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

doi:10.19894/j.issn.1000-0518.220308

Chinese Journal of Applied Chemistry ›› 2023, Vol. 40 ›› Issue (4): 500-508.DOI: 10.19894/j.issn.1000-0518.220308

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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.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)

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Abstract

Abstract:

An “off-on” fluorescence probe （4a） based on N，N-（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

CLC Number:

O657.3

Hong-Li YU, Si-Yi ZHOU, Chen HONG, Wen LUO. A Flavone-based “Off-On” Fluorescence Probe for Detecting Butyrylcholinesterase in Living Cell[J]. Chinese Journal of Applied Chemistry, 2023, 40(4): 500-508.

Figures/Tables 10

Fig.1 Chemical structures of some reported BChE probes （A-E） and flavone probes （F-J）

Fig.2 Synthetic route of compounds 4a and 4b

Fig.3 Absorption （A） and fluorescent spectra （B） of probe 4a （10 μmol/L） with or without BchE （43.6 U/mL） in PBS

Fig.4 A plot of fluorescence intensity at 574 nm vs the reaction time for the probe 4a （10 μmol/L） in PBS in the presence of different amounts of BChE （λex= 430 nm， slit： 2.0/0.5 nm）

Fig.5 The fluorescence change multiple of probe 4a （10 μmol/L） toward various analytes in PBS

Fig.6 Fluorescence intensity （574 nm） of probe 4a （10 μmol/L） with BChE （43.6 U/mL） and the interference of coexisting various small molecules （50 μmol/L） in PBS

Fig. 7 Effects of pH value on the fluorescence intensity （574 nm） of probe 4a （10 μmol/L） with or without BChE （43.6 U/mL）

Fig.8 The recognition mechanism of probe 4a for BChE

Fig.9 MTT assay of probe 4a and HepG2 cells after 24 h incubation

Fig.10 Response of BChE by probe 4a （10 μmol/L） in living HepG2 cells pretreated without （A-C） or with tacrine （D-F）.（A）.（D） dark field，（B）.（E） bright field，（C）.（F） merged

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

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