Lee Shih-Chieh, Chen Chun-Hao, Yu Chih-Wen, Chen Hsiao Ling, Huang Wei-Tung, Chang Yun-Shiang, Hung Shu-Hsien, Lee Tai-Lin
Department of BioIndustry Technology, Da-Yeh University, Changhua 51591, Taiwan.
Department of Molecular Biotechnology, Da-Yeh University, No. 168, University Rd., Dacun, Changhua 51591, Taiwan.
J Biosci Bioeng. 2016 Sep;122(3):263-9. doi: 10.1016/j.jbiosc.2016.03.002. Epub 2016 Apr 12.
Melanin contributes to skin color, and tyrosinase is the enzyme that catalyzes the initial steps of melanin formation. Therefore, tyrosinase inhibitors may contribute to the control of skin hyperpigmentation. The inhibition of tyrosinase activity by Cinnamomum zeylanicum extracts was previously reported. In this report, we test the hypothesis that Cinnamomum osmophloeum Kanehira, an endemic plant to Taiwan, contains compounds that inhibit tyrosinase activity, similar to C. zeylanicum. The cytotoxicity of three sources of C. osmophloeum Kanehira ethanol extracts was measured in B16-F10 cells using a methyl thiazolyl tetrazolium bromide (MTT) assay. At concentrations greater than 21.25 μg/mL, the ethanol extracts were toxic to the cells; therefore, 21.25 μg/mL was selected to test the tyrosinase activities. At this concentration, all three ethanol extracts decreased the melanin content by 50% in IBMX-induced B16-F10 cells. In addition to the melanin content, greater than 20% of the tyrosinase activity was inhibited by these ethanol extracts. The RT-PCR results showed that tyrosinase and transcription factor MITF mRNAs expression were down-regulated. Consistent with the mRNA results, greater than 40% of the human tyrosinase promoter activity was inhibited based on the reporter assay. Furthermore, our results demonstrate that the ethanol extracts protect cells from UV exposure. C. osmophloeum Kanehira neutralized the IBMX-induced increase in melanin content in B16-F10 cells by inhibiting tyrosinase gene expression at the level of transcription. Moreover, the ethanol extracts also partially inhibited UV-induced cell damage and prevented cell death. Taken together, we conclude that C. osmophloeum Kanehira is a potential skin-whitening and protective agent.
黑色素决定肤色,而酪氨酸酶是催化黑色素形成初始步骤的酶。因此,酪氨酸酶抑制剂可能有助于控制皮肤色素沉着过度。此前有报道称,锡兰肉桂提取物可抑制酪氨酸酶活性。在本报告中,我们验证了一个假设,即台湾本土植物兰屿肉桂含有与锡兰肉桂类似的、能抑制酪氨酸酶活性的化合物。使用甲基噻唑基四氮唑溴盐(MTT)法在B16-F10细胞中检测了三种兰屿肉桂乙醇提取物的细胞毒性。当浓度高于21.25μg/mL时,乙醇提取物对细胞有毒性;因此,选择21.25μg/mL来检测酪氨酸酶活性。在此浓度下,所有三种乙醇提取物均使IBMX诱导的B16-F10细胞中的黑色素含量降低了50%。除黑色素含量外,这些乙醇提取物还抑制了超过20%的酪氨酸酶活性。RT-PCR结果显示,酪氨酸酶和转录因子MITF的mRNA表达下调。与mRNA结果一致,基于报告基因检测,超过40%的人酪氨酸酶启动子活性受到抑制。此外,我们的结果表明,乙醇提取物可保护细胞免受紫外线照射。兰屿肉桂通过在转录水平抑制酪氨酸酶基因表达,中和了IBMX诱导的B16-F10细胞中黑色素含量的增加。此外,乙醇提取物还部分抑制了紫外线诱导的细胞损伤并防止细胞死亡。综上所述,我们得出结论,兰屿肉桂是一种潜在的美白和皮肤保护剂。