Chiu Chi Pang. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 121-134). / Abstracts in English and Chinese. / DECLARATION --- p.i / ACKNOWLEDGEMENTS --- p.ii / ABBREVIATIONS --- p.iii / ABSTRACT --- p.vi / 摘要 --- p.viii / Chapter CHAPTER 1: --- GENERAL INTRODUCTION --- p.1 / Chapter 1.1 --- Cholesterol --- p.1 / Chapter 1.1.1 --- History of cholesterol --- p.1 / Chapter 1.1.2 --- Structure of cholesterol --- p.1 / Chapter 1.1.3 --- Biological function of cholesterol --- p.3 / Chapter 1.1.4 --- Sources of cholesterol in our body --- p.3 / Chapter 1.2 --- Lipid hypothesis --- p.4 / Chapter 1.2.1 --- Relationship between dietary cholesterol and plasma cholesterol --- p.4 / Chapter 1.2.2 --- "Hypercholesterolemia , atherosclerosis and coronary heart disease (CHD)" --- p.4 / Chapter 1.2.3 --- Individual variation --- p.5 / Chapter 1.3 --- Cholesterol homeostasis --- p.7 / Chapter 1.3.1 --- SREBPs up-regulates the expression of LDL-receptor and HMG-CoA reductase --- p.7 / Chapter 1.3.2 --- HMG-CoA reductase as the rate-limiting enzyme in cholesterol synthesis --- p.11 / Chapter 1.3.3 --- LDL-receptor as the major protein removing plasma cholesterol …… --- p.12 / Chapter 1.3.4 --- LXR-α as an activator of CYP7A1 --- p.14 / Chapter 1.3.5 --- CYP7A1 controls the classical pathway for the elimination of hepatic cholesterol --- p.16 / Chapter 1.3.6 --- Bile acids as the metabolites of CYP7A1 --- p.17 / Chapter 1.4 --- Previous works in our laboratory --- p.20 / Chapter 1.5 --- Objective of this project --- p.22 / Chapter CHAPTER 2: --- INCREASED EXPRESSION OF LDL-RECEPTOR IS RESPONSIBLE FOR THE HYPO-RESPONSIVENESS OF RATS TO DIETARY CHOLESTEROL --- p.23 / Chapter 2.1 --- Introduction --- p.23 / Chapter 2.2 --- Objective --- p.24 / Chapter 2.3 --- Methods and materials --- p.25 / Chapter 2.3.1 --- Animals --- p.25 / Chapter 2.3.2 --- Diets --- p.25 / Chapter 2.3.3 --- Determination of serum cholesterol --- p.26 / Chapter 2.3.4 --- Western blot --- p.26 / Chapter 2.3.5 --- Probe production for LDL-receptor --- p.27 / Chapter 2.3.5.1 --- Extraction of total RNA --- p.27 / Chapter 2.3.5.2 --- Reverse-transcription reaction of total RNA --- p.28 / Chapter 2.3.5.3 --- Polymerase chain reaction (PCR) of LDL- receptor fragment from cDNA template --- p.28 / Chapter 2.3.5.4 --- Separation and purification of PCR products --- p.29 / Chapter 2.3.5.5 --- Polishing of purified PCR products --- p.29 / Chapter 2.3.5.6 --- Ligation of PCR products and pPCR-script Amp SK(+) cloning vector --- p.30 / Chapter 2.3.5.7 --- Transformation --- p.30 / Chapter 2.3.5.8 --- Preparing glycerol stocks containing the bacterial clones --- p.31 / Chapter 2.3.5.9 --- Plasmid DNA preparation --- p.31 / Chapter 2.3.5.10 --- Clones confirmation by restriction enzyme digestion --- p.32 / Chapter 2.3.5.11 --- Clones confirmation by automatic sequencing --- p.32 / Chapter 2.3.5.12 --- Linearization of the plasmid DNA --- p.33 / Chapter 2.3.5.13 --- DIG-labeling of RNA probe --- p.35 / Chapter 2.3.5.14 --- Testing of DIG-labeled probe --- p.35 / Chapter 2.3.6 --- Probe production for HMG-CoA reductase --- p.36 / Chapter 2.3.7 --- Probe production for GAPDH --- p.37 / Chapter 2.3.8 --- Northern blot --- p.38 / Chapter 2.3.9 --- Determination of hepatic cholesterol --- p.39 / Chapter 2.3.10 --- Statistics --- p.40 / Chapter 2.4 --- Results --- p.42 / Chapter 2.4.1 --- Growth and food intake --- p.42 / Chapter 2.4.2 --- Effect of cholesterol supplements on serum cholesterol --- p.42 / Chapter 2.4.3 --- Effect of cholesterol supplements on liver cholesterol content --- p.45 / Chapter 2.4.4 --- "Stimulatory effect of high cholesterol diet on nSREBP-2, LDL-receptor and HMG-CoA reductase in rats" --- p.45 / Chapter 2.4.5 --- "Effect of high cholesterol diet on nSREBP-2, LDL-receptor and HMG-CoA reductase in hamsters" --- p.49 / Chapter 2.4.6 --- The regulation of LDL-receptor and HMG-CoA reductase existed at transcriptional level --- p.54 / Chapter 2.5 --- Discussion --- p.59 / Chapter CHAPTER 3: --- RATS ARE HYPO-RESPONSIVE TO DIETARY CHOLESTEROL DUE TO EFFICIENT ELIMINATION OF CHOLESTEROL --- p.67 / Chapter 3.1 --- Introduction --- p.67 / Chapter 3.2 --- Objective --- p.69 / Chapter 3.3 --- Methods and materials --- p.70 / Chapter 3.3.1 --- Animals and diets --- p.70 / Chapter 3.3.2 --- Western blot --- p.70 / Chapter 3.3.3 --- Probe production for CYP7A1 and LXR-α --- p.71 / Chapter 3.3.4 --- Northern blot --- p.71 / Chapter 3.3.5 --- Determination of fecal neutral and acidic sterols --- p.71 / Chapter 3.3.5.1 --- Separation of neutral and acidic sterols --- p.71 / Chapter 3.3.5.2 --- Neutral sterols analysis --- p.72 / Chapter 3.3.5.3 --- Acidic sterols analysis --- p.72 / Chapter 3.3.5.4 --- GLC analysis of neutral and acidic sterols --- p.73 / Chapter 3.3.6 --- Statistics --- p.73 / Chapter 3.4 --- Results --- p.76 / Chapter 3.4.1 --- Effect of cholesterol supplements on fecal total neutral sterols --- p.76 / Chapter 3.4.2 --- Effect of cholesterol supplements on fecal total bile acids --- p.76 / Chapter 3.4.3 --- CYP7A1 protein on rats showed a concentration-dependent increase with response to dietary cholesterol while hamsters did not --- p.79 / Chapter 3.4.4 --- The regulation of CYP7A1 was at transcriptional level --- p.79 / Chapter 3.4.5 --- LXR-α demonstrated a parallel changes in its expression at both translational and transcriptional level --- p.84 / Chapter 3.5 --- Discussion --- p.88 / Chapter CHAPTER 4: --- MECHANISM FOR INDIVIDUAL VARIATION OF SERUM CHOLESTEROL LEVEL IN RATS AND HAMSTERS FED A HIGH CHOLESTEROL DIET --- p.94 / Chapter 4.1 --- Introduction --- p.94 / Chapter 4.2 --- Objective --- p.96 / Chapter 4.3 --- Methods and materials --- p.97 / Chapter 4.3.1 --- Diet and animals --- p.97 / Chapter 4.3.2 --- Western blot --- p.97 / Chapter 4.3.3 --- Statistics --- p.97 / Chapter 4.4 --- Results --- p.99 / Chapter 4.4.1 --- Growth and food intake --- p.99 / Chapter 4.4.2 --- Change of serum cholesterol --- p.99 / Chapter 4.4.3 --- Correlation between various protein expression and serum cholesterol --- p.99 / Chapter 4.4.3.1 --- Correlation between LDL-receptor and serum total cholesterol in rats --- p.99 / Chapter 4.4.3.2 --- Correlation between CYP7A1 and serum total cholesterolin rats --- p.99 / Chapter 4.4.3.3 --- Correlation between nSREBP-2 and serum total cholesterolin rats --- p.105 / Chapter 4.4.3.4 --- Correlation between LXR-a and serum total cholesterol in rats --- p.105 / Chapter 4.4.3.5 --- Correlation between HMG-CoA reductase and serum total cholesterol in rats --- p.105 / Chapter 4.4.3.6 --- Correlation between LDL-receptor and serum total cholesterol in hamsters --- p.105 / Chapter 4.4.3.7 --- Correlation between CYP7A1 and serum total cholesterolin hamsters --- p.109 / Chapter 4.4.3.8 --- Correlation between nSREBP-2 and serum total cholesterolin hamsters --- p.109 / Chapter 4.4.3.9 --- Correlation between HMG-CoA reductase and serum total cholesterol in hamsters --- p.109 / Chapter 4.5 --- Discussion --- p.114 / Chapter CHAPTER 5: --- CONCLUSION --- p.117 / REFERENCES --- p.121
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_325294 |
| Date | January 2005 |
| Contributors | Chiu, Chi Pang., 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, ix, 134 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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