B16F10 마우스 세포에서 lincomycin의 멜라닌 합성 증진 규명

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.