Title Page
Contents
ABSTRACT 9
1. INTRODUCTION 11
2. MATERIALS AND METHODS 20
2-1. Materials 20
2-2. Cell culture 20
2-3. Cell viability assay 21
2-4. Tyrosinase zymography 21
2-5. Melanin content measurement 22
2-6. Western blot analysis 22
2-7. Immunocytochemistry 23
2-8. siRNA-mediated knockdown 24
3. RESULTS AND DISCUSSION 25
3-1. Effects of BBC on cytotoxicity in murine melanoma cell line (B16-F1) 25
3-2. Inhibitory effects of BBC on intracellular tyrosinase activity 25
3-3. Effects of BBC on ERK pathway and various melanogenesis-associated proteins in B16-F1 cells 28
3-4. Inhibitory effects of BBC on melanin synthesis in B16-F1 cells 28
3-5. Effects of BBC on MAPK pathway in B16-F1 cells 31
3-6. Effect of BBC on autophagy system in B16-F1 cells 31
3-7. Immunocytochemistry analysis of the induction of autophagy by BBC in B16-F1 cells 33
3-8. Effects of BBC on the production of melanin synthesis in B16-F1 cells transfected with siRNAs specific to autophagy-related genes 33
4. Conclusion 36
5. 적요 40
6. REFERENCES 42
Fig. 1. Schematic diagram showing the structure of young and aged skins. 12
Fig. 2. Synthetic pathway of melanin pigments. 13
Fig. 3. Signaling pathway of melanogenesis in melanocytes. 14
Fig. 4. Biochemical pathway of melanin synthesis. 15
Fig. 5. Signaling pathway of autophagy. 17
Fig. 6. Chemical structure of berberine chloride (BBC). 18
Fig. 7. Effects of BBC on cytotoxicity in murine melanoma cell line (B16-F1). 26
Fig. 8. Inhibitory effects of BBC on intracellular tyrosinase activity. 27
Fig. 9. Effects of BBC on ERK pathway and various melanogenesis-associated proteins in B16-F1 cells. 29
Fig. 10. Inhibitory effects of BBC on melanin synthesis in B16-F1 cells. 30
Fig. 11. Effects of BBC on MAPK pathway in B16-F1 cells. 32
Fig. 12. Effect of BBC on autophagy system in B16-F1 cells. 34
Fig. 13. Immunocytochemistry analysis of the induction of autophagy by BBC in B16-F1 cells. 35
Fig. 14. Effects of BBC on the production of melanin synthesis in B16-F1 cells transfected with siRNAs specific to autophagy-related genes. 37
Fig. 15. Summary of possible inhibitory effects of BBC on melanogenesis. 39