{"title":"鞘磷脂通过间接抑制酪氨酸酶减少小鼠 B16 黑色素瘤细胞的黑色素生成。","authors":"Yoshihiro Tokudome, Moeko Fukutomi","doi":"10.1007/s10616-022-00562-y","DOIUrl":null,"url":null,"abstract":"<p><p>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 (<i>Tyr</i>), tyrosinase-related proteins <i>(Trp1</i> and <i>Trp2</i>), and microphthalmia-associated transcription factor (<i>Mitf</i>) 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 <i>Tyr</i> and <i>Mitf</i> were significantly reduced by treatment with SM, while those of <i>Trp2</i> and <i>Mitf</i> 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.</p>","PeriodicalId":2,"journal":{"name":"ACS Applied Bio Materials","volume":null,"pages":null},"PeriodicalIF":4.6000,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10030692/pdf/","citationCount":"0","resultStr":"{\"title\":\"Sphingomyelin reduces melanogenesis in murine B16 melanoma cells through indirect suppression of tyrosinase.\",\"authors\":\"Yoshihiro Tokudome, Moeko Fukutomi\",\"doi\":\"10.1007/s10616-022-00562-y\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>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 (<i>Tyr</i>), tyrosinase-related proteins <i>(Trp1</i> and <i>Trp2</i>), and microphthalmia-associated transcription factor (<i>Mitf</i>) 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 <i>Tyr</i> and <i>Mitf</i> were significantly reduced by treatment with SM, while those of <i>Trp2</i> and <i>Mitf</i> 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.</p>\",\"PeriodicalId\":2,\"journal\":{\"name\":\"ACS Applied Bio Materials\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":4.6000,\"publicationDate\":\"2023-04-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10030692/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"ACS Applied Bio Materials\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1007/s10616-022-00562-y\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2022/11/30 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q2\",\"JCRName\":\"MATERIALS SCIENCE, BIOMATERIALS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Applied Bio Materials","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1007/s10616-022-00562-y","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2022/11/30 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"MATERIALS SCIENCE, BIOMATERIALS","Score":null,"Total":0}
引用次数: 0
摘要
消费者对皮肤美白的兴趣与日俱增,促使人们对美白方法进行深入研究。在这项研究中,我们评估了细胞膜成分鞘磷脂对黑色素生成的影响。用月桂酰鞘磷脂(SM)或其代谢物月桂酰神经酰胺(CER)处理 B16 小鼠黑色素瘤细胞,并测定细胞活力、黑色素含量以及直接和间接酪氨酸酶活性。通过实时 PCR 对编码酪氨酸酶(Tyr)、酪氨酸酶相关蛋白(Trp1 和 Trp2)和小眼病相关转录因子(Mitf)的黑色素合成相关基因的表达进行了定量,并通过荧光高效液相色谱法测定了细胞中的 SM 含量。SM处理以浓度依赖的方式降低了黑色素含量,但并没有显著改变存活细胞的数量。相比之下,用相同浓度的 CER 处理不会降低黑色素含量。SM能抑制细胞内酪氨酸酶的活性,但不能抑制蘑菇衍生的酪氨酸酶。SM能显著降低Tyr和Mitf的基因表达水平,而CER则能显著降低Trp2和Mitf的基因表达水平。随着时间的推移,荧光标记的 SM 在细胞中转化为荧光标记的 CER。总之,研究发现 CER 可抑制黑色素生成而不抑制酪氨酸酶的活性,这表明 SM 比 CER 更易溶于水,能更有效地被细胞吸收。
Sphingomyelin reduces melanogenesis in murine B16 melanoma cells through indirect suppression of tyrosinase.
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.