Title Page
Abstract
Contents
Chapter 1. Introduction 11
1.1. Chronic Kidney Disease Accelerates Atherosclerosis Causing Cardiovascular Diseases 11
1.2. Mechanism of Accelerated Atherosclerosis in CKD 11
1.3. Atorvastatin Attenuates Atherosclerosis 12
1.4. Protective Effect of mTOR Inhibitor (Rapamycin) Against Atherosclerosis 12
1.5. Combination of Atorvastatin and Rapamycin 13
1.6. Purpose of the Study 13
Chapter 2. Materials and Methods 15
2.1. Experimentally Induced CKD Mice Model (Two-Step Procedure) 15
2.2. Blood Biochemistry 16
2.3. Atherosclerotic Lesion Analysis 17
2.4. RNA Isolation and Quantitative Real-Time PCR 19
2.5. Western Blot Analysis 21
2.6. Fast Protein Liquid Chromatography (FPLC) Analysis 21
2.7. Immunostaining 22
2.8. Statistical Analysis 22
Chapter 3. Results 23
3.1. Rapamycin Ameliorates CKD-Associated Atherosclerosis 23
3.2. Combination of Rapamycin and Atorvastatin Improves Lipid Metabolism in CKD Mice 28
3.3. ApoE-/- Mice with CKD Exhibit Up-Regulation of Pro-Inflammatory Cytokine Genes in the Spleen and Aorta, and these Levels Are Reduced by Rapamycin 34
Chapter 4. Discussion 39
4.1. Experimentally Induced CRF Mice Model (Two-Step Procedure) 39
4.2. Combination of Rapamycin and Atorvastatin Improves Lipid Metabolism 40
4.3. Rapamycin And Atorvastatin Markedly Decreased the Gene Expression Levels of Atherosclerosis-Promoting Inflammatory 41
4.4. Atorvastatin in CKD 42
4.5. Conclusion 42
Bibliography 44
국문 초록 52
Table 1. Primer Sequences 20
Table 2. Body Weight and Laboratory Data 25
Figure 1. (A) Schematic presentation of the experimental protocol. (B) Two weeks later, left total nephrectomy was performed through a left flank incision.... 16
Figure 2. Aorta was perfused through the left ventricle with ice-cold phosphate-buffered saline (A) and was opened along the longitudinal axis and... 18
Figure 3. Body weight and levels of blood urea nitrogen and creatinine 24
Figure 4. Fast Protein Liquid Chromatography (FPLC) fraction cholesterol levels from pooled plasma samples. The CKD group showed a marked increase in VLDL. 26
Figure 5. Rapa ameliorates the formation of atherosclerotic plaques, as assessed by Oil red O staining. (A) Representative en face images of whole... 27
Figure 6. Sections of the aortic sinus were stained with the macrophage marker, Moma-2 (red), to determine macrophage content per plaque size. SM α-actin... 28
Figure 7. Combined treatment with Rapa plus Atv has beneficial effects on lipid metabolism. Serum lipid profiles for (A) total cholesterol, (B) triglycerides, (D)... 30
Figure 8. mRNA expression levels of (A) cholesterol metabolism-related genes, (B, C) cholesterol transport-related genes, (D) bile acid biosynthesis genes, and... 31
Figure 9. The expression levels of reverse cholesterol transport-related proteins in the livers of the indicated groups (n=5 per group). Data are shown... 33
Figure 10. The effects of RAPA and ATV on the CKD-related stimulation of pro atherosclerosis cytokines in ApoE-/- mice. Gene expression levels of... 35
Figure 11. The effects of Rapa and Atv on the CKD-related stimulation of pro atherosclerosis cytokines in ApoE-/- mice. Gene expression levels of... 37