for seven days. Three doses will be selected depending on the toxicity profile of the extract upon the oral treatment.
Induction of diabetes:
Alloxan monohydrate was injected intraperitoneally at dose of 200 mg/kg body weight (BW).The induction of alloxan-induced diabetes was confirmed by measuring the blood glucose level. Mice with glucose levels above 200 mg/dl were subjected to further evaluation with a glucose challenge and a standardized oral glucose tolerance test to confirm diabetes mellitus.
Parameters estimated:
The serum separated from the blood will be used to estimate the blood glucose and lipid profile. The biomarker levels viz., Alanine transaminase (ALT), Aspartate transaminase (AST), Lactate dehydrogenase (LDH) and Creatinine kinase-MB (CK-MB) will also be measured using the standard procedures20.
Other Parameters:
· Body weight.
· Histopathology of pancreas.
Method of collection of blood samples:
Blood samples of about 2ml will be collected from retro-orbital sinus under light ether anaesthesia and the serum will be separated from blood by centrifuging at 6000rpm for 15min.
number of groups to be used in the study
Group 1: Control (saline-0.5 ml/kg)
Group 2: Herbal extract in normal animals
Group 3: Type-2 diabetic group (Alloxan induced mice)
Group 4: Diabetic + herbal extract (First dose)
Group 5: Diabetic + herbal extract (Second dose)
Group 6: Diabetic + Standard allopathic drug
Number Of Animals Required:
No. of groups 06
No. of animal in each group 10
Total 10 X 6 = 60 animal
Statistical Analysis:
The statistical significance of the results will be analyzed by student‘t’ test and ANOVA. p<0.05 will indicate the significance of the result.
7.3 - Does The Study Require Any Investigation To Be Conducted On Patients Or Animals? If So, Please Describe Briefly.
Yes, the entire experimental models require usage of laboratory animals.
7.4 - Has Ethical Clearance Been Obtained From Your Institution In Case of 7.3?
Applied for IAEC approval. The same will be submitted once get the approval.
LIST OF REFERENCES:-
1. Gosh R, Sharatchandra KH, Rita S, Thokchom S. Hypoglycemic activity of Ficus hispidia (bark) in normal and diabetic albino rats. Ind J. pharmacol, 2004; 38(4):222-5.
2. Bhattacharjee S.K., Handbook of medicinal Plants, Pointer publishers, Jaipur, 1998, 179-80.
3. Warrier P.K., and Nambiar V.P.K., Indian Medicinal Plants, Orient Longman Ltd., 1995;3:132-5.
4. Kirithikar K.R., Basu, B.D., An, I.C.S., 1995. Indian Medicinal Plants. Vol.1, International Book Distributors, Dehradun, India, P. 371 – 2.
5. Singh, S.B., Singh, A.K., Thakur, R.S. Chemical constituents of the leaves of Helicteres isora. Ind J. Pharm Sci, 1984;46:148–9.
6. Deshmukh VK, Pandit U. Study of fruits of Helicteres. isora Linn. Ind J Pharmacol, 1968;30(12):283
7. Tanquilut N C, Tanguilut M R C, Estacio M.A.C, E.B.Torres, Rosario J.C. and Reyes B.A.S. Hypoglysemic effect of Lagerstroemia speciosa(L.) Pers. On alloxan-induced diabetic mice. J of Med Plants Research 2009;3(12):1066-71.
8. Pohocha N. and Grampurohit N.D. Antispasmodic activity of the fruits of Helicteres isora Linn .Phytother Res, 2001;15(1):134-9.
9. Otake T., Mori H., Morimoto M., Ueba N., Sutardjo S., Kusumoto I.T., and Hattori M., Phytother Res, 1995;9(1):6-10.
10. Kumar, G., Sharmila Banu, G., Murugesan, A.G., Rajasekara Pandian, M., Hypoglycaemic effect of Helicteres isora bark extracts in rats. J. Ethnopharmacol. 2006;107:304-7.
11. Kumar, G., Murugesan, A.G., Rajasekara Pandian, M., Effect of Helicteres isora bark extract on blood glucose and hepatic enzymes in experimental diabetes. Pharmazie,2006;61:353–5.
12. Kumar, G., Murugesan, A.G., Influence of Helicteres isora bark extracts on plasma and tissue glycoprotein compounds in streptozotocin diabetic rats. J. Clin. Diagn. Res. 2007;4:330-8.
13. Kumar, G, Sharmila Banu, G, Murugesan, A.G., Rajasekara Pandian, M., Antihyperglycaemic and antiperoxidative effect of Helicteres isora L.bark extracts in streptozotocin-induced diabetic rats. J. Appl. Biomed. 2007;5:97–104.
14. Kumar, G., Sharmila Banu, G., Murugesan, A.G., Rajasekara Pandian, M., Effect of Helicteres isora bark on brain antioxidant status and lipid peroxidation Streptozotocin diabetic rats.Pharma Biol, 2007e; 45:753-759
15. Kumar, G., Sharmila Banu, G., Murugesan, A.G., Rajasekara Pandian, M., Effect of Helicteres isora bark extract on streptozotocin induced diabetic rats. J.Trop. Med.Pl. 2007;8:27-33.
16. Kumar, G., Murugesan, A.G., Hypolipidaemic activity of Helicteres isora bark extracts in streptozotocin induced diabetic rats J. Ethnopharmacol,2008;116:161-6.
17. SamaVenkatesh, K.SaiLaxmi, B.Madhava Reddy, M.Ramesh.Antinociceptive activity of Helicteres isora. Fitoterapia 2007; 78:146–8.
18. Bean, M.F., Antoun, M., Abramson, D., Chang, C.J., Mc Laughlin, J.L., Cassady, J.M.,.Cucurbitacin B and Isocucurbitacin B Cytotoxic components of Helicteres isora. J. Nat.Prod. 1985; 48(3): 500-3.
19. Ranjan Chakrabarti, Reeba K. Vikramadithyan, Ramesh, Mullangi, V.M. Sharma, H. Jagadheshan, Y.N. Rao, P. Sairam, R. Rajagopalan. Antidiabetic and hypolipidemic activity of Helicteres isora in animal models. J. Ethnopharmacol. 2002;81:343-349
20. Raghavan B, Kumari SK. Effect of Terminalia Arjuna stem bark on antioxidant status in liver and kidney of alloxan diabetic rats. Ind J. physiol. Pharmacol. 2006; 50(2):133-42.
9 / Signature of the candidate /
(A RAJENDRA PRASAD)
10 / Remarks of the Guide: / Recommended for research
11 / Name and Designation of:
11.1 Guide:
11.2 Signature: / Dr. SHANMUGA SUNDARAM M.Pharm, Ph.D
HOD, Dept of Pharmacology
11.3 Co-Guide:
11.4 Signature / Not applicable
11.5 Head of the Department:
11.6 Signature / Dr. SHANMUGA SUNDARAM M.Pharm, Ph.D
HOD, Dept of Pharmacology
12 / 12.1 Remarks of the Chairman and Principal / Recommended for research
12.2 Signature / Dr. VISHWANATH .M.Pharm, Ph.D
PRINCIPAL
ADITYA BENGALURU INSTITUTE FOR PHARMACY EDUCATION & RESEARCH BENGALURU-64