Growing consumer interest in skin whitening has led to intensive investigations of whitening methods. In this study, we evaluated the effect of sphingomyelin, a component of cell membranes, on melanin production. B16 mouse melanoma cells were treated with lauroyl-sphingomyelin (SM) or its metabolite lauroyl-ceramide (CER) and measured for cell viability, melanin content, and direct and indirect tyrosinase activity. Expression of melanin synthesis-related genes encoding tyrosinase (Tyr), tyrosinase-related proteins (Trp1 and Trp2), and microphthalmia-associated transcription factor (Mitf) were quantified by real-time PCR, and SM content in cells was measured by fluorescence high-performance liquid chromatography. SM treatment decreased melanin content in a concentration-dependent manner, without significantly altering the number of viable cells. By contrast, treatment with CER at the same concentrations did not decrease melanin content. SM inhibited the activity of intracellular tyrosinase, but not mushroom-derived tyrosinase. Gene expression levels of Tyr and Mitf were significantly reduced by treatment with SM, while those of Trp2 and Mitf were significantly reduced by CER. Fluorescence-labeled SM was converted to fluorescence-labeled CER in cells over time. In conclusion, CER was found to inhibit melanogenesis without inhibiting tyrosinase activity, suggesting that SM is more water soluble than CER, and is more effectively taken up into cells.
Keywords: Ceramide; Inhibition; Melanin; Real-time PCR; Sphingomyelin; Tyrosinase.
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