ABSTRACT
Macrophage foam cell formation is the critical step in the development of atherosclerosis. Berberine, a botanical alkaloid, has been suggested to regulate lipid homeostasis by reducing serum cholesterol and triglyceride levels and to improve glucose metabolism. This study investigated the role of Berberine on lipid metabolism in macrophages. In THP-1-derived macrophages, Berberine suppressed lipid accumulation induced by oxidized-low density lipoprotein (ox-LDL). Berberine augmented the mRNA and protein expression of ATP-binding membrane cassette transport protein A1 (ABCA1) but did not alter the protein levels of ATP-binding membrane cassette transport protein G1 (ABCG1)、scavenger receptor B1 (SR-B1)、scavenger receptor A (SR-A) and clusters of differentiation 36 (CD36). Moreover, ABCA1 antibody or inhibitor abrogated the suppressive effect of Berberine on lipid accumulation. Finally, western blot analysis and confocal microscopy revealed that Berberine enhanced the nuclear translocation of liver X receptor-a□ but not its protein expression. These data suggest that Berberine inhibits the formation of foam cells by enhancing cholesterol efflux and could be effective for treatment of atherosclerosis.

Table of Contents

誌謝 I
中文摘要 VI
Abstract VI

Table of Contents VII
List of Abbreviations XI
List of Illustrations XII
List of Tables XIV
List of Appendixes XV

Chapter 1. General Introduction 1
1.1 Cardiovascular Diseases and Atherosclerosis 1
1.1.1 Epidemiology 1
1.1.2 Atherosclerosis ~ the General Description 2
1.1.3 Hypotheses of Atherogenesis 3
1.1.3.1 The Response-to-Injury Hypothesis 4
1.1.3.2 The Response-to-Retention Hypothesis 4
1.1.3.3 the Oxidative Modification Hypothesis 5
1.2 Foam Cell Formation 7
1.2.1 Lipoprotein and Macrophage Differentiation 7
1.2.2 Macrophage Inflammation and Foam Cell Formation 10
1.2.2.1 Toll-like receptors and inflammation 11
1.2.2.2 Scavenger receptors and cholesterol accumulation 11
1.2.2.3 Macrophage Foam Cell Formation 12
1.3 Reverse Cholesterol Transport and Cholesterol Efflux 13
1.3.1 Reverse Cholesterol Transport 13
1.3.2 Cholesterol Efflux of Macrophages 15
1.4 ABCA1 ~ the Major Cholesterol Efflux Transporter 16
1.4.1 Structure and Function of ABCA1 16
1.4.2 Substrates and Lipid Acceptors for ABCA1 17
1.5 LXR-mediated ABCA1 Transcriptional Regulation is a
Potential Therapeutic Target 19
1.5.1 ABCA1 as a Protector from Cardiovascular Diseases 19
1.5.2 LXRs as Integrators of Metabolic Signaling 20
1.6 Berberine ~ a Potential Drug in Treatment of Atherosclerosis 24
1.7 Research Motivation 26
Chapter 2. Materials and Methods 29
2.1 Reagents 29
2.2 Methods 30
2.2.1 Cell Culture 30
2.2.2 Oxidation of LDL 30
2.2.3 Foam Cell Formation 31
2.2.4 Oil red O staining for Cytoplasmic Lipid Detection 31
2.2.4.1 Preparation of Oil Red O staining solution 31
2.2.4.2 Oil red O staining for microscopy examination 32
2.2.4.3 Quantification of the cytoplasmic lipid detection¬ 32
2.2.5 Preparation of Cytosolic Fraction and Nuclear Extracts 32
2.2.6 Preparation of whole Cell Extracts 33
2.2.7 Western Blot Analysis 33
2.2.8 Reverse Transcription-Polymerase Chain Reaction 34
2.2.9 Immunofluorescence Assay 35
2.2.10 Transfection 35
2.2.11 Luciferase Reporter Assay 36
2.2.12 Statistical Analysis 36
Chapter 3. Experimental Results 37
3.1 Berberine Attenuates oxLDL-Induced Lipid Accumulation
in Macrophage 37
3.2 Effects of Berberine on Expression of RCTs and SRs
in THP-1-derived Macrophages 40
3.3 Inhibition of ABCA1 Abrogates the Lipid-Lowering Effect
of Berberine 46
3.4 Berberine Induces Nuclear Translocation of LXR 48
Chapter 4. Discussion 55
References 58
Appendix 75

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