Liu Yanju, Li Manman, Wong Keith Man-Chung, Tong Yan, Yang Huaixia, Kong Jinming
Pharmacy College, Henan University of Chinese Medicine, Zhengzhou 450008, China.
Department of Chemistry, Southern University of Science and Technology, Shenzhen 518055, China.
Int J Anal Chem. 2019 Apr 9;2019:7536431. doi: 10.1155/2019/7536431. eCollection 2019.
In view of the vital role of biothiols in many physiological processes, the development of simple and efficient probe for the detection of biothiols is of great medical significance. In this work, we demonstrate the use of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), which respond rapidly to biothiols especially to glutathione, as a new fluorescent probe for the selective detection and bioimaging of biothiols. This new fluorescent probe can distinguish glutathione from cysteine and homocysteine easily under physiological concentration and detect glutathione quickly within three minutes. This probe exhibits high selectivity to biothiols and the detection limit was determined to be 3.08 × 10 M for glutathione, 8.55 × 10 M for cysteine, and 2.17 × 10 M for homocysteine, respectively. The sensing mechanism was further explored by density functional theory (DFT) and nuclear magnetic resonance (NMR) experiment; results showed that the interaction forces between the probe and biothiols were electrostatic interaction. In addition, the probe has been successfully applied to the detection of biothiols in Eca9706 cells by fluorescence confocal imaging technology.
鉴于生物硫醇在许多生理过程中发挥的重要作用，开发简单高效的生物硫醇检测探针具有重大医学意义。在本研究中，我们展示了使用2,3-二氯-5,6-二氰基-1,4-苯醌（DDQ）作为一种新型荧光探针用于生物硫醇的选择性检测和生物成像，该探针能快速响应生物硫醇，尤其是谷胱甘肽。这种新型荧光探针在生理浓度下能够轻松区分谷胱甘肽与半胱氨酸和同型半胱氨酸，并能在三分钟内快速检测谷胱甘肽。该探针对生物硫醇表现出高选择性，其对谷胱甘肽、半胱氨酸和同型半胱氨酸的检测限分别为3.08×10⁻⁷M、8.55×10⁻⁷M和2.17×10⁻⁷M。通过密度泛函理论（DFT）和核磁共振（NMR）实验进一步探究了传感机制；结果表明探针与生物硫醇之间的相互作用力为静电相互作用。此外，该探针已通过荧光共聚焦成像技术成功应用于Eca9706细胞中生物硫醇的检测。
