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B16F10 마우스 세포에서 lincomycin의 멜라닌 합성 증진 규명

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Alternative Title
B16F10 mouse cell to identify the enhancement of melanin synthesis of Lincomycin
Abstract
Lincomycin hydrochloride monohydrate (L.H.M), isolated from Streptomyces lincolnensis, is a lincosamide-based antibiotic in the form of a hydrochlorid base. L.H.M was usually used to treat staphylococcus, streptococcus and Bacteroides fragilis infections. Although L.H.M has been utilized in clinical treatment for a long time, it has exhibited several side effects, leading to the introduction of more effective antibiotics, such as clindamycin. Therefore, we conducted an experiment focusing on new possibilities for clinical use of L.H.M. How L.H.M roles in skin melanin production has not been investigated. In this study we used the B16F10 melanoma cell to identify the melanin inducting properties of L.H.M. Melanin contents and tyrosinase activity in the cells were increased by L.H.M without any cytotoxicitiy. Western blot analysis indicated that the protein level of tyrosinase, tyrosinase-related-protenin-1 (TRP-1), and tyrosinase-related-protenin-2 (TRP-2) were increased after L.H.M treatment. In addition, L.H.M enhanced the microphthalmia-associated transmission factor (MITF) expression. Moreover, L.H.M increased phosphorylation of c-Jun N-terminal kinases (JNKs) and p38 mitogen-activated protein kinases in MAPK. Furthermore, the inhibition of tyrosinase activity by L.H.M was stimulated by treatment with SP600125 (a specific JNK inhibitor) and SB203580 (p38 inhibitor). We also found that L.H.M-induced melanogenesis was reversed by H89, which is a specific protein kinase an inhibitor. Finally, using MTT assay, the cell viability of L.H.M in HaCaT cell was confirmed. These results suggested that L.H.M may be used to treat or prevent hypopigmentation disorders or hair depigmentation.
Author(s)
이민숙
Issued Date
2021
Awarded Date
2021. 2
Type
Dissertation
URI
https://oak.jejunu.ac.kr/handle/2020.oak/23479
Alternative Author(s)
Lee, Min Suk
Affiliation
제주대학교 대학원
Department
대학원 화학과
Advisor
현창구
Table Of Contents
ABSTRACT 1
목 차 2
LIST OF FIGURES 4
I. 서론. 6
II. 재료 및 방법 18
1. 시료 및 시약. 18
2. 실험 방법 20
2.1 세포배양 20
2.2 세포독성평가 21
2.3 멜라닌함량측정. 22
2.4 Tyrosinase 활성 측정. 22
2.5 Western blot 실험 23
2.6 통계처리 24
III. 결과. 26
1. B16F10 melanoma 세포 내에서 항생제들의 세포독성 평가 26
2. B16F10 melanoma 세포 내에서 멜라닌 함성에 항생제들이 미치는 영향 31
3. B16F10 melanoma 세포 내에서 LHM의 Tyrosinase 활성에 미치는 영향 36
4. Western blot 분석 38
4.1 멜라닌 생성 관련 효소들의 발현에 LHM이 미치는 영향 38
4.2 MAPK 신호전달경로에 대한 LHM의 영향 41
4.3 신호전달경로에 대한 저헤제 및 LHM의 영향 43
4.4 B16F10 세포 내에서 LHM의 Akt 인산화 저해. 45
4.5 PKA 의존 신호 경로를 통한 LHM의 멜라닌 생성 유도 47
5. HaCaT keratinocyte 세포에서 LHM의 독성 평가 49
IV. 고찰. 51
V. 참고문헌. 55
Degree
Master
Publisher
제주대학교 대학원
Citation
이민숙. (2021). B16F10 마우스 세포에서 lincomycin의 멜라닌 합성 증진 규명
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General Graduate School > Chemistry
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