by Yau Ming Hon. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 131-154). / Abstracts in English and Chinese. / Contents --- p.i / Acknowledgements --- p.ix / Abstract --- p.x / 槪論 --- p.xi / List of abbreviations --- p.xii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Reactive oxygen species (ROS) --- p.2 / Chapter 1.1.1 --- Hydrogen peroxide --- p.2 / Chapter 1.1.2 --- Superoxide anion --- p.3 / Chapter 1.1.3 --- Hydroxyl radical --- p.3 / Chapter 1.1.4 --- Carbon centered radical --- p.4 / Chapter 1.1.5 --- Nitric oxide --- p.4 / Chapter 1.2 --- Physiological roles of ROS --- p.5 / Chapter 1.2.1 --- Signal transduction --- p.5 / Chapter 1.2.2 --- Phagocytic activity --- p.6 / Chapter 1.3 --- Defense systems against ROS --- p.7 / Chapter 1.3.1 --- Endogenous antioxidant enzymes --- p.8 / Chapter 1.3.1.1 --- Catalase --- p.8 / Chapter 1.3.1.2 --- Superoxide dismutase --- p.8 / Chapter 1.3.1.3 --- Selenium-glutathione peroxidase --- p.9 / Chapter 1.3.1.4 --- Glutathione reductase --- p.10 / Chapter 1.3.1.5 --- Glutathione-S-transferases --- p.10 / Chapter 1.3.2 --- Non-enzymatic antioxidants --- p.12 / Chapter 1.3.2.1 --- Vitamin E (tocopherols and tocotrienols) --- p.12 / Chapter 1.3.2.2 --- Vitamin C (L-ascorbic acid) --- p.13 / Chapter 1.3.2.3 --- Glutathione --- p.14 / Chapter 1.3.2.4 --- Flavonoids and polyphenols --- p.15 / Chapter 1.3.2.5 --- Uric acid --- p.16 / Chapter 1.4 --- Roles of ROS in pathogenesis --- p.16 / Chapter 1.4.1 --- Liver diseases --- p.17 / Chapter 1.4.2 --- Genital malfunctioning --- p.19 / Chapter 1.5 --- "The traditional Chinese medicinal formula, Wu-zi-yan-zong-wan" --- p.19 / Chapter 1.5.1 --- Pharmacology of individual herbal components --- p.20 / Chapter 1.5.1.1 --- Semen Cuscuta --- p.20 / Chapter 1.5.1.2 --- Fructus Lycii --- p.21 / Chapter 1.5.1.3 --- Fructus Schisandrae --- p.21 / Chapter 1.5.1.4 --- Fructus Rubi --- p.22 / Chapter 1.5.1.5 --- Semen Plantaginis --- p.22 / Chapter 1.5.2 --- Effect of Wu-zi-yan-zong-wan on infertility --- p.23 / Chapter 1.5.3 --- Effect of Wu-zi-yan-zong-wan on liver disease --- p.23 / Chapter 1.6 --- Objectives of the present study --- p.24 / Chapter Chapter 2 --- Antioxidant Activity of Aqueous Extracts of the Herbal Components of Wu-zi-yan-zong-wan in in vitro Free Radical Generating Systems --- p.26 / Chapter 2.1 --- Introduction --- p.27 / Chapter 2.1.1 --- Application of in vitro ROS generating systems --- p.27 / Chapter 2.1.1.1 --- Superoxide generation --- p.27 / Chapter 2.1.1.2 --- Hydroxyl radical generation system --- p.28 / Chapter 2.1.1.3 --- "2,2'-Azobis(2-amidinopropane) dihydrochloride- induced hemolysis" --- p.28 / Chapter 2.1.1.4 --- Bleomycin-iron-dependent DNA damage --- p.28 / Chapter 2.1.2 --- Objective --- p.29 / Chapter 2.2 --- Materials and methods --- p.30 / Chapter 2.2.1 --- Materials --- p.30 / Chapter 2.2.2 --- Preparation of aqueous herbal extracts --- p.30 / Chapter 2.2.3 --- Superoxide-scavenging assay --- p.30 / Chapter 2.2.4 --- Microsome lipid peroxidation inhibition assay --- p.31 / Chapter 2.2.5 --- "2,2'-Azobis(2-amidinopropane) dihydrochloride-induced hemolysis inhibition assay" --- p.32 / Chapter 2.2.6 --- Bleomycin-iron-dependent DNA damage inhibition assay --- p.32 / Chapter 2.2.7 --- Statistical analysis --- p.33 / Chapter 2.3 --- Results --- p.34 / Chapter 2.3.1 --- Extraction yield --- p.34 / Chapter 2.3.2 --- Free radical scavenging activity of Wu-zi-yan-zong-wan extract --- p.34 / Chapter 2.3.3 --- Free radical scavenging activity of FR extract --- p.37 / Chapter 2.3.3.1 --- Superoxide-scavenging activity --- p.37 / Chapter 2.3.3.2 --- Effect on hydroxyl radical-induced lipid peroxidation --- p.37 / Chapter 2.3.3.3 --- Effect on AAPH-induced hemolysis --- p.40 / Chapter 2.3.3.4 --- Effect on bleomycin-iron-dependent DNA damage --- p.40 / Chapter 2.3.4 --- Pro-oxidant activity of FR extract --- p.40 / Chapter 2.3.5 --- Free radical scavenging activity of the remaining herbal extracts --- p.44 / Chapter 2.4 --- Discussion --- p.46 / Chapter Chapter 3 --- Effect of Aqueous Extract of the Herbal Components of Wu- zi-yan-zong-wan on tert-Butyl Hydroperoxide-Induced Oxidative Damage in Primary Rat Hepatocyte --- p.51 / Chapter 3.1 --- Introduction --- p.52 / Chapter 3.1.1 --- Primary rat hepatocyte as pharmacological model --- p.52 / Chapter 3.1.2 --- tert-Butyl hydroperoxide as an oxidative stress inducer --- p.53 / Chapter 3.1.3 --- Detection of ROS --- p.54 / Chapter 3.1.4 --- Objective --- p.55 / Chapter 3.2 --- Materials and methods --- p.56 / Chapter 3.2.1 --- Materials --- p.56 / Chapter 3.2.2 --- Primary rat hepatocyte isolation --- p.56 / Chapter 3.2.2.1 --- Liver perfusion --- p.56 / Chapter 3.2.2.2 --- Collagen pre-coated plates preparation --- p.57 / Chapter 3.2.2.3 --- Hepatocyte culture --- p.58 / Chapter 3.2.3 --- Drug treatment and oxidative stress induction --- p.58 / Chapter 3.2.4 --- Cytotoxicity assessment --- p.58 / Chapter 3.2.4.1 --- Lactate dehydrogenase leakage measurement --- p.59 / Chapter 3.2.4.2 --- MTT assay --- p.59 / Chapter 3.2.5 --- Cellular GSH content determination --- p.59 / Chapter 3.2.6 --- Protein determination by Lowry's method --- p.60 / Chapter 3.2.7 --- MDA measurement --- p.60 / Chapter 3.2.8 --- GSSG measurement --- p.61 / Chapter 3.2.9 --- ROS measurement with fluorescent dye --- p.61 / Chapter 3.2.10 --- "Vitamin C, vitamin E and butylated hydroxytoluene treatment" --- p.62 / Chapter 3.2.11 --- Antioxidant enzyme activity measurement --- p.62 / Chapter 3.2.11.1 --- Catalase activity measurement --- p.62 / Chapter 3.2.11.2 --- Superoxide dismutase activity measurement --- p.63 / Chapter 3.2.11.3 --- Glutathione peroxidase activity measurement --- p.63 / Chapter 3.2.11.4 --- Glutathione-S-transferases activity measurement --- p.63 / Chapter 3.2.11.5 --- Glutathione reductase activity measurement --- p.64 / Chapter 3.2.12 --- Statistical analysis --- p.64 / Chapter 3.3 --- Results --- p.65 / Chapter 3.3.1 --- Cytotoxicity of FR extract on rat hepatocyte --- p.65 / Chapter 3.3.2 --- Effect of tBHP and FR extract on hepatocyte viability --- p.65 / Chapter 3.3.3 --- Time-dependent effect of FR extract on tBHP-induced cytotoxicity --- p.69 / Chapter 3.3.4 --- Effect of tBHP and FR extract on hepatocyte GSH content --- p.69 / Chapter 3.3.5 --- Effect of tBHP and FR extract on GSSG formation in hepatocyte --- p.72 / Chapter 3.3.6 --- Effect of tBHP and FR extract on MDA formation in hepatocyte --- p.72 / Chapter 3.3.7 --- ROS-scavenging activity of FR extract in hepatocyte --- p.77 / Chapter 3.3.8 --- Effect of FR extract on antioxidant enzymes activities --- p.77 / Chapter 3.3.9 --- Comparison between typical antioxidants --- p.77 / Chapter 3.3.10 --- Effect of WZ and remaining herbal extracts on tBHP-induced oxidative damage in hepatocyte --- p.81 / Chapter 3.4 --- Discussion --- p.84 / Chapter Chapter 4 --- Effect of Aqueous Extract of Wu-zi-yan-zong-wan and Fructus Rubi on tert-Buty Hydroperoxide Induced Oxidative Damage in Mouse Model --- p.91 / Chapter 4.1 --- Introduction --- p.92 / Chapter 4.2 --- Materials and methods --- p.93 / Chapter 4.2.1 --- Materials --- p.93 / Chapter 4.2.2 --- Animal treatments --- p.93 / Chapter 4.2.3 --- Serum preparation --- p.94 / Chapter 4.2.4 --- Marker enzyme measurement --- p.94 / Chapter 4.2.5 --- Liver MDA and GSH determination --- p.95 / Chapter 4.2.6 --- Statistical analysis --- p.95 / Chapter 4.3 --- Results --- p.97 / Chapter 4.3.1 --- Effect of tBHP and FR extract on mouse serum ALT and AST activities --- p.97 / Chapter 4.3.2 --- Effect of tBHP and FR extract on mouse liver MDA and GSH content --- p.97 / Chapter 4.3.3 --- Effect of WZ extract on tBHP-induced increase in serum ALT and AST activities --- p.97 / Chapter 4.4 --- Discussion --- p.102 / Chapter Chapter 5 --- Characterization of the Active Antioxidant Principlein Aqueous Extract of FR --- p.105 / Chapter 5.1 --- Introduction --- p.106 / Chapter 5.2 --- Materials and methods --- p.107 / Chapter 5.2.1 --- Materials --- p.107 / Chapter 5.2.2 --- Chemical/physical treatments on FR extract --- p.107 / Chapter 5.2.3 --- Digestion with enzymes --- p.108 / Chapter 5.2.4 --- Antioxidant activity determination --- p.109 / Chapter 5.2.5 --- Chemical composition determination --- p.109 / Chapter 5.2.5.1 --- Uronic acid determination --- p.109 / Chapter 5.2.5.2 --- Hexose determination --- p.109 / Chapter 5.2.5.3 --- Tannin determination --- p.110 / Chapter 5.2.5.4 --- Protein determination --- p.110 / Chapter 5.2.6 --- Column chromatography --- p.110 / Chapter 5.2.6.1 --- Polyamide CC6 resin column chromatography --- p.111 / Chapter 5.2.6.2 --- Sephadex LH-20 gel column chromatography --- p.111 / Chapter 5.2.7 --- Antioxidant activity of commercially available tannin --- p.111 / Chapter 5.2.8 --- Bovine serum albumin precipitation --- p.112 / Chapter 5.2.9 --- Statistical analysis --- p.112 / Chapter 5.3 --- Results --- p.113 / Chapter 5.3.1 --- Effect of chemical/physical treatments on antioxidant activity of FR extract --- p.113 / Chapter 5.3.2 --- Effect of enzyme digestions on antioxidant activity of FR extract --- p.113 / Chapter 5.3.3 --- Chemical composition of FR extract --- p.118 / Chapter 5.3.4 --- Polyamide CC6 resin column chromatography --- p.118 / Chapter 5.3.5 --- Sephadex LH-20 gel column chromatography --- p.118 / Chapter 5.3.6 --- Antioxidant activity of commercially available tannin --- p.123 / Chapter 5.3.7 --- Effect of BSA precipitation on superoxide-scavenging activity --- p.123 / Chapter 5.4 --- Discussion --- p.127 / Conclusion --- p.131 / References --- p.132
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324068 |
| Date | January 2002 |
| Contributors | Yau, Ming Hon., Chinese University of Hong Kong Graduate School. Division of Biochemistry. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xiii, 154 leaves : ill. ; 30 cm. |
| Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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