应用化学 ›› 2023, Vol. 40 ›› Issue (4): 500-508.DOI: 10.19894/j.issn.1000-0518.220308

• 研究论文 • 上一篇    下一篇

基于黄酮骨架的“关-开”型荧光探针用于检测活细胞内丁酰胆碱酯酶

于红丽1,2, 周思仪2, 洪琛1(), 罗稳2()   

  1. 1.河南大学淮河医院,开封 475000
    2.河南大学天然药物与免疫工程重点实验室,开封 475004
  • 收稿日期:2022-09-20 接受日期:2023-02-14 出版日期:2023-04-01 发布日期:2023-04-17
  • 通讯作者: 洪琛,罗稳
  • 基金资助:
    河南省科技攻关计划项目(212102311030)

A Flavone-based “Off-On” Fluorescence Probe for Detecting Butyrylcholinesterase in Living Cell

Hong-Li YU1,2, Si-Yi ZHOU2, Chen HONG1(), Wen LUO2()   

  1. 1.Huaihe Hosipital,Henan University,Kaifeng 475000,China
    2.Key Laboratory of Natural Medicine and Immuno-Engineering,Henan University,Kaifeng 475004,China
  • Received:2022-09-20 Accepted:2023-02-14 Published:2023-04-01 Online:2023-04-17
  • Contact: Chen HONG,Wen LUO
  • About author:luowen83@163.com
    hongchenbest@163.com
  • Supported by:
    the Science and Technology Planning Project of Henan Province(212102311030)

摘要:

以间苯二酚为起始原料,设计合成了一种基于NN-二甲胺基萘取代的黄酮母核(3)为荧光报告基团的“关-开”型丁酰胆碱酯酶(BChE)荧光探针(4a),并通过核磁共振波谱(NMR)和高分辨质谱(HRMS)进行了结构表征。结果表明,探针4a能够快速地对BChE产生特异性响应,在pH=8.0的磷酸盐缓冲液(PBS)中,574 nm处荧光强度明显增强,同时肉眼可见溶液颜色的改变。通过高效液相色谱(HPLC)和HRMS对响应机理进行了探讨,发现BChE可使探针的酯键发生断裂,生成具有强荧光的黄酮母核(3)。此外,探针4a可用于检测人肝癌细胞(HepG2)中的BChE并进行荧光成像。

关键词: 黄酮, 丁酰胆碱酯酶, 荧光探针

Abstract:

An “off-on” fluorescence probe (4a) based on NN-(dimethylamino)naphthalenyl flavone (3) is designed and synthesized for detection of butyrylcholinesterase (BChE) by using resorcinol as starting materials. The structure of the probe is identified by nuclear magnetic resonance spectroscopy (NMR) and high resolution mass spectrometry (HRMS). The results show that probe 4a could react rapidly and specifically with BChE, and its fluorescence intensity increases obviously at 574 nm in PBS of pH 8.0. Meanwhile, the color change of the solution can be seen by naked eyes. The response mechanism is explored through high performance liquid chromatography (HPLC) and HRMS. The results reveal that BChE catalyzes the cleavage of the ester bond, and strongly fluorescent flavone 3 is released. In addition, probe 4a could be used for fluorescence imaging of BChE in living human hepatocellular carcinoma (HepG2) cells.

Key words: Flavone, Butyrylcholinesterase, Fluorescence probe

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