<p> RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE, KARNATAKA,</p><p>ANNEXURE II PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION</p><p>1. Name of the Candidate Suma K.G. and Address a. Postal Address Al-Ameen College of Pharmacy, Hosur road, Opp to Lalbagh main gate, Bangalore-560027 Karnataka, India.</p><p> b. Permanent Address D/O k.Gangadharappa, Retired VA, 2ND Main 2ND Stage, Krushinagar, Shimoga-577201, Karnataka, India.</p><p>2. Name of the Institute Al-Ameen College of Pharmacy, Hosur road, Opp to Lalbagh main gate, Bangalore-560027 Karnataka,India.</p><p>3. Course of Study and Subject M.Pharmacy in Pharmacology.</p><p>4. Date of Admission to Course 01-08-2011</p><p>5. Title of the Topic: </p><p>“Pharmacological evaluation of the effect of phloroglucinol in rat model of diabetic nephropathy”. </p><p>1 6. Brief resume of the intended work: 1 Need for the study: Diabetic nephropathy is partial loss of function of kidney associated with nephrotic syndrome, glomerulosclerosis, persistent albuminuria and declining glomerular filtration rate.1</p><p>Diabetic nephropathy is major cause of chronic kidney disease. In India about 30% of diabetic patients are prone to chronic kidney disease.2 </p><p>Hence strategies to control and prevent diabetic nephropathy will result in decreased patient morbidity and mortality.</p><p>6.2 Review of literature</p><p>Diabetic nephropathy can occur in both type1 and type2 diabetes mellitus. The peak onset of nephropathy is between is between 10-15 years after the onset of disease. </p><p>Multiple biochemical mechanisms have emerged to explain adverse effect of hyperglycemia including protein kinase C(PKC),mitogen activated protein kinase(MAPK),polyol pathway, advanced glycation end products and oxidative stress.3</p><p>Among these , oxidative stress has been considered as a potential mechanism for diabetic kidney disease because oxidative stress promotes the formation of AGE as well as PKC-mark activation.</p><p>Phloroglucinol (1,3,5 benzentriol) is a monomeric unit of phloratannins. Natural phloroglucinol is obtained from fresh fruits of Eucalyptus maiden belonging to family Myrtacae.</p><p>Phloroglucinol has shown several biological effects like antioxidant, anti-inflammtory,anti- thrombotic, spasmolytic,enzyme inhibitory and anticancer activity.Since phloroglucinol exhibits antioxidant activity and anti-inflammatory activity this study was undertaken to evaluate the effect of phloroglucinol on nephropathy in diabetic rats.</p><p>Ionizing radiation induced cell damage, can be protected by phloroglucinol through inhibition of oxidative stress invitro and invivo. Hence phloroglucinol shows protective action through antioxidant activity.5</p><p>Protective effect of phloroglucinol against myocardial ischaemia- reperfusion injury is related to inhibition of myeloperoxidase activity and inflammatory cell infiltration.6</p><p>Phloroglucinol has antithrombotic and profibrinolytic activities.7 </p><p>2 Phloroglucinol has been used to delivery to shorten the duration of labour and lighten the degree of pain in humans. This proves the sposmolytic activity of phloroglucinol.8</p><p>Through enzyme inhibitory activity phloroglucinol can protect the gastric mucosa against ethanol induced –injury which is related to inhibiting the MPO activity and increasing the catalase activity in gastric tissue.9</p><p>6.3 Objective of study:</p><p>The aim of this study is to evaluate the effect of phloroglucinol in rat model of diabetic nephropathy.</p><p>The objectives of research are:</p><p>1. To study the nephroprotective effect of phloroglucinol by measuring serum creatinine, serum </p><p> urea, blood urea nitrogen and histopathology of kidney in diabetic rat.</p><p>2. To study the curative effect of phloroglucinol by measuring serum creatinine, serum urea, </p><p> blood urea nitrogen and histopathology of kidney in diabetic rat.</p><p>3. To evaluate the invivo antioxidant effect of phloroglucinol in kidney homogenates of diabetic </p><p> rats. </p><p>4. To evaluate the effect of phloroglucinol on hyperglycaemia and body weight in diabetic rats. 7. MATERIALS AND METHODS: 7.1 Source of Data: Information centre Al-Ameen College of Pharmacy.</p><p>Web Sites: www.sciencedirect.com www.pubmed.com www.ijp-online.com www.elsevier.com www.google.com</p><p>3 Journals: J Am Soc Nephrol Clinical and Experimential Pharmacology and Physiology Kidney International Fundamental and Clinical Pharmacology British Journal of Nutrition Journals of Biomedical Nanotechnology Indian Journal of Pharmacology JPP JPET</p><p>7.2 Method of Collection of Data: The data collected will be based on animal experimentation as per the parameters studied under each animal model, which are mentioned under the objectives of the study.</p><p>EXPERIMENTAL MODELS</p><p>Experimental animals and diet:</p><p>Adult albino rats of Wistar strain weighing 160-200g will be included for the study. Animals will be housed in polypropylene cages on clean paddy husk bedding. Animals will be kept in an environment with controlled temperature at 250C±20C and exposed to 12hr light/dark cycle. All animals will be provided with fresh food and water ad libtium.</p><p>Induction of diabetes10: </p><p>Overnight Rats will be injected with Nicotinamide 120 mg/kg by i.p route and after 15 min STZ 60mg/kg by i.p will be injected.Blood glucose will be estimated 72hrs post STZ injection and rats having >250 mg/dL11 blood glucose level will be included in the study. </p><p>Grouping:</p><p>4 Preventive study: Diabetic rats will be administered with phloroglucinol for 35 days , post induction of diabetes as a treatment. Group 1: Negative control no treatment. Group 2: Positive control, diabetic rats administered vehicle per oral. Group 3: Diabetic rats administered phloroglucinol 10mg/kg per oral daily for 35 days.12 Group 4: Diabetic rats administered phloroglucinol 30mg/kg per oral daily for 35 days.12 Group 5: Diabetic rats administered vitamin E(standard reference) 250mg/kg per oral daily for 35 days.13,14 Curative study: Rats after induction of diabetes will be left for 35 days for development of diabetic nephropathy. Post treatment which will be given for 30days.</p><p>Group 1: Negative control no treatment. Group 2: Positive control,diabetic rats administered vehicle per oral. Group 3: Diabetic rats administered phloroglucinol 10mg/kg per oral from day 35 to day 65. Group 4: Diabetic rats administered phloroglucinol 30mg/kg per oral from day 35 to day 65. Group 5: Diabetic rats administered vitamin E(standard reference) 250mg/kg per oral daily from day 35 to day65.</p><p>7.3Parameters measured 1. Serum creatinine15, serum urea15, Blood urea nitrogen15, serum glucose16. 2. Antioxidant enzymes in kidney homogenization, superoxide dismutase (SOD),Catalase(CAT),Glutathione(GSH) 17. 3. Histopathology of kidney15. 4. Body weight and kidney weight18. </p><p>7.4 Does the study require any investigation or interventions to be conducted on patients or the human or animals? If so please describe briefly: Yes, Study requires investigation on animals. The effects of the drug will be studied on various parameters using rats as experimental animal model.</p><p>7.5 Has ethical clearance been obtained from your institute Yes it has been approved. 8. List of References: 1. Pitchal B, Vishal AC, Vijay K, Akash J, Jayarami R, Manjeet S.Experimental models for nephropathy. Journal of Rennin-angiotensin-aldosterone system.2008; 40(9):189-</p><p>5 194. </p><p>2. Anubav T, Manish M, Vaibhav S, Hemant K, Amit NDD, Ram CS, Kamlakar T.Determinants of Intravascular Resistance in Indian Diabetic Nephropathy Patients: A Hospital-Based Study. International Journal of Vascular Medicine.2011;1- 7. </p><p>3. Daisuke K, Kazuyuki H, Munehiro K, Atsunori K, Ryuichi K, Masakazu H,Effect of antioxidants in diabetes-induced oxidative stress in the glomeruli of diabetic rats.J Am Soc Nephrol. 2003; 58(14):250-253.</p><p>4. Inder PS, Jasmeen S, Parikshit B and William JF. Phloroglucinol compounds of therapeutic interest: Global patent and technology status. Informa health care. 2009;19(6):847-866</p><p>5. Kyoung AK, Rui Z, Sungwook C, Su JL, Jihoon K, Jeongtae K,et al., Phloroglucinol(1,3,5-trihydroxy benzene) protects against ionising radiation-induced cell damage through inhibition of oxidative stress invitro and invivo.Chem Bio Interact. 2010;185(3):215-226.</p><p>6. Ting TL,Yi SZ,Lan H,Nian SL, Jun P, Yuan JL. Protective effect of Phloroglucinol against myocardial ischaemia- reperfusion injury is related to inhibition of myeloperoxidase activity and inflammatory cell infiltration.Clinical and Experimental Pharmacology and Physiology. 2011;38(1):27-33.</p><p>7. Jong SB. Antithrombotic and Profibrinolytic activities of Phloroglucinol. Food and Chemical Toxicology. 2011;49(7):1572-1577.</p><p>8. Samina T, Bilqis A, Zahid A. Phloroglucinol for acceleration of labour: Double blind, randomized controlled trial. JPMA.2005;55(7):273-273.</p><p>9. Nian SL, Xiu JL, Yi SZ, Lan H, Yin ZL, Jun P. Phloroglucinol protects gastric mucosa against ethnol -induced injury. Fundamental and Clinical Pharmacology.2011;25(4):462-468.</p><p>10. Punitha ISR, Rajendran K, Arun S, Annie S. Alcoholic stem extract of coscinium fenestratum regulates carbohydrate metabolism and improves antioxidant status in streptozotocin-nicotinamide induced diabetic rats. Advanced access publications.2005;2(3):375-381.</p><p>11. Zhang XF, Tan BKH, Effects of an Ethanolic Extract of Gynura procumbens on Serum Glucose, Cholesterol and Triglyceride Levels in Normal and Streptozotocin-</p><p>6 Induced Diabetic Rats.SMJ.2000;41(1)</p><p>12. Ting TL, Yi SZ, Lan H,Nian SL, Jung P, Yuan JL.Protective effect of phloroglucinol against myocardial ischaemia-reperfusion injury is related to inhibition of myeloperoxidase activity and inflammatory cell infiltration.Clin Exp Pharmacol Physiol.2011:38(1):28-33.</p><p>13. Ashraf B, Abdel N, Mohamed H, Abdel W, Foad FA. Protective effects of vitamin E and probucol against gentamycin nephrotoxicity in rats. Pharmacol Res 1999;40(2):183-187.</p><p>14. Hyun SL, Jun YJ, Bong CK, Young SK, Yuan ZZ, Hong KC.Dietary antioxidant inhibits lipoprotein oxidation and renal injury in experimental focal segmental glomerulosclerosis. Kidney international 1997;51(3):1151-1159.</p><p>15. Matthew DB, Erwin B, Frank CB, Thomas MC, Raymond CH, Kumar S, et al. Mouse model of diabetic nephropathy.J Am Soc Nephrol 2005; 16:27-45.</p><p>16. Guo GW, Xiao HL, Wei.L, Xue Z, Cui Z.Protective effect of luteolin on diabetic nephropathy in STZ induced diabetic rats. Evidence based complementary and alternative medicine.2011:1-7.</p><p>17. Osama MAS, Rawhia HEE, Gamal EIH, Nermin EH, Mabrouk MG. Experimental diabetic nephropathy can be prevented by propolis:Effect on metabolic disturbences and renal oxidative parameters.Pak.J.Pharm.2009; 22(2);205-210.</p><p>18. Derakhshanfar A, Bidadkosh A, Kazeminia S. Vitamin E protection against gentamicin induced nephrotoxicity in rats: A biochemical and histopathologic study. Iranian journal of veterinary research 2007;8(3):231-238.</p><p>9. SIGNATURE OF THE CANDIDATE:</p><p>7 10. REMARKS OF THE GUIDE: “Pharmacological evaluation of the effect of phloroglucinol in rat model of diabetic nephropathy”. to be carried out by Suma.K.G of M.Pharm has been discussed and worked out under my directions and supervision as an official guide. The project work envisaged is of great importance in the field of pharmacology. The work can be carried out in pharmacology laboratory of Al-Ameen College of Pharmacy for which facilities are available. Hence the project is viable and is recommended for clearance and approval. </p><p>11 11.1 Name and Designation of Guide Dr. . Rema Razdan, Professor, Department of Pharmacology, Al-Ameen College of Pharmacy. Bangalore-560027. 11.2 Signature</p><p>Dr. Md. Naseeruddin Inamdar. 11.3 Head of the Department Head of the department, Department of pharmacology, Al-Ameen college of pharmacy, Bangalore-560027.</p><p>11.4 Signature</p><p>12. 12.1 Remarks of the Principal</p><p>Recommended for approval 12.2 Signature</p><p>Prof.B.G.Shivananda Principal, AL-Ameen College of Pharmacy,Bangalore-560027</p><p>8</p>