水仙醇提物对斑马鱼胚胎黑色素合成的抑制作用

摘要

目的 以斑马鱼为模型,重点研究水仙鳞茎的醇提物对斑马鱼黑色素合成的抑制效应.方法 将斑马鱼胚胎暴露在0、50、100、200、500 μg/mL不同浓度的水仙花醇提物中,观察其黑色素生成情况,并进一步测定体内黑色素合成通路关键蛋白酪氨酸酶的酶活性变化和其他标记基因的时空表达,同时以200 μg/mL的熊果苷(arbutin,Ar)处理组为阳性对照.结果 定性观察表明,随着水仙提取物处理浓度的增加,对斑马鱼黑色素合成的抑制明显增强;进一步检测黑色素合成相关基因的时空表达,证明了水仙醇提物通过抑制酪氨酸酶(TYR)、银色毛发(SILV)和Mitfa等基因的mRNA表达而抑制黑色素合成;最后,通过检测酪氨酸酶的酶活性,发现随着醇提物浓度的增加导致酶活性逐渐降低.结论 水仙醇提物能有效地抑制斑马鱼胚胎黑色素的生成,同时该研究也为斑马鱼模型在天然美白化合物筛选和产品开发的应用提供了研究基础和思路.%Objective The aim of this study was to explore the inhibitory effect of alcohol extracts of Narcissus bulb on melanogenesis in zebrafish embryos.Methods Zebrafish embryos were exposed to different concentrations of alcohol extracts of Narcissus (0, 50, 100, 200, and 500 μg/mL), and then the formation of melanin was observed.Furthermore, we measured the enzyme activity of tyrosinase (TYR), which plays a pivotal role in melanogenesis.Meanwhile, the spatial and temporal pattern of melanin-specific marker genes were detected.Arbutin (Ar) was used as a positive control in all these experiments.Results The treatment of alcohol extracts of Narcissus bulb inhibited melanogenesis in the zebrafish embryos in a dose-dependent manner.Furthermore, the mRNA expression levels of melanin-related genes such as tyrosinase (TYR), silver (SILV) and Mitfa were significantly reduced after treatment with different concentrations of Narcissus extracts by in-situ and semi-quantitative PCR.Finally, the enzyme activity of tyrosinase was also gradually decreased with increasing concentrations of the alcohol extracts.Conclusions Narcissus alcohol extracts can effectively inhibit the production of melanin in zebrafish embryos.This study provides potential evidence and approaches for the screening of natural whitening compounds using zebrafish models.