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DYSLIPOPROTEINEMIA AND OXIDATIVE STRESS IN ALLOXAN INDUCED DIABETIC RATS

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Supervisor Dr. R.K. Singh Prof & Head Department of Biochemistry Chattrapati Sahuji Maharaj Medical University (Formerly- King George’s Medical University) Lucknow- India

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“ Any species capable of independent existence that contains one or more unpaired electrons”. During metabolism molecular oxygen consumed by aerobic cells is reduced to water. Oxygen is partially reduced it becomes ‘activated’. This partial reduction leads to successive formation of reactive oxygen metabolites. Reactive Oxygen Species Introduction

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Diabetes mellitus, a common metabolic disorder, is one of the main cause of morbidity, mortality and third leading motive of death in many developed countries. Diabetic dyslipoproteinemia is main cause of heart attacks in diabetic patients. Diabetes is one of the major cause of blindness, renal failure, stroke and nephritis etc. Diabetes is as old as mankind and its incidence is considered to be high all over the world (Picup and Williams, 1997).

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Prevalence of diabetes in India is 2.4% in rural and 8.2% in urban areas. (Ramachandran et al . 1993) The rate of prevalence increases per year. It has been reported that in February, 2005 approximately 60 million of people in India are suffering from this disease (Yusuf, 2005). The most common lipid abnormalities in diabetes are hypertriglyceridemia and hyper cholesterolemia (WHO., 2006).

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type-I diabetes mainly occurs in childhood and puberty and is characterized by absolute insulin deficiency. Type-II diabetes usually develops in adults over age 40, and is characterized by insulin resistance (Berdanier, 2001).

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Moreover it has been suggested that the deficiency of antioxidant and trace elements such as Zn, Cu, Cr, Ni, Mn may also play a role in diabetes mellitus with dyslipoproteinemia (Feher et al ., 1982). Free oxygen radicals and oxidative stress is responsible for diabetic dyslipoproteinemia.

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Severe hyperglycemia cause generation of reactive oxygen species which are involved in peroxidative degradation of lipid. Retinal oxidative stress is increased in diabetic patients and rats due to down regulation of antioxidant defence enzymes like glutathione reductase (GSH) (Li et al ., 1999; Kowluru et al ., 1999). Prevention of diabetes still lies in realm of future on until then tens of million will continue to suffer from this disease.

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AIM AND OBJECTIVES The aim of present study, to develop drugs which are not only hypoglycemic but also antioxidant and lipid lowering property in diabetic rats. Natural products are the best option to treatment of diabetic dyslipoproteinemia. Accordingly we have selected four medicinal plants such as Anthocephalus indicus (Kadam), Hibiscusrosa sinensis (Gurhal), Cassia tora (Chakvat), Tinospora cordifolia (Gurch), which are known to posses potent biological activities. The effects of these plant materials will be compared to that of Gliblenclamide (A known hypoglycimic drug).

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In view of the above consideration, the present study has been planned to investigate the status of various biochemical parameters in alloxan induced diabetic rats, following treatment with various herbal hypolipidemic agents To investigate level of Blood glucose To assess the following lipid profile in serum lipoprotein (VLDL, LDL and HDL) Cholesterol Triglyceride Phospholipids Apolipoprotein Assessment of lipid peroxide levels in plasma Evaluation of antioxidant: status: superoxidedismutase and catalase Evaluation of lipases activity: following parameters will be studied. 1. Free fatty acids. 2. Triglyceride lipase (TGL) activity. 3. Lipoprotein lipase (LPL) activity. 4. Post heparin lipolytic activity (PHLA).

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Some of the above parameters will also be carried out in diabetic patients and patients under the treatment with herbal drugs as well as normal volunteers so that the status of oxidative stress could be compared in such pathological situations.

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Plant Extract Anthocephalus indicus (Kadam): Roots and fruits Hibiscus rosa sinensis (Gurhal): Roots Cassia tora (Chakvat): seeds Tinospora Cordifolia: stems All the plant will be identified by Botanist (CIMAP) Ethanolic extract will be prepare for the treatment

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Anthocephalus indicus (Kadam): Roots and fruits Kadam

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2. Cassia tora (Chakvat): seeds

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Tinospora Cordifolia: stems Gurch

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Hibiscus rosa sinensis (Gurhal): Roots Gurhal

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Study Design Group-1. Healthy Control Group-2. Diabetic Control Group-3. Diabetic + Anthocephalus indicus (Root)- 500mg/kg bw Group-4. Diabetic + Anthocephalus indicus (Fruit) 500mg/kg bw Group-5. Diabetic + Hibiscus rosa sinensis (Root) 500mg/kg bw Group-6. Diabetic + Cassia tora (Seed) 500mg/kg bw Group-7. Diabetic + Tinospora Cordifolia (Stem) 500mg/kg bw Group-7. Diabetic+ Gliblenclamide (600 µ g/ kg b.w.) Following Experimental groups will be treated orally

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Blood Glucose Henry et al ., 1974 . Lipoproteins:  -Lipoprotein (VLDL + LDL) and  -Lipoprotein (HDL) will be precipitated by polyanionic precipitation method of Burstein et al ., 1982 . Total Cholesterol: Zlatkis et al ., 1953. Phospholipids: Fiske and Subbarow, 1952 . Triglyceride: Handel et al ., 1965. Apolipoproteins Separation of apolipoproteins by their selective precipitation with TCA, alcohol and acetone method of Radding & Steinberg, 1960. Lipid-peroxides: Ohkawa et al ., 1979. Free Fatty Acids: Mosinger, 1965 . Triglyceride Lipase (TGL) Activity: Mayes and Felts, 1968. Lipoprotein Lipase (LPL) Activity: Wing and Robinson, 1968 . Superoxide dismutase (SOD) activity: Mc Cord et al ., 1969. Total protein: Lowry et al ., 1951. Catalase activity: Bergmeyer et al ., 1974. Biochemical Parameters

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Importance of the Work The proposed study will be helpful in the advancement of pre existing knowledge and treatment of diabetes. The research work would facilitate in the promotion of knowledge regarding the regulation of different types of dyslipoproteinemias. Studies on dyslipoproteinemia and oxidative stress would further contribute to better understanding in the progression of diabetic complications. This study will lead to the development of antidyslipoproteinemic drugs, from plant extract on animal models. This work will be of great importance due to its significant role in inhibiting the process of micro and macrovascular complications, in diabetes mellitus.

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