Abstract
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.
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The data presented in this study are available on request from the corresponding author.
References
Briganti S, Camera E, Picardo M (2003) Chemical and instrumental approaches to treat hyperpigmentation. Pigment Cell Res 16:101–110. https://doi.org/10.1034/j.1600-0749.2003.00029.x
Burnside B, Evans M, Fletcher RT, Chader GJ (1982) Induction of dark-adaptive retinomotor movement (cell elongation) in teleost retinal cones by cyclic adenosine 3ʹʹ,5-monophosphate. J Gen Physiol 79:759–774. https://doi.org/10.1085/jgp.79.5.759
Chedekel MR, Murr BL, Zeise L (1992) Melanin standard method: empirical formula. Pigment Cell Res 5:143–147. https://doi.org/10.1111/j.1600-0749.1992.tb00010.x
D’Mello SA, Finlay GJ, Baguley BC, Askarian-Amiri ME (2016) Signaling pathways in melanogenesis. Int J Mol Sci 17:1144. https://doi.org/10.3390/ijms17071144
Hait NC, Oskeritzian CA, Paugh SW, Milstien S, Spiegel S (2006) Sphingosine kinases, sphingosine 1-phosphate, apoptosis and diseases. Biochim Biophys Acta 1758:2016–2026. https://doi.org/10.1016/j.bbamem.2006.08.007
Hannun YA (1994) The sphingomyelin cycle and the second messenger function of ceramide. J Biol Chem 269:3125–3128. https://doi.org/10.1016/S0021-9258(17)41834-5
Kameyama K, Takemura T, Hamada Y, Sakai C, Kondoh S, Nishiyama S, Urabe K, Hearing VJ (1993) Pigment production in murine melanoma cells is regulated by tyrosinase, tyrosinase-related protein 1 (TRP1), DOPAchrome tautomerase (TRP2), and a melanogenic inhibitor. J Invest Dermatol 100:126–131. https://doi.org/10.1111/1523-1747.ep12462778
Kim DS, Kim SY, Chung JH, Kim KH, Eun HC, Park KC (2002) Delayed ERK activation by ceramide reduces melanin synthesis in human melanocytes. Cell Signal 14:779–785. https://doi.org/10.1016/s0898-6568(02)00024-4
Kim DS, Hwang ES, Lee JE, Kim SY, Kwon SB, Park KC (2003) Sphingosine-1-phosphate decreases melanin synthesis via sustained ERK activation and subsequent MITF degradation. J Cell Sci 116:1699–1706. https://doi.org/10.1242/jcs.00366
Kim DS, Jeong YM, Park IK, Hahn HG, Lee HK, Kwon SB, Jeong JH, Yang SJ, Sohn UD, Park KC (2007) A new 2-imino-1,3-thiazoline derivative, KHG22394, inhibits melanin synthesis in mouse B16 melanoma cells. Biol Pharm Bull 30:180–183. https://doi.org/10.1248/bpb.30.180
Kim DS, Park SH, Kwon SB, Kwon NS, Park KC (2010) Sphingosylphosphorylcholine inhibits melanin synthesis via pertussis toxin-sensitive MITF degradation. J Pharm Pharmacol 62:181–187. https://doi.org/10.1211/jpp.62.02.0005
Kobayashi T, Urabe K, Winder A, Jiménez-Cervantes C, Imokawa G, Brewington T, Solano F, García-Borrón JC, Hearing VJ (1994) Tyrosinase related protein 1 (TRP1) functions as a DHICA oxidase in melanin biosynthesis. EMBO J 13:5818–5825. https://doi.org/10.1002/j.1460-2075.1994.tb06925.x
Leung YH, Bäßler SC, Koch C, Scheu T, Meyer U, Dänicke S, Huber K, Kenéz Á (2020) Sphingolipid profiling reveals different extent of ceramide accumulation in bovine retroperitoneal and subcutaneous adipose tissues. Metabolites 10:473. https://doi.org/10.3390/metabo10110473
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Mann T, Gerwat W, Batzer J, Eggers K, Scherner C, Wenck H, Stäb F, Hearing VJ, Röhm KH, Kolbe L (2018) Inhibition of human tyrosinase requires molecular motifs distinctively different from mushroom tyrosinase. J Invest Dermatol 138:1601–1608. https://doi.org/10.1016/j.jid.2018.01.019
Mao C, Obeid LM (2008) Ceramidases: regulators of cellular responses mediated by ceramide, sphingosine, and sphingosine-1-phosphate. Biochim Biophys Acta 1781:424–434. https://doi.org/10.1016/j.bbalip.2008.06.002
Nomoto K, Itaya Y, Watanabe K, Yamashita T, Okazaki T, Tokudome Y (2018) Epidermal permeability barrier function and sphingolipid content in the skin of sphingomyelin synthase 2 deficient mice. Exp Dermatol 27:827–832. https://doi.org/10.1111/exd.13497
Palumbo A, d’Ischia M, Misuraca G, Prota G (1991) Mechanism of inhibition of melanogenesis by hydroquinone. Biochim Biophys Acta 1073:85–90. https://doi.org/10.1016/0304-4165(91)90186-k
Parvez S, Kang M, Chung HS, Cho C, Hong MC, Shin MK, Bae H (2006) Survey and mechanism of skin depigmenting and lightening agents. Phytother Res 20:921–934. https://doi.org/10.1002/ptr.1954
Videira IF, Moura DF, Magina S (2013) Mechanisms regulating melanogenesis. An Bras Dermatol 88:76–83. https://doi.org/10.1590/s0365-05962013000100009
Wakamatsu K, Kavanagh R, Kadekaro AL, Terzieva S, Sturm RA, Leachman S, Abdel-Malek Z, Ito S (2006) Diversity of pigmentation in cultured human melanocytes is due to differences in the type as well as quantity of melanin. Pigment Cell Res 19:154–162. https://doi.org/10.1111/j.1600-0749.2006.00293.x
Zhang KQ, Lin LL, Xu HJ (2022) Research on antioxidant performance of diglucosyl gallic acid and its application in emulsion cosmetics. Int J Cosmet Sci 44:177–188. https://doi.org/10.1111/ics.12766
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We thank Michelle Kahmeyer-Gabbe, PhD, from Edanz Group (https://en-author-services.edanzgroup.com/) for editing a draft of this manuscript.
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YT: Study conceptualization and design. MF: Acquisition, analysis, and interpretation of data. YT and MF: Drafting and critically revising the manuscript for important intellectual content. YT and MF: Final approval of the version to be published. Both authors read and accepted the manuscript.
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Tokudome, Y., Fukutomi, M. Sphingomyelin reduces melanogenesis in murine B16 melanoma cells through indirect suppression of tyrosinase. Cytotechnology 75, 93–101 (2023). https://doi.org/10.1007/s10616-022-00562-y
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DOI: https://doi.org/10.1007/s10616-022-00562-y