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Aminoglycoside 계열 항생제 tobramycin의 멜라닌 생성 증진 효과

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Alternative Title
Effect of Aminoglycoside Antibiotic tobramycin on Melanogenesis Enhancement
Abstract
Since the first antibiotic, penicillin, the era of antibiotics has opened and many new antibiotics have been discovered and developed. However, in the 21st century, antibiotics discovered in the 20th century have been gradually reduced in usage or are no longer used due to antibiotic resistance, side effects, and the development of more effective antibiotics. In this study, we investigated the effects of streptomycin, kanamycin A, kanamycin B, tobramycin, fradiomycin, paromomycin, spertinomycin, hygromycin B, gentamicin, fosfomycin and lincomycin on melanogenesis in B16F10 melanoma cells. First, the effect of each antibiotic on the survival of B16F10 cells was confirmed by MTT assay. Subsequently, the groups of antibiotic alone treatment and the groups treated with antibiotics and α-MSH in the non-toxic concentration were divided to evaluate the effect on melanogenesis. Melanin production was decreased in kanamycin A and streptomycin, and melanin production was increased in tobramycin, fosfomycin and lincomycin. Melanin production was not changed in kanamycin B, fadomycin, paromomycin, spectinomycin, hygromycin B and gentamicin. Among them, melanin contents assay and tyrosinase activity assay were further performed with tobramycin, which increased melanin production remarkably. It was confirmed that tobramycin increased melanin production and increased the activity of tyrosinase, an enzyme important for melanin synthesis. We also confirmed that tobramycin phosphorylates p38, one of the MAPK proteins, by using western blotting and inhibitors, thereby increasing the expression of MITF protein. The increased expression of MITF increased the expressions of tyrosinase, TRP-1, TRP-2, an enzyme important for the synthesis of melanin, in a dose-dependent manner. We compared the effects of kanamycin A, kanamycin B with similar structure to tobramycin and 2-DOS, the central structure of these on the synthesis of melanin. As a result, the effect of tobramycin on the synthesis of melanin was presumed to be the structure of the aminosugar attached to the C-4 of 2-DOS, the central skeleton. Since melanocytes are mainly in the skin layer, the effect of tobramycin on keratinocytes was evaluated. At a concentration of 2 mg/mL, cell viability was over 90 %. In addition, the effect of RAW 264.7 cells on NO production, an inflammatory substance, was evaluated but not effective. Based on these results, we suggest that tobramycin may be helpful in the treatment of hypopigmentation.
Author(s)
文勝鉉
Issued Date
2019
Awarded Date
2019. 8
Type
Dissertation
URI
http://dcoll.jejunu.ac.kr/common/orgView/000000009083
Alternative Author(s)
Moon, Seung Hyun
Affiliation
제주대학교 대학원
Department
대학원 화학과
Advisor
Hyun, Chang Gu
Table Of Contents
ABSTRACT 1
목 차 3
LIST OF TABLE 4
LIST OF FIGURES 5
I. 서 론 7
II. 재료 및 방법 24
1. 시료 및 시약 24
2. 실험방법 26
2.1. 세포배양 26
2.2. 세포독성평가 27
2.3. 멜라닌 함량 측정 28
2.4. Tyrosinase 활성 측정 28
2.5. Western blot 실험 29
2.6. NO 생성량 측정 31
2.7. 통계처리 32
III. 결 과 33
1. B16F10 melanoma 세포 내에서 항생제의 세포독성 평가 33
2. B16F10 melanoma 세포에서 멜라닌 합성에 항생제가 미치는 영향 평가 38
3. Tobramycin이 멜라닌 함량과 tyrosiase 활성에 미치는 영향 50
4. Western blot 분석 53
4.1. 멜라닌 생성 관련 효소들의 발현에 tobramycin이 미치는 영향 53
4.2. MAPK , AKT 신호전달경로에 대한 tobramycin의 영향 56
4.3. 신호전달경로에 대한 저해제 및 tobramycin의 영향 58
5. Aminoglycoside 계열 항생제의 골격 61
6. HaCaT keratinocyte 세포에서 tobramycin의 세포독성 평가 66
7. RAW 264.7 macrophage 세포에서 tobramycin의 NO 활성 억제 효과 68
IV. 고 찰 70
V. 참고문헌 74
Degree
Master
Publisher
제주대학교 대학원
Citation
文勝鉉. (2019). Aminoglycoside 계열 항생제 tobramycin의 멜라닌 생성 증진 효과
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General Graduate School > Chemistry
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