RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE KARNATAKA

ANNEXURE II

PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION

1. / NAME OF CANDIDATE
And Address (in block letters) / Dr. SUSHEELA SOMAPPA HALEMANI,
D/O S.G.HALEMANI,
A.P.M.C STAFF QUARTERS,
BANGALORE ROAD,
BELLARY- 583101.
KARNATAKA.
2. / NAME OF THE INSTITUTION / J.J.M.MEDICAL COLLEGE,
DAVANGERE-577004.
3. / COURSE OF STUDY & SUBJECT /

MEDICAL

M.D. PHARMACOLOGY.

4. / DATE OF ADMISSION TO COURSE / 30th MAY 2011
5. /

TITLE OF TOPIC

/ “EVALUATION OF THE HYPOGLYCEMIC ACTIVITY OF AEGLE MARMELOS IN
ALLOXAN INDUCED DIABETIC ALBINO RATS”
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12. / BRIEF RESUME OF THE INTENDED WORK:
6.1 Need for the study:
Diabetes mellitus (DM) comprises a group of common metabolic disorders that share the phenotype of hyperglycemia. Depending on the etiology of DM, factors contributing to hyperglycemia may include reduced insulin secretion, decreased glucose utilization and increased glucose production. DM is the leading cause of end stage renal disease, non-traumatic lower extremity amputation, adult blindness, etc. The worldwide prevalence of DM has risen dramatically over the past two decades. The International Diabetes Federation, projects that 438 million individuals will have diabetes by the year 2030. Nearly 1.7%-5.2% of deaths are attributed to diabetes worldwide.[1]
In the past two decades, diabetes has emerged as a global health problem. India alone harbours around 40 million diabetic people. The prevalence in rural area is 2.4% and in urban areas, it is 4-11.6%. By 2030 it is estimated that 79.4 million people will be suffering from diabetes in India.[2]
Despite the availability of many anti diabetic medicines in the market, diabetes and its related complications like atherosclerosis, neuropathy, nephropathy and retinopathy are continued to be the major medical problems. Oral hypoglycemic agents such as biguanides, sulphonylureas and thiozolidinediones and various insulin preparations are available for the treatment of diabetes. However, they have side effects associated with their uses like lipodystrophy, resistance, weight gain, diarrhoea etc. There is a growing interest in herbal remedies because of their effectiveness, minimal side effects and relatively low costs.[3]
Aegle marmelos (Rutaceae) commonly called, as ‘Bael’ in Hindi and Bilva in Kannada is indigenous to India. Leaves, fruits, stem and roots of Aegle marmelos have been in ethnomedicine for several medicinal properties: Astringents, antidiarrhoeal, antidysenteric, aphrodisiac and as an antidote to snake venom.[4]
In this study, an effort will be made to investigate the hypoglycemic effect of an aqueous seed extract of “Aegle marmelos” and its comparision with Glibenclemide using glucometer. This may prove beneficial to the development of novel therapeutic approaches in treatment of diabetes.
6.2. Review of Literature :
Diabetes mellitus is a common metabolic disorder in which both genetic and environmental factors contributes to its pathogenesis and involves insufficient insulin secretion, reduced responsiveness to endogenous or exogenous insulin, increased glucose production and abnormalities in fat /protein metabolism.[5]
In diabetes mellitus insulin preparations are the drug of choice for type-I DM, where as oral hypoglycemic agents are the first choice in type-II DM followed by insulin. Currently available anti diabetic drugs include; insulin, sulfonylurea, meglitinide, biguanide, glitazone, and sitagliptin, pramilintide, exenatide.[6]
Study done by M.C. Babu and Ramdasan Kuttan reflects that methanolic extract of Aegle marmelos could reduce blood sugar by 54% of the initial value. Superoxide dismutase and glutathione levels were found to be increased on 12th day.[7]
Article by N. Kamalakkannan, P. Stanely Mainzen Prince describes that the water extract of the fruits of Aegle marmelos reduces blood glucose levels in Streptozotocin induced diabetic Wistar rats. Oral administration of the water extracts (125 and 250 mg/ kg) twice a day for 4 weeks resulted in significant reductions in blood glucose levels.[8]
Study was performed to evaluate the hypoglycaemic and antioxidant effect of aqueous extract of Aegle marmelos leaves (AML) on diabetic rats. The plasma (GST) glutathione transferase levels were raised in diabetic rats when compared to controls and plasma glucose level decreased at the end of four weeks.[9]
Study done by Achyut Narayan Kesari, Rajesh Kuamr Guptha, Santhosh Kumar Singh, Sandhya Diwakar, Geetha Watal has shown that Aegle marmeols at a dose of 250 mg/kg was found to be the most effective dose and it decreases blood glucose level (BGL) by 35.1% in normal healthy rats after 6 h of administration. Treatment of diabetic rats with fasting blood sugar (FBS) >250 mg/dl for 14 days with a dose of 250 mg/kg reduces the FBS by 60.84%.[10]
Neuroprotective role of Aegle marmelos has been demonstrated in a study, which describes that pyridoxine treated in combination with insulin and Aegle marmelose has a role in the regulation of insulin synthesis and release, normalising diabetic related stress and anxiety through hippocampal serotonergic function.[11]
Varied classes of compounds Coumarins (Marmelosin, marmesin, imperatorin), alkaloids (Aeglin, aegelenine), Tannins (skimmianine), Carotenoids and seed oils and other miscellaneous compounds have been isolated from this plant used in ethnomedicine to exploit its medicinal properties including antidiabetic, antiulcer, antioxidant, antimalarial, anti-inflammatory, anticancer, radioprotective, antihyperlipidaemic, antifungal, antibacterial and antiviral activities.[12]
6.3. Objectives of study:
1) To evaluate the hypoglycemic activity of aqueous seed extract of Aegle marmelos in alloxan induced diabetes in albino rats.
2) To compare the hypoglycemic activity of aqueous seed extract of Aegle marmelos with a currently used oral hypoglycemic drug i.e. glibenclamide.
MATERIALS AND METHODS:
7.1 Source of data:
Albino rats of both sex of average weight 170-220g aged 3-4 months, which will be bred in central animal house of J.J.M. Medical College Davangere.
Chemical and drugs:
Alloxan monohydrate(150mg/kg body weight. intraperitoneally)
Glibenclamide ( 0.5mg/kg body weight orally )
Aqueous seed extract of Aegle marmelos.(250mg/kg body wt)
7.2 Method of collection of data (including sampling procedure, if any)
Albino rats will be divided into 5 groups, each group containing 6 rats.
Inclusion Criteria :
Ø  Animals weighing 170-220g.
Ø  Age 3-4 months.
Ø  Healthy with normal behaviour and activity.
Exclusion Criteria:
Ø  Animals weighing less than 170g and more than 220g.
Ø  Age < 3 months and >4 months.
Ø  Pregnant female rats.
Ø  Animals used for other studies recently.
Methods:
In this study, 30 albino rats with FBS in the range of 80-115 mg/dl will be selected. Three groups each containing six rats will be induced diabetes with Alloxan (150mg/kg body wt) by intraperitoneal route. After 7 days of induction, the rats that develop a stable hyperglycemia with fasting blood sugar > 250 mg/dl will be selected for the study.
Aqueous extract of Aegle marmelos and glibenclemide solutions will be prepared freshly in normal saline at the required concentrations and administered orally for 28 days.
In this study animals will be divided two groups;
A)  Non-diabetic groups:
1)  Group-1 (control): Normal saline 0.5 ml.
2)  Group-2 (Test): Aqueous seed extract of Aegle marmelos dissolved in normal saline (250mg/kg body wt)
B) Alloxan induced diabetic groups:
3)  Group-3 (control): Normal saline 0.5 ml.
4)  Group-4 (Test): Aqueous seed extract of Aegle marmelos dissolved in normal saline (250mg/kg body wt).
5)  Group-5 (Standard): Glibenclamide 0.5mg/kg body wt in normal saline.
Parameter observed:
Fasting blood sugar.
Estimation of blood glucose:
Fasting blood glucose levels from experimental animals will be measured on 0, 1, 3, 7, 14, 21 & 28th day by using glucometer.
Statistical analysis :
After collecting raw data, the values in all the groups will be analyzed by using one-way analysis of variance (ANOVA) statistical test, followed by post hoc test for inter group comparisons.
7.3. Does the study require any investigations or interventions to be conducted on patients or other humans or animals? If so, describe briefly?
YES
The study involves laboratory animals (albino rats) which will be induced diabetes by intra peritoneal administration of Alloxan monohydrate. It also requires the estimation of FBS by drawing blood from rat tail vein.
7.4  . Has ethical clearance been obtained from your institution in case of 7.3?
YES
LIST OF REFERENCES:
1) Alvin CP, Diabetes mellitus. In: Kasper, Braunwald, Fauci, Hauser, Longo, Jameson. Harrison’s principles of internal medicine, Vol 2; 17 th ed. New York: McGraw Hill; 2008. p. 2275-2276.
2) Park K. Park’s textbook of preventive and social medicine. 20th edn. Jabalpur: Banarasidas Bhanot; 2009. p. 341-343.
3) Kumar BD, Mitra A and Manjunatha M. In vitro and in vivo studies of antidiabetic Indian medicinal plants. Journal of Herbal Medicine and Toxicology 2009; 3 (2): 9-14.
4) Nandkarni A K Indian Materia Medica. vol. I, third ed. Popular Prakshan, Bombay. p. 45–49.
5) Powers AC and Alessio D. Endocrine Pancreas and Pharmacotherapy of Diabetes Mellitus and hypoglycaemia .in: Bruton L, Chabner B, Knollman B. Goodman and Gilman’s the pharmacological basis of therapeutics.12th ed. New York: McGraw Hill; 2011. p.1237.
6) Satoskar R S, Bhandarkar S D, Nirmala N Rege: Insulin, oral anti diabetic drugs and pharmacotherapy of Diabetis Mellitus; Pharmacology and pharmaco therapeutics;21st ed, Mumbai; Popular prakashan;2009:877,892.
7) Sabu MC and Kuttan R. Antidiabetic activity of Aegle marmelos and its relationship with its antioxidant properties. Indian J Physiol Pharmacol 2004; 48(1): p. 81-88.
8) Kamalakkannan N, Mainzen Prince PS. Hypoglycaemic effect of water extracts of Aegle marmelos fruits in streptozotocin diabetic rats. Journal of Ethnopharmacology 2003; 87 (2003): p. 207–210.
9) Upadya S, Kshama K, Shanbhag. A study of hypoglycemic and antioxidant activity of Aegle marmelos in alloxan induced diabetic rats. Indian J Physiol Pharmacol 2004; 48 (4): p. 476-480.
10) Kesari AN, Gupta RK, Sing SK. Hypoglycemic and antihyperglycemic activity of Aegle marmelos seed extract in normal and diabetic rats. Journal of Ethnopharmacology 2006; 107 (2006): P. 374–379.
11) Abraham PM, Kuruvilla KP, Mathew J, Malat A, Joy S, Paulose CS. Alterations in hippocampal serotonergic and INSR function in streptozotocin induced diabetic rats exposed to stress; neuroprotective role of pyridoxine and Aegle marmelose. Abraham et al. Journal of Biomedical Science 2010; P. 17:78.
12) Dhankhar S, Ruhil S, Balhara M, Seema Dhankhar 2 and Chhillar AK. Aegle marmelos (Linn.) Correa: A potential source of Phytomedicine. Journal of Medicinal Plants Research. 2011 may 4; Vol. 5(9). p. 1497-1507.
SIGNATURE OF THE CANDIDATE
REMARKS OF THE GIUDE / The present study of hypoglycemic activity of Aegle Marmelos extract was not taken up by any post graduate of Rajiv Gandhi University of Health Sciences in previous 5 years to the best of my knowledge.
NAME AND DESIGNATION
OF (In Block Letters)
11.1 Guide
11.2 Signature
11.3 Co-guide(If any)
11.4 Signature
11.5 Head of the Department
11.6 Signature. / DR. SURYANARAYANA BABUSHAW.N
M.D.,
PROFESSOR,
DEPARTMENT OF PHARMACOLOGY,
J.J.M. MEDICAL COLLEGE,
DAVANAGERE-577004.
DR. SOMASHEKAR. H.S
M.D
PROFESSOR AND H.O.D.,
DEPARTMENT OF PHARMACOLOGY,
J.J.M. MEDICAL COLLEGE,
DAVANAGERE-577004
12.1 REMARKS OF THECHAIRMANAND PRINCIPAL
12.2 Signature.

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