표제지
목차
ABSTRACT 10
제1장 서론 11
제2장 실험 및 방법 14
1. 시약 14
2. 세포 배양 14
3. 세포 생존율 측정 15
4. 세포 내 멜라닌 함량 측정 16
5. Cellular tyrosinase 활성 측정 17
6. Mushroom tyrosinase 활성 측정 17
7. Immunoblotting analysis 18
8. 멜라닌 합성 관련 mRNA 발현 분석 20
9. 통계 분석 21
제3장 연구 결과 22
1. 토양 유래 멜라닌 합성 저해 효능 방선균 추출물 스크리닝 22
2. B16F10 흑색종 세포 내 Embleya sp. AN120556 유래 추출물에 의한 세포 생존율 평가 24
3. B16F10 흑색종 세포 내 α-MSH 유도 멜라닌 생성에 대한 Embleya sp. AN120556 유래 추출물의 억제 효능 26
4. B16F10 흑색종 세포 내 ESB에 의한 Cellular Tyrosinase 활성 분석 29
5. B16F10 흑색종 세포에서 ESB 추출 용매 별 멜라닌 합성 저해 효능 분석 31
6. B16F10 흑색종 세포에서 ESBH에 의한 멜라닌 합성 관련 작용 효소의 단백질 및 유전자 발현 분석 39
7. B16F10 흑색종 세포에서 ESBH에 의한 Mitf 단백질 및 유전자 발현 분석 42
8. B16F10 흑색종 세포에서 ESBH에 의한 멜라닌 생합성 저해 기전 규명 45
9. B16F10 흑색종 세포에서 Embleya 속 균주 추출물 별 멜라닌 합성 저해 효능 분석 49
제4장 고찰 53
제5장 결론 58
참고문헌 59
국문초록 66
Table 2-1. List of antibodies used for western blot analysis. 19
Table 2-2. List of primer sequences for qRT-PCR analysis 20
Table 3-1. Screening of skin whitening microbes from actinomycetes library 23
Figure 3-1. Effect of Embleya sp. AN120556-derived extracts on cell viability in B16F10 cells. 25
Figure 3-2. Effect of Embleya sp. AN120556-derived extracts on melanin contents in α-MSH stimulated B16F10 cells. 27
Figure 3-3. Effect of ESB on melanin contents in α-MSH stimulated B16F10 cells. 28
Figure 3-4. Effect of ESB on cellular tyrosinase activity in α-MSH stimulated B16F10 cells. 30
Figure 3-5. Embleya sp. AN120556-derived fractions extracted with 5 different solvents. 33
Figure 3-6. Effect of ESB extracted with 5 different solvents on cell viability in B16F10 cells. 34
Figure 3-7. Effect of ESB extracted with 5 different solvents on melanogenesis in α-MSH stimulated B16F10 cells. 35
Figure 3-8. Effect of ESBH on melanogenesis in α-MSH stimulated B16F10 cells. 36
Figure 3-9. Effect of ESBH on cellular tyrosinase activity in α-MSH stimulated B16F10 cells 37
Figure 3-10. Effect of ESBH on mushroom tyrosinase activity. 38
Figure 3-11. Effect of ESBH on the expression of melanogenesis-related proteins in α-MSH stimulated B16F10 cells. 40
Figure 3-12. Effect of ESBH on mRNA expression of melanogenesis-related proteins in α-MSH stimulated B16F10 cells. 41
Figure 3-13. Effects of ESBH on the protein expression of Mitf in α-MSH stimulated B16F10 cells. 43
Figure 3-14. Effects of ESBH on mRNA expression of Mitf in α-MSH stimulated B16F10 cells. 44
Figure 3-15. Effect of ESBH on phosphorylation level of Pka and Creb in α-MSH stimulated B16F10 cells. 46
Figure 3-16. Effect of ESBH on phosphorylation level of Mapk in α-MSH stimulated B16F10 cells. 47
Figure 3-17. Effect of ESBH on phosphorylation level of Msk1 in α-MSH stimulated B16F10 cells. 48
Figure 3-18. Effect of Embleya sp. NBRC100760 extract on cell viability in B16F10 cells. 50
Figure 3-19. Effect of Embleya sp. NBRC13850 extract on cell viability in B16F10 cells. 51
Figure 3-20. Effect of Embleya sp. microbes extracts on melanin contents in α-MSH stimulated B16F10 cells. 52