遗传 ›› 2017, Vol. 39 ›› Issue (12): 1178-1187.doi: 10.16288/j.yczz.17-178

• 研究报告 • 上一篇    

金线莲抑制斑马鱼黑色素形成的活性组分筛选及机理研究

许璟瑾1(),张文娟1,王静怡1,姚丽云1,潘裕添1,欧一新1,2,薛钰1,3()   

  1. 1. 闽南师范大学,福建省菌类活性物质工程技术研究中心,漳州 363000
    2. 上海交通大学生命科学技术学院,上海 200240
    3. 膜生物学国家重点实验室,北京 100101
  • 收稿日期:2017-05-15 修回日期:2017-08-02 出版日期:2017-12-20 发布日期:2017-12-20
  • 作者简介:许璟瑾,在读硕士研究生,专业方向：化学生物学。E-mail: 13123352787@163.com|薛钰，博士，讲师，研究方向：细胞与发育生物学。E-mail: xueyu0614@163.com
  • 基金资助:
    福建省自然科学基金项目(2015J05070);福建省教育厅科技项目(JK2015028);闽南师范大学杰出青年科研人才计划(MJ14007)

The active component screening of Anoectochilus roxburghii and the functional study on inhibition of melanogenesis in zebrafish

Jingjin Xu1(),Wenjuan Zhang1,Jingyi Wang1,Liyun Yao1,Yutian Pan1,Yixin Ou1,2,Yu Xue1,3()   

  1. 1. Fujian Engineering Technology Research Center of Mushroom Industry, Minnan Normal University, Zhangzhou 363000, China
    2. School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China
    3. State Key Laboratory of Membrane Biology, Beijing 100101, China
  • Received:2017-05-15 Revised:2017-08-02 Published:2017-12-20 Online:2017-12-20
  • Supported by:
    the Natural Science Foundation of Fujian Province(2015J05070);the Education Scientific Research Project of Fujian Province(JK2015028);Outstanding Youth Research Personnel Training Program of MinnanNormal University(MJ14007)

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摘要：

为探索金线莲中对黑色素形成具有抑制效果的活性组分,本研究对金线莲进行分离、提取,获得总提组、醇沉组与醇提组,利用斑马鱼筛选金线莲具有美白作用的活性组分。将受精后0.75 h的斑马鱼胚胎分别暴露于不同浓度的金线莲总提组、醇沉组、醇提组,72 h时观察结果表明,金线莲醇提物能有效抑制斑马鱼胚胎黑色素和黄色素沉着,浓度越高抑制效果越明显,且不影响胚胎生长发育。进一步采用半定量PCR和整胚原位杂交技术定量和定性地检测黑色素形成相关基因mRNA表达,结果表明金线莲醇提物可以有效降低silv、tyr和tyrp1a等黑色素合成相关基因的转录水平,且具有浓度依赖关系。通过检测酪氨酸酶活性显示,加入醇提物的实验组其酪氨酸酶活性随着处理浓度升高而逐渐降低。此外,在黑色素已经大量形成的情况下,金线莲醇提物仍可通过下调黑色素合成相关基因的mRNA表达及酪氨酸酶活性来抑制黑色素的形成,并且这种抑制效果可在金线莲醇提物撤除后得到恢复。上述实验结果表明,金线莲醇提物能显著抑制斑马鱼黑色素的形成,本文为金线莲在美白产品领域的开发和应用方面提供了有力支撑。

关键词: 斑马鱼, 金线莲醇提物, 黑色素, 基因表达, 酪氨酸酶

Abstract:

The aim of this study is to explore the active components of Anoectochilus roxburghii capable of inhibiting melanin formation using chemical separation and extraction and functional analysis. Anoectochilus roxburghii were extracted with alcohol and separated into three groups: the total extraction group, alcohol extracted group and alcohol precipitated group. Zebrafish embryos at 0.75 h post-fertilization were exposed to various concentrations of the three groups of extracts, and analyzed at 72 h, using semi-quantitative RT-PCR and in situ hybridization. The results showed that the alcohol extracts inhibit melanogenesis most significantly in the zebrafish embryos. The mRNAs of melanin-related genes, such as silv, tyr, tyrp1a, were down-regulated by the alcohol extracts spatially and temporally. The alcohol extracts also inhibited the activity of tyrosinase, a key enzyme in melanogenesis, in a dosage dependent manner. In addition, the alcohol extracts also display a remarkable inhibitory effect on melanin synthesis through down-regulation of mRNAs of melanin-related genes and tyrosinase activity in zebrafish embryos, in which a large amount of melanin has already been synthesized. Such inhibitory effect could be reversed after the withdrawal of the alcohol extracts. Our results showed that the alcohol extracts of Anoectochilus roxburghii can significantly inhibit zebrafish melanogenesis, supporting the notion that Anoectochilus roxburghii could potentially be used in the development and production of natural whitening products.

Key words: zebrafish, Anoectochilus roxburghii alcohol extracts, melanin, gene expression, tyrosinase