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TITLE
DECLARATION
CERTIFICATE
ACKNOWLEDGEMENT
CONTENTS
1. Introduction & Review of Literature.
INTRODUCTION
REVIEW OF LITERATURE
1.1 OXYGEN TOXICITY
1.1.1 Generation of oxygen free radicals and reactive oxygen species
1.1.2 Sources of Reactive Oxygen species
1.1.2.1 Mitochondria
1.1.2.2 Purine metabolism
1.1.2.3 Phagocytes
1.1.2.4 Nitric oxide synthase
1.1.2.5 Transition metals
1.1.2.6 Other endogenous sources
1.1.2.7 Drug metabolism
1.1.2.8 Xenobiotic metabolism
Table 1.1 Free radicals of these compounds implicated in the toxicity to specific tissues.
Bipyridyl herbicides
Fig. 1.4 Redox cycling of chemicals
Halogenated hydrocarbons
Fig. 1.5 Proposed Mechanism of Toxicity of paraquat (Bus and Gitjson, 1984)
Fig. 1.6 Metabolic Activation of CCI4 in rat liver endoplasm reticulum.Slater TF 1987
1.1.2.9 Anti-tumour drugs
Bleomycin
Cisplatin
1.1.2.10 Ionizing Radiation
Fig 1.7 Basic types of structural radiation damage to a cell.
1.1.3 Role Free radicals in diseases processes
1.1.3.1 Free radical - medicated lipid peroxidation and tissue damage.
1.1.3.2 Free radicals in Tumour Promotion
Table 1.2 Free - radical generating compounds that are active as skin tumour promoters.
1.1.3.3 Role of free radicals in carcinogenesis
1.1.3.4 Fibrosis
1.2 RADIOPROTECTION AND CHEMOPROTECTION
1.2.1 Endogenous Antioxidants
Fig. 1.8 The possible actions of potentiators and protectors in modifying free radical induced toxicity and transformation
1.2.1.1 Superoxide dismutases (SOD)
1.2.1.2 Catalase
1.2.1.3 The glutathione System
Glutathione reductase (GR)
1.2.2 Exogenous antioxidants
1.2.2.1 Natural antioxidants
Vitamin E
Vitamin C,
Carotenoids
Polyphenols
Antioxidant chain reaction
1.2.2.2 Synthetic antioxidants
BHA & BHT
Mercaptoalkylamines
Table 1.2 Some Antioxidants available for therapeautic use
1.3 Antioxidants used in this study
1.3.1 α- Tocopherol
1.3.2 Ellagic acid
Fig. 1.10 STRUCTURE OF ANTIOXIDANTS
1.3.3 Curcumin (diferuloyl methane)
1.3.4 Bixin
1.3.5 Probucol (Lorelco)
1.4 Scope of the present study
2. Materials and Methods.
2.1 ANTIOXIDANTS USED IN THE STUDY
2.2 CHEMICALS
2.3 INSTRUMENTS
2.4 ANIMALS
2.5 HUMAN VOLUNTEERS
2.6. DETERMINATION OF THE EFFECT OF ANTIOXIDANlS IN VITRO
2.6.1 Superoxide Scavenging Activity
2.6.2 Hydroxy radical Scavenging Activity
2.6.3 Inhibition of lipid peroxide formation
Induction by Fe 3+/ADP - ascorbate system
2.7 INDUCTION OF LIPID PEROXIDATION IN RATS BY RADIATION
2.8 INDUCTION OF LUNG FIBROSIS IN HATS
2.8.1 Radiation induced lung fibrosis
2.8.2. Bleomycin induced firbosis
2.8.3 Paraquat induced fibrosis
2.9 INDUCTION OF CC14 INDUCED, LIVER FIBROSIS IN RATS
2.10 PREPARATION OF ANTIOXIDANTS FOR ORAL ADMINISTRATION
2.11 EVALUATION OF RADIATION INDUCED GENOTOXICITY IN RATS.
2.11.1 Determination of frequency of Micronuclei induced byRadiation
2.11.2 Detection of Chroraosomal Aberrations Induced by Radiation
2.11.3 Estimation of DNA Strand breaks in Rat lynphocytes
2.12 INDUCTION OF RENAL TOXICITY BY CISPLATIN
2.13 HISTOLOGICAL EXAMINATIONS
2.14 ESTIMATION OF TISSUE LIPID PEROXIDE
2.15 ESTIMATION OF PROTEIN
2.16 ESTIMATION OF SERUM LIPID PEROXIDE
2.17 ESTIMATION OF TISSUE SUPEROXIDE DISMUTASE (S0D)
2.18 ESTIMATION OF TISSUE CATALASE ACTIVITY
2.19 ESllMATlON OF TlSSUE GLUTATHIONE PEROXIDASE
2.20 ESTIMATION OF BLOOD GLUTATHIONE PEROXIDASE (GPX)
2.21 Estimation of Tissue Glutathione (GSB)
2.22 Estimation of cytosolic glutathione- s-traneferase
2.23 ESTIMATION OF CYTOSOLIC GLUTATHIONE REDUCTASE
2.24 ESTIMATION OF TISSUE HYDROXYPROLINE
2.25 ESTIMATION OF TISSUE HYDROXYPROLINE
2.28 ESTIMATION OF GLUTAMATE - PYRUVATE TRANSAMINASE (GPT)
2.29 ESTIMATION OF ALKALINE PHOSPHATASE (ALP) ACTIVITY
2.30 ESTIMATION OF PLASMA UREA
2.31 ESTIMATION OF PLASMA CREATININE
2.32 DETERMINATION OF TOTAL WBC
2.33 STATISTICAL ANALYSIS
3. Comparative Study of the Antioxidant Properties of α-Tocopherol, Ellagic Acid, Curcumin and Bixin.
3.1 MATERIALS AND METHODS
3.1.1 Determination of superoxide scavenging activity.
3.1.2 Determination of hydroxyl radical scavenging activity
3.1.3 Determination of lipid peroxidation inhibiting activity
3.1.4 Effect of α- tocopherol, curcumin and ellagic acid on normal antioxidant defensive mechanism in rats
3.1.5 Biochemical Analysis
3.2 RESULTS
3.2.1 Effect of α-tocopherol, ellagic acid, curcumin and bixin on superoxide generation - in vitro
3.2.2 Effect of α- tocopherol, ellagic acid, curcumin and bixin on hydroxyl radical generation (in vitro)
3.2.3 Effect of α-tocopherol, ellagic acid, curcumin and bixin on lipid peroxidation in vitro
3.2.4 Effect of α-tocopherol, ellagic acid and curcumin on antioxidant defensive mechanism in rats
DISCUSSION
4. Effect of Antioxidants on Radiation Induced Toxicity.
4.1 MATERIALS AND METHODS
4.1.1 EffecL of Lime on radiation induced lipid peroxidation
4.1.2 Effect of radiation dose on lipid peroxidation
4.1.3 Therapeutic dose of radiation on Serum lipid peroxidation
4.1.4 Effect of antioxidants on radiation induced toxicity
4.1.5 Biochemical analyses
4.1.6 Histopathology
4.2 RESULTS
4.2.1 Effect of time on lipid peroxidation
4.2.2 Effect of dose on lipid peroxidation
4.2.3 Effect of therapeutic dose on lipid peroxidation
4.2.4 Effect of antioxidants on radiation induced toxicity.
Fig 4.3 Effect of therapeutic irradiation on serum lipid peroxidation.
4.3 DISCUSSION
5. Effect of Antioxidants on Radiation Induced Genotoxicity.
5.1 MATERIALS AND METHODS
5.1.1 Effect of antioxidants on radiation induced micronuclei formation
5.1.2 Effects of antioxidants on radiation induced chromosomal aberration
5.1.3 Effect of antioxidants on radiation induced DNA trand breaks
5.2 RESULTS
5.2.1 Effect of antioxidants on radiation induced micronuclei formation
5.2.2 Effect of antioxiodants on radiation induced chromosomal aberration
Fig.5.1 Poly chromatic erythrocyte with radiation induced micronuclei
Fig.5.2 Bone marrow cell with radiation induced chromosomal aberration
5.2.3 Effect of antioxidants on radiation induced DNA strand breaks
5.3 DISCUSSION
6. Effect of Antioxidants on Induced Lung Fibrosis.
6.1 MATERIALS AND METHODS
6.1.1 Induction of r-ray induced lung fibrosis
6.1.2 Induction of bleomycin induced lung fibrosis
6.1.3 Induction of paraquat induced lung fibrosis
6.1.4 Biochemical analyses
6.1.5 Histological Examination
6.2 RESULTS
6.2.1 Effect of antioxidants on serum lipidperoxide level
6.2.2 Effect of antioxidants on induced lung lipid peroxidation in rats
6.2.3 Effect of antioxidants on induced liver lipid peroxidation in rats with induced lung fibrosis
6.2.4 Effects of antioxidants of liver superoxide dismutase (SOD) activity
6.2.5 Effect of antioxidants on liver glutathione peroxidaae (GPx) activity
6.2.6 Effect of antioxidants on liver catalase activity
6.2.7 Effect of antioxidants on liver glutathione (GSH) level
6.2.8 The effect of antioxidants on lung collagen hydroxyproline
Fig.6.1 Effect of antioxidant on Radiation induced lung fibrosis in rats
6.2.9 Histological changes
6.3 Discussion
Fig.6.2 Effect of antioxidants on Biomycin induced lung fibrosis in rats
Fig.6.3 Effect of antioxidants on Paraquat induced lung fibrosis in rats
7. Effect of Ellagic Acid on Carbon tetrachloride Induced Liver Fibrosis.
7.1 MATERIALS AND METHODS
7.1.1 Determination of the effect of ellagic acid on liver fibrosis induced by Carbon tetrachloride:
7.1.2 Biochemical Analysis
7.1.3 Histological Examination
7.2 RESULTS
DISCUSSION
8. Effect of α-Tocopherol, Ellagic Acid, Curcumin and Probucol on Cisplatin Induced Renal Toxicity.
8.1 MATERIALS AND METHODS:
8.1.1 Blood Parameters
8.1.2 Tissue parameters.
8.1.3 Histopathological Examination:
8.2 RESULT
8.3 Histopathological observation
8.3 DISCUSSION
9. Summary and Conclusion.
BIBILIOGRAPHY
Papers Published