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Effects of octadecaenoic acids and apple polyphenols on blood cholesterol.

Lam, Cheuk Kai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 148-173). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABSTRACT --- p.ii / LIST OF ABBREVIATIONS --- p.vi / TABLE OF CONTENTS --- p.x / Chapter CHAPTER 1 --- GENERAL INTRODUCTION / Chapter 1.1 --- Introduction to Cholesterol and Its Related Diseases --- p.1 / Chapter 1.1.1 --- Chemistry of cholesterol --- p.1 / Chapter 1.1.2 --- Physiological importance of cholesterol --- p.1 / Chapter 1.1.3 --- Pathological effects of cholesterol --- p.3 / Chapter 1.1.3.1 --- Mechanism of atherosclerosis --- p.3 / Chapter 1.2 --- Cholesterol Homeostasis --- p.6 / Chapter 1.2.1 --- Liver as the main organ for cholesterol metabolism --- p.6 / Chapter 1.2.2 --- Regulatory sites of cholesterol metabolism --- p.6 / Chapter 1.2.2.1 --- Regulation of cholesterol absorption by acyl coenzyme A: cholesterol acyltransferase (ACAT) --- p.6 / Chapter 1.2.2.2 --- Sterol regulatory element-binding protein 2 (SREBP-2) as a transcription factor for 3 -hydroxy-3 -methylglutaryl coenzyme A reductase (HMGR) and low-density lipoprotein receptor (LDLR) --- p.10 / Chapter 1.2.2.3 --- Roles ofLDLR --- p.11 / Chapter 1.2.2.4 --- Rate limiting role of HMGR in cholesterol de novo synthesis --- p.14 / Chapter 1.2.2.5 --- Roles of liver-X-receptor-a (LXR-a) in cholesterol catabolism --- p.16 / Chapter 1.2.2.6 --- Roles of CYP7A1 in catabolism of cholesterol into bile acids --- p.19 / Chapter 1.2.2.7 --- Roles of cholesterol ester transfer protein (CETP) in maintaining cholesterol distribution in blood --- p.22 / Chapter CHAPTER 2 --- EFFECT OF OCTADECAENOIC ACIDS ON BLOOD CHOLESTEROL IN HAMSTERS / Chapter 2.1 --- Introduction --- p.25 / Chapter 2.1.1 --- Effects of polyunsaturated fatty acids (PUFAs) on blood cholesterol --- p.25 / Chapter 2.1.2 --- Differential effects of 18-C PUFAs on lowering blood cholesterol in vivo --- p.25 / Chapter 2.1.3 --- "Structures, metabolism and conjugation of octadecaenoic acids (ODA)" --- p.26 / Chapter 2.1.4 --- Objectives --- p.26 / Chapter 2.2 --- Experiment 1 --- p.28 / Chapter 2.2.1 --- Materials and methods --- p.28 / Chapter 2.2.1.1 --- Experimental fatty acids --- p.28 / Chapter 2.2.1.1.1 --- Isolation of LN from flaxseed --- p.28 / Chapter 2.2.1.1.2 --- Isolation of CLN from tung seed --- p.28 / Chapter 2.2.1.2 --- Animals --- p.29 / Chapter 2.2.1.3 --- Diets --- p.30 / Chapter 2.2.1.4 --- Plasma lipid measurements --- p.30 / Chapter 2.2.1.5 --- Plasma CETP activity measurement --- p.30 / Chapter 2.2.1.6 --- "Measurement of liver SREBP-2, LDLR, HMGR and CYP7A1 protein abundance by Western blotting" --- p.34 / Chapter 2.2.1.7 --- "Measurement of hepatic SREBP-2, LDLR, HMGR, LXR, CYP7A1, CETP, SR-B1 and LCAT mRNA by real time PCR" --- p.35 / Chapter 2.2.1.7.1 --- Extraction of mRNA --- p.35 / Chapter 2.2.1.1.2 --- Complementary DNA synthesis --- p.36 / Chapter 2.2.1.7.3 --- Real-time polymerase chain reaction (PCR) anaylsis --- p.36 / Chapter 2.2.1.8 --- Determination of cholesterol in liver --- p.37 / Chapter 2.2.1.9 --- Determination of fecal neutral and acidic sterols --- p.38 / Chapter 2.2.1.9.1 --- Determination of fecal neutral sterols --- p.39 / Chapter 2.2.1.9.2 --- Determination of fecal acidic sterols --- p.41 / Chapter 2.2.1.10 --- Statistics --- p.43 / Chapter 2.2.2 --- Results --- p.44 / Chapter 2.2.2.1 --- Growth and food intake --- p.44 / Chapter 2.2.2.2 --- Organ weights --- p.44 / Chapter 2.2.2.3 --- "Effects of ODA on serum TC, TG and HDL-C" --- p.44 / Chapter 2.2.2.4 --- Effect of ODA on liver cholesterol --- p.48 / Chapter 2.2.2.5 --- Effect of ODA on fecal neutral sterol output --- p.48 / Chapter 2.2.2.6 --- Effect of ODA on fecal acidic sterol output --- p.48 / Chapter 2.2.2.7 --- Effect of ODA on cholesterol balance in hamsters --- p.52 / Chapter 2.2.2.8 --- Effect of ODA on plasma CETP activity --- p.52 / Chapter 2.2.2.9 --- Correlation between blood TC and liver cholesterol --- p.52 / Chapter 2.2.2.10 --- Correlation between blood HDL-C and liver cholesterol --- p.52 / Chapter 2.2.2.11 --- Correlation between blood nHDL/HDL ratio and liver cholesterol --- p.52 / Chapter 2.2.2.12 --- Effect ofODA on liver SREBP-2 immunoreactive mass --- p.58 / Chapter 2.2.2.13 --- Effect of ODA on liver LDLR immunoreactive mass --- p.58 / Chapter 2.2.2.14 --- Effect of ODA on liver HMGR immunoreactive mass --- p.58 / Chapter 2.2.2.15 --- Effect of ODA on liver LXR immunoreactive mass --- p.58 / Chapter 2.2.2.16 --- Effect of ODA on liver CYP7A1 immunoreactive mass --- p.63 / Chapter 2.2.2.17 --- Effects ofODA on hepatic CETP mRNA --- p.65 / Chapter 2.2.2.18 --- Effects of ODA on hepatic LDLR mRNA --- p.65 / Chapter 2.2.2.19 --- Effects of ODA on hepatic LXR mRNA --- p.65 / Chapter 2.2.2.20 --- Effects of ODA on hepatic CYP7A1 mRNA --- p.65 / Chapter 2.3 --- Experiment 2 --- p.70 / Chapter 2.3.1 --- Materials and Methods --- p.70 / Chapter 2.3.1.1 --- Experimental diets --- p.70 / Chapter 2.3.1.2 --- Animals --- p.70 / Chapter 2.3.1.3 --- Intestinal acyl coenzyme A: cholesterol acyltransferase (ACAT) activity measurement --- p.70 / Chapter 2.3.1.3.1 --- Preparation of intestinal microsome --- p.71 / Chapter 2.3.1.3.2 --- ACAT activity assay --- p.71 / Chapter 2.3.2 --- Results --- p.73 / Chapter 2.3.2.1 --- Growth and food intake --- p.73 / Chapter 2.3.2.2 --- Organ weights --- p.73 / Chapter 2.3.2.3 --- "Effect of ODA on serum TC, TG and HDL-C" --- p.73 / Chapter 2.3.2.4 --- Effect of ODA feeding on fecal neutral sterol content --- p.77 / Chapter 2.3.2.5 --- Effect of ODA feeding on fecal acidic sterol content --- p.77 / Chapter 2.3.2.6 --- Effect of ODA feeding on intestinal acyl coenzyme A: acyl cholesterol transferase (ACAT) activity --- p.77 / Chapter 2.4 --- Discussion --- p.81 / Chapter CHAPTER 3 --- EFFECT OF OCTADECAENOIC ACIDS ON CHOLESTEROL-REGULATING GENES IN HepG2 / Chapter 3.1 --- Introduction --- p.86 / Chapter 3.1.1 --- HepG2 as a model of cholesterol regulation --- p.86 / Chapter 3.1.2 --- Effect of polyunsaturated fatty acids (PUFAs) on cholesterol regulating genes in cultured cells --- p.87 / Chapter 3.1.3 --- Objectives --- p.89 / Chapter 3.2 --- Materials and Methods --- p.90 / Chapter 3.2.1 --- Cell culture --- p.90 / Chapter 3.2.2 --- "Measurement of SREBP-2, LDLR, HMGR and CYP7A1 protein abundance by Western blotting" --- p.92 / Chapter 3.2.3 --- "Measurement of cellular SREBP-2, LDLR, HMGR, LXR, CYP7A1 and CETP mRNA by real time PCR" --- p.93 / Chapter 3.2.4 --- Statistics --- p.93 / Chapter 3.3 --- Results --- p.95 / Chapter 3.3.1 --- Effect of ODA on HepG2 SREBP-2 immunoreactive mass --- p.95 / Chapter 3.3.2 --- Effect of ODA on HepG2 HMGR immunoreactive mass --- p.95 / Chapter 3.3.3 --- Effect of ODA on HepG2 LDLR immunoreactive mass --- p.95 / Chapter 3.3.4 --- Effect of ODA on HepG2 LXR immunoreactive mass --- p.95 / Chapter 3.3.5 --- Effect of ODA on HepG2 CYP7A1 immunoreactive mass --- p.96 / Chapter 3.3.6 --- Effect of ODA supplementation on HepG2 SREBP-2 mRNA expression --- p.102 / Chapter 3.3.7 --- Effect of ODA supplementation on HepG2 SREBP-2 mRNA expression --- p.102 / Chapter 3.3.8 --- Effect of ODA supplementation on HepG2 LDLR mRNA expression --- p.102 / Chapter 3.3.9 --- Effect of ODA supplementation on HepG2 LXR mRNA expression --- p.106 / Chapter 3.3.10 --- Effect of ODA supplementation on HepG2 CYP7A1 mRNA expression --- p.106 / Chapter 3.3.11 --- Effect of ODA supplementation on HepG2 CETP mRNA expression --- p.106 / Chapter 3.4 --- Discussion --- p.110 / Chapter CHAPTER 4 --- EFFECT OF APPLE POLYPHENOLS ON BLOOD CHOLESTEROL IN HAMSTERS / Chapter 4.1 --- Introduction --- p.114 / Chapter 4.1.1 --- Apple is a commonly consumed fruit worldwide --- p.114 / Chapter 4.1.2 --- Potential health effects of apples --- p.114 / Chapter 4.1.3 --- Abundance of polyphenols in apple --- p.115 / Chapter 4.1.4 --- Fuji variety of apple --- p.116 / Chapter 4.1.5 --- Objectives --- p.116 / Chapter 4.2 --- Materials and Methods --- p.118 / Chapter 4.2.1 --- Isolation of AP --- p.118 / Chapter 4.2.2 --- Characterization of AP extract --- p.118 / Chapter 4.2.3 --- Effect of AP on CETP activity in vitro --- p.118 / Chapter 4.2.4 --- Effect of AP on blood cholesterol in hamsters --- p.119 / Chapter 4.2.4.1 --- Animals --- p.119 / Chapter 4.2.4.2 --- Diets --- p.120 / Chapter 4.2.4.3 --- Plasma lipids measurement --- p.121 / Chapter 4.2.4.4 --- Plasma CETP activity measurement and immunoreactive mass by Western blotting --- p.123 / Chapter 4.2.4.5 --- "Measurement of liver SREBP-2, LDL-R, HMG-R and CYP7A1 protein abundance by Western blotting" --- p.124 / Chapter 4.2.4.6 --- Statistics --- p.124 / Chapter 4.3 --- Results --- p.125 / Chapter 4.3.1 --- Polyphenol content in AP --- p.125 / Chapter 4.3.2 --- Effect of AP on CETP activity in vitro --- p.125 / Chapter 4.3.3 --- Growth and food intake --- p.128 / Chapter 4.3.4 --- Organ weights --- p.128 / Chapter 4.3.5 --- Effect of AP supplementation on the plasma lipid profile of hamsters --- p.131 / Chapter 4.3.6 --- Effect of AP feeding on plasma CETP activity of the hamsters --- p.131 / Chapter 4.3.7 --- Effect of AP on plasma CETP immunoreactive mass --- p.134 / Chapter 4.3.8 --- Effect of AP on liver SREBP-2 immunoreactive mass --- p.134 / Chapter 4.3.9 --- Effect of AP on liver LDLR immunoreactive mass --- p.134 / Chapter 4.3.10 --- Effect of AP on liver HMGR immunoreactive mass --- p.134 / Chapter 4.3.11 --- Effect of AP on liver CYP7A1 immunoreactive mass --- p.134 / Chapter 4.3.12 --- Effect of AP on liver cholesterol level --- p.140 / Chapter 4.4 --- Discussion --- p.142 / Chapter CHAPTER 5 --- CONCLUSION --- p.145 / REFERENCES --- p.148

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326080
Date January 2007
ContributorsLam, Cheuk Kai., Chinese University of Hong Kong Graduate School. Division of Food and Nutritional Sciences.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xvi, 173 leaves : ill. ; 30 cm.
RightsUse 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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