{"title":"普托瑞辛通过调节 B16F1 小鼠黑色素瘤细胞中的 MITF 转录因子上调黑色素生成","authors":"Natchanok Talapphet, Moon-Moo Kim","doi":"10.17113/ftb.62.01.24.8120","DOIUrl":null,"url":null,"abstract":"Research background. Aging is a biochemical, metabolic, and genetic physiological phenomenon. The suppression of melanin biosynthesis, evident in the graying of hair, is a hallmark of aging resulting from translation failure, reduced enzyme activity, and cellular senescence. Putrescine, the smallest member of the polyamine family and an organic chemical, is present in living mammalian cells, playing a crucial role in regulating skin melanogenesis. Therefore, the purpose of this study is to explore the effect of putrescine on the signaling pathways of melanogenesis in melanoma cells.\nExperimental approach. Putrescine was studied on the melanin production capacity was examined through a tyrosinase activity assay. To assess the cell viability of B16F1 cells exposed to putrescine, an MTT assay was performed. The impact of putrescine on melanin synthesis in the presence of H2O2 was evaluated using various in vitro assays in B16F1 cells. The effect of putrescine on melanin production in B16F1 cells was achieved through a dedicated melanin production assay. Gene expression analysis was conducted using RT-PCR. Furthermore, the impact of putrescine on the expression of proteins related to melanin production in H2O2-treated cells was examined through immunofluorescence and western blot analysis. \nResults and conclusions. Putrescine increased tyrosinase activity and demonstrated non-cytotoxicity in B16F1 cells. Furthermore, putrescine effectively scavenged H2O2, as evidenced by the reduction in intracellular H2O2 levels in DCFH-DA analysis, and promoted melanin production in living cells. The stimulation of melanogenesis by putrescine was attributed to the elevated expression of Mitf, Tyr, Trp-1, and Trp-2 genes. Immunofluorescence investigations revealed that putrescine enhanced the expression of proteins associated with melanogenesis and upregulated TYR, TRP-1, and TRP-2 via the MITF transcription factor and increased the expression of MSRA and MSRB in H2O2-treated cells, thereby effectively promoting melanogenesis. These findings suggest that putrescine may be utilized to stimulate melanin synthesis.\nNovelty and scientific contribution. Putrescine could be solely used as a cosmetic agent to prevent premature graying of hair.","PeriodicalId":12400,"journal":{"name":"Food Technology and Biotechnology","volume":"137 40","pages":""},"PeriodicalIF":2.3000,"publicationDate":"2024-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Putrescine Upregulates Melanogenesis Through Modulation of MITF Transcription Factor in B16F1 Mouse Melanoma Cells\",\"authors\":\"Natchanok Talapphet, Moon-Moo Kim\",\"doi\":\"10.17113/ftb.62.01.24.8120\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Research background. Aging is a biochemical, metabolic, and genetic physiological phenomenon. The suppression of melanin biosynthesis, evident in the graying of hair, is a hallmark of aging resulting from translation failure, reduced enzyme activity, and cellular senescence. Putrescine, the smallest member of the polyamine family and an organic chemical, is present in living mammalian cells, playing a crucial role in regulating skin melanogenesis. Therefore, the purpose of this study is to explore the effect of putrescine on the signaling pathways of melanogenesis in melanoma cells.\\nExperimental approach. Putrescine was studied on the melanin production capacity was examined through a tyrosinase activity assay. To assess the cell viability of B16F1 cells exposed to putrescine, an MTT assay was performed. The impact of putrescine on melanin synthesis in the presence of H2O2 was evaluated using various in vitro assays in B16F1 cells. The effect of putrescine on melanin production in B16F1 cells was achieved through a dedicated melanin production assay. Gene expression analysis was conducted using RT-PCR. Furthermore, the impact of putrescine on the expression of proteins related to melanin production in H2O2-treated cells was examined through immunofluorescence and western blot analysis. \\nResults and conclusions. Putrescine increased tyrosinase activity and demonstrated non-cytotoxicity in B16F1 cells. Furthermore, putrescine effectively scavenged H2O2, as evidenced by the reduction in intracellular H2O2 levels in DCFH-DA analysis, and promoted melanin production in living cells. The stimulation of melanogenesis by putrescine was attributed to the elevated expression of Mitf, Tyr, Trp-1, and Trp-2 genes. Immunofluorescence investigations revealed that putrescine enhanced the expression of proteins associated with melanogenesis and upregulated TYR, TRP-1, and TRP-2 via the MITF transcription factor and increased the expression of MSRA and MSRB in H2O2-treated cells, thereby effectively promoting melanogenesis. These findings suggest that putrescine may be utilized to stimulate melanin synthesis.\\nNovelty and scientific contribution. Putrescine could be solely used as a cosmetic agent to prevent premature graying of hair.\",\"PeriodicalId\":12400,\"journal\":{\"name\":\"Food Technology and Biotechnology\",\"volume\":\"137 40\",\"pages\":\"\"},\"PeriodicalIF\":2.3000,\"publicationDate\":\"2024-01-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Food Technology and Biotechnology\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://doi.org/10.17113/ftb.62.01.24.8120\",\"RegionNum\":4,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Food Technology and Biotechnology","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.17113/ftb.62.01.24.8120","RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
Putrescine Upregulates Melanogenesis Through Modulation of MITF Transcription Factor in B16F1 Mouse Melanoma Cells
Research background. Aging is a biochemical, metabolic, and genetic physiological phenomenon. The suppression of melanin biosynthesis, evident in the graying of hair, is a hallmark of aging resulting from translation failure, reduced enzyme activity, and cellular senescence. Putrescine, the smallest member of the polyamine family and an organic chemical, is present in living mammalian cells, playing a crucial role in regulating skin melanogenesis. Therefore, the purpose of this study is to explore the effect of putrescine on the signaling pathways of melanogenesis in melanoma cells.
Experimental approach. Putrescine was studied on the melanin production capacity was examined through a tyrosinase activity assay. To assess the cell viability of B16F1 cells exposed to putrescine, an MTT assay was performed. The impact of putrescine on melanin synthesis in the presence of H2O2 was evaluated using various in vitro assays in B16F1 cells. The effect of putrescine on melanin production in B16F1 cells was achieved through a dedicated melanin production assay. Gene expression analysis was conducted using RT-PCR. Furthermore, the impact of putrescine on the expression of proteins related to melanin production in H2O2-treated cells was examined through immunofluorescence and western blot analysis.
Results and conclusions. Putrescine increased tyrosinase activity and demonstrated non-cytotoxicity in B16F1 cells. Furthermore, putrescine effectively scavenged H2O2, as evidenced by the reduction in intracellular H2O2 levels in DCFH-DA analysis, and promoted melanin production in living cells. The stimulation of melanogenesis by putrescine was attributed to the elevated expression of Mitf, Tyr, Trp-1, and Trp-2 genes. Immunofluorescence investigations revealed that putrescine enhanced the expression of proteins associated with melanogenesis and upregulated TYR, TRP-1, and TRP-2 via the MITF transcription factor and increased the expression of MSRA and MSRB in H2O2-treated cells, thereby effectively promoting melanogenesis. These findings suggest that putrescine may be utilized to stimulate melanin synthesis.
Novelty and scientific contribution. Putrescine could be solely used as a cosmetic agent to prevent premature graying of hair.
期刊介绍:
Food Technology and Biotechnology (FTB) is a diamond open access, peer-reviewed international quarterly scientific journal that publishes papers covering a wide range of topics, including molecular biology, genetic engineering, biochemistry, microbiology, biochemical engineering and biotechnological processing, food science, analysis of food ingredients and final products, food processing and technology, oenology and waste treatment.
The Journal is published by the University of Zagreb, Faculty of Food Technology and Biotechnology, Croatia. It is an official journal of Croatian Society of Biotechnology and Slovenian Microbiological Society, financed by the Croatian Ministry of Science and Education, and supported by the Croatian Academy of Sciences and Arts.