Title Page
Contents
Abbreviations 11
국문초록 13
ABSTRACT 16
Ⅰ. INTRODUCTION 20
A. Diabetic nephropathy 20
B. Echinochrome A 26
Ⅱ. PURPOSE OF THE STUDY 29
Ⅲ. MATERIALS AND METHODS 30
A. Experimental animals and drug treatments 30
1. Echinochrome A Preparation 30
2. Experimental animals 31
B. Fasting Blood Glucose and Intraperitoneal Glucose Tolerance Test 32
C. Measurement of Blood Creatinine, BUN, and Insulin Levels 33
D. Histological Analyses 33
E. Measurement of Kidney ATP Levels 34
F. Measurement of Lipid Hydroperoxide (LPO) and Malondialdehyde (MDA) Levels in the Kidney 34
G. Western blot analysis 34
H. Quantitative Real-time PCR (RT-PCR) 36
I. Statistical analysis 37
Ⅳ. RESULTS 38
A. Effects of echinochrome A on basic parameters in different groups. 38
B. EchA treatment attenuated hypertrophy and fibrosis in the kidney of db/db mice. 40
C. EchA alleviated renal oxidative stress in db/db mice. 44
D. EchA activated AMPK phosphorylation and increases ATP production in diabetic kidneys. 46
E. EchA activated the NRF2/HO-1 pathway contributed to improve mitochondrial function in diabetic kidneys. 48
F. Effect of EchA treatment on PKC/p38MAPK activation and both p53 and c-Jun phosphorylation in the kidney of diabetic db/db mice. 50
Ⅴ. DISCUSSION 55
Ⅵ. CONCLUSION 61
Ⅶ. REFERENCES 63
Table 1. The primers were used in RT-PCR for transcript quantification. 37
Figure 1. The IDF Diabetes Atlas, 2021 21
Figure 2. Pathophysiology of diabetic nephropathy. 23
Figure 3. Protein kinase C (PKC) in diabetic nephropathy 25
Figure 4. The chemical structure, molecular targets, biological functions, and target diseases of Echinochrome A 26
Figure 5. Computer simulated EchA-PKCι docking model 28
Figure 6. Experimental design 31
Figure 7. The effect of EchA treatment on body weight and basic parameters in different groups. 39
Figure 8. The effects of EchA treatment on diabetic renal hypertrophy and fibrosis. 43
Figure 9. The effects of EchA treatment on oxidative stress of db/db mice in the kidney. 45
Figure 10. EchA regulated high glucose-induced p-AMPK and activates ATP production in the kidney of db/db mice. 47
Figure 11. EchA regulated high glucose-induced PGC1-α, NRF2/HO-1 expression in diabetic kidneys. 49
Figure 12. Effect of EchA treatment on PKC family expression in diabetic mice. 52
Figure 13. Effect of EchA treatment on PKCι expression in the renal tissues of db/db mice. 53
Figure 14. Effect of EchA treatment on the phosphorylation p53 as well as c-Jun phosphorylation in the renal tissues of db/db mice. 54
Figure 15. Schematic diagram demonstrating the potential renal-protective mechanism of EchA in DN. 62