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Bio-Design and Manufacturing  2021 Vol.4 No.4 P.806-817

http://doi.org/10.1007/s42242-021-00138-3


Evaluation ofaresorufin?based fuorescent probe fortyrosinase detection inskin pigmentation disorders


Author(s):  YiboHu, HongliangZeng, JiaxingJiang, ShengYang, JinhuaHuang, JingChen, QinghaiZeng

Affiliation(s):  Department ofDermatology, The Third Xiangya Hospital ofCentral South University, Tongzipo Road No.138, Yuelu District, Changsha410013, China; more

Corresponding email(s):   43700351@qq.com, zengqinghai@csu.edu.cn

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YiboHu, HongliangZeng, JiaxingJiang, ShengYang, JinhuaHuang, JingChen, QinghaiZeng. Evaluation ofaresorufin?based fuorescent probe fortyrosinase detection inskin pigmentation disorders[J]. Journal of Zhejiang University Science D, 2021, 4(4): 806-817.

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author="YiboHu, HongliangZeng, JiaxingJiang, ShengYang, JinhuaHuang, JingChen, QinghaiZeng",
journal="Journal of Zhejiang University Science D",
volume="4",
number="4",
pages="806-817",
year="2021",
publisher="Zhejiang University Press & Springer",
doi="10.1007/s42242-021-00138-3"
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%0 Journal Article
%T Evaluation ofaresorufin?based fuorescent probe fortyrosinase detection inskin pigmentation disorders
%A YiboHu
%A HongliangZeng
%A JiaxingJiang
%A ShengYang
%A JinhuaHuang
%A JingChen
%A QinghaiZeng
%J Journal of Zhejiang University SCIENCE D
%V 4
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%P 806-817
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%D 2021
%I Zhejiang University Press & Springer
%DOI 10.1007/s42242-021-00138-3

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T1 - Evaluation ofaresorufin?based fuorescent probe fortyrosinase detection inskin pigmentation disorders
A1 - YiboHu
A1 - HongliangZeng
A1 - JiaxingJiang
A1 - ShengYang
A1 - JinhuaHuang
A1 - JingChen
A1 - QinghaiZeng
J0 - Journal of Zhejiang University Science D
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PB - Zhejiang University Press & Springer
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DOI - 10.1007/s42242-021-00138-3


Abstract: 
Purpose Skin pigmentation disorders, such as vitiligo and melasma, are difcult to diagnose in the early stages, but abnormal tyrosinase levels and tyrosinase activity are potential indicators. Some resorufn-based fuorescence probes (RBFPs) have been designed to detect tyrosinase in tumors, but they have not been used in skin pigmentation disorders. In this study, one of these RBFPs (synthesized by resorufn salt coupled with 3-(bromomethyl)phenol) was evaluated comprehensively. Methods The RBFP was tested in diferent kinds of mouse and human skin cells, as well as in invivo models, including zebrafsh, guinea pigs, and SpragueDawley rats. In addition, small interfering RNAs (siRNAs), kojic acid, and 1-phenyl- 2-thiourea (PTU) were used to inhibit tyrosinase levels or tyrosinase activity. Results This probe successfully detected tyrosinase and emitted red fuorescence in melanoma cells and melanocytes. Fluorescence was also observable in zebrafsh and on the skin of guinea pigs when using the RBFP. In mouse and human cells, the RBFP showed good selectivity to tyrosinase. Moreover, in the case of decreased tyrosinase levels or activity caused by siRNAs, kojic acid, or PTU, the probe was sensitive to these changes. Further, the RBFP showed no toxic efects at concentrations of<20?mol/L, both invitro and invivo. Conclusions Our fndings indicate the value and limitations of the RBFP in tyrosinase detection, but suggest the need for further improvement of fuorescent probes in the diagnosis of skin pigmentation disorders.

中南大学湘雅三医院曾庆海、陈静等 | 一种基于试卤灵的酪氨酸酶荧光探针在色素代谢异常性皮肤病模型中的诊断应用评估

本研究论文聚焦小分子荧光探针在色素代谢异常性皮肤病诊断中的价值评估。白癜风、黄褐斑等色素代谢异常性皮肤病的早期诊断十分重要,而酪氨酸酶水平和活性异常可能是早期诊断的可靠指标。一些基于试卤灵的小分子荧光探针(RBFPs)已应用于肿瘤组织酪氨酸酶检测,但尚未运用于色素代谢异常性皮肤病的诊断。本研究使用多种类型的皮肤细胞、斑马鱼、豚鼠和SD鼠作为研究模型,并使用小干扰RNA、曲酸和苯基硫脲(PTU)抑制酪氨酸酶的表达或活性,对一种RBFPs进行综合评估。研究结果发现该RBFPs在细胞和动物模型中均可准确反映酪氨酸酶的表达和活性变化,且具有良好的安全性。因此研究认为RBFPs在色素代谢异常性皮肤病诊断中有一定的价值,但也易受到毛发等因素的干扰,在实际应用中存在局限性,后续研究可针对性改良。

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