Title Page
Abstract
General introduction
Contents
List of abbreviations 16
Chapter 1. Angiotensin II type 1 receptor blocker, Fimasartan, Reduces Cellular Senescence of Coronary Artery Smooth Muscle Cells by inhibiting the CCN1 Signaling Pathway 18
Introduction 19
Materials and methods 22
Results 27
Discussion 44
Chapter 2. Comparison of the Effects of Empagliflozin and Sotagliflozin on a Zebrafish Diabetic Heart Failure with Reduced Ejection Fraction Model 46
Introduction 47
Materials and methods 52
Results 61
Discussion 94
Bibliography 105
Abstract in Korean 116
Chapter 1 12
Figure 1. Angiotensin II-induced cellular senescence in hCASMC and the anti-senescence effect of Fimasartan. (A and B) Counting... 32
Figure 2. Induction of p53 and p16 expression by Angiotensin II and inhibitory effect of Fimasartan. (A-C) Protein and (D and E) mRNA... 33
Figure 3. Induction of gene expression of CCN1 by Angiotensin II and inhibition by Fimasartan. (A) CCN1 mRNA expression following... 34
Figure 4. CCN1 regulates cellular senescence in hCASMCs. (A and B) Counting SA-β-Gal positive cells in hCSMS after Ad-CCN1... 35
Figure 5. Changes in expression of senescence-associated markers p53 and p16 according to CCN1 expression induction. (A, B)... 36
Figure 6. Changes in p53 expression according to CCN1 expression level. (A and B) CCN1 and (C) p53 protein expression level... 37
Figure 7. Changes in expression of senescence-associated markers p53 and p16 according to CCN1 suppression. (A, B) expression... 38
Figure 8. Changes in the expression of senescence-associated markers p53 and p16 following CCN1 inhibition in angiotensin II-... 39
Figure 9. Angiotensin II-induced cellular senescence inhibition by fimasartan via ERK/p38 MAPK/CCN1 signaling pathway. (A and B)... 40
Figure 10. Angiotensin II-induced cellular senescence via ERK/p38 MAPK/CCN1 signaling pathway. (A and B) Changes in... 41
Figure 11. The proposed signaling pathways of Angiotensin II-induced hCASMCs senescence. CCN1 is a molecule that regulates... 43
Chapter 2 13
Figure 1. Effects of various concentrations of D-glucose and/or Streptozotocin on the survival rates and motility of zebrafish larvae.... 72
Figure 2. Induction of hyperglycemia in zebrafish larvae via combined treatment with D-glucose and Streptozotocin. (A)... 73
Figure 3. Induction of diabetes mellitus-like phenotypes in zebrafish larvae via combined treatment with D-glucose and... 74
Figure 4. Reduced cardiac contractility by treatment with Terfenadine in the diabetes mellitus zebrafish model. (A)... 76
Figure 5. Induced irregular cardiac contraction by treatment with Terfenadine in the diabetes mellitus zebrafish model. (A)... 77
Figure 6. Reduced motility and viability in the Terfenadine treated-diabetes mellitus zebrafish model. (A) Representative images of... 78
Figure 7. Comparison of concentration-dependent survival rates of treatment of Empagliflozin or Sotagliflozin. Kaplan-Meier survival... 79
Figure 8. Comparison of concentration-dependent motility conservation effect of Empagliflozin or Sotagliflozin. (A)... 81
Figure 9. Morphology of DM-HFrEF zebrafish treated with Empagliflozin or Sotagliflozin. (A) non-DM, (B) non-DM with... 82
Figure 10. Uninflated swim bladder in high dose sotagliflozin treated HF induced non-DM zebrafish model. (A) Morphology of non-DM... 83
Figure 11. Cardiac contractile functions in the DM-HFrEF zebrafish model. (A) Representative fluorescent microscopy images of Tg... 84
Figure 12. Treatment with Empagliflozin or Sotagliflozin improved cardiac contractility in the DM-HFrEF zebrafish model. (A)... 86
Figure 13. Treatment with Empagliflozin or Sotagliflozin suppress irregular contraction in the DM-HFrEF zebrafish model. (A)... 87
Figure 14. Gene expression of nppb, ins and pck1 in the DM-HFrEF zebrafish treated with Empagliflozin or Sotagliflozin. (A) Relative... 88
Figure 15. Empagliflozin and Sotagliflozin structurally bind to zebrafish NHE1 in silico. (A and B) Molecular docking analysis of... 89
Figure 16. Empagliflozin and Sotagliflozin structurally bind to zebrafish NHE1 in vitro. (A) Schematic of the drug affinity... 90
Figure 17. Empagliflozin and sotagliflozin functionally inhibit zebrafish NHE1 in vitro. (A) Measurement of intracellular Na+, (B)...[이미지참조] 91
Figure 18. Inhibition of intracellular ion changes by Empagliflozin or Sotagliflozin in a concentration-dependent manner. (A) Intracellular... 92
Figure 19. Competitive Inhibitory Effects of Cariporide and Empagliflozin or Sotagliflozin. (A) intracellular Na+ and (B) Ca2+... 93