The Affiliated Hospital of Southwest Medical University, Luzhou, China.
The Affiliated Hospital of Southwest Medical University, Luzhou, China; Nuclear Medicine and Molecular Imaging Key Laboratory of Sichuan Province, Luzhou, China.
Anal Chim Acta. 2020 Jun 22;1117:18-24. doi: 10.1016/j.aca.2020.03.040. Epub 2020 Mar 24.
This study aimed to develop a novel and practical fluorescent method for GSH detection in complex biological samples. To this end, a series of coumarin-based fluorescent probes was designed and synthesized using various aliphatic halogens as the sensing group. By using a new evaluation method of GSH/Cys/Hcy coexisting conditions, the probe with chloropropionate (CBF3) showed a high selectivity, excellent sensitivity, good stability for GSH detection. The reaction mechanism is proposed as nucleophilic substitution/cyclization and intramolecular charge transfer (ICT), which was confirmed by LC-MS and NMR analysis, as well as density functional theory calculations. In addition, CBF3 was demonstrated to be competent not only for the quantitative detection of GSH in real serum samples, but also for sensing GSH changes in different oxidative stress models in living cells and nematodes. This study showed a practical strategy for constructing GSH-specific fluorescent probes, and provided a sensitive tool for real-time sensing of GSH in real biological samples. The findings would greatly facilitate further investigations on GSH-associated clinical diagnosis and biomedical studies.
本研究旨在开发一种新颖实用的荧光法用于检测复杂生物样品中的 GSH。为此，设计并合成了一系列以香豆素为基础的荧光探针，其中使用了各种脂肪族卤化物作为传感基团。通过使用一种新的 GSH/Cys/Hcy 共存条件的评价方法，具有氯丙酸盐（CBF3）的探针表现出对 GSH 检测的高选择性、优异的灵敏度和良好的稳定性。反应机理被提出为亲核取代/环化和分子内电荷转移（ICT），这通过 LC-MS 和 NMR 分析以及密度泛函理论计算得到了证实。此外，CBF3 不仅能够用于实际血清样品中 GSH 的定量检测，还能够用于在活细胞和线虫中不同氧化应激模型中检测 GSH 的变化。本研究为构建 GSH 特异性荧光探针提供了一种实用策略，并为实时感测实际生物样品中的 GSH 提供了一种灵敏工具。这些发现将极大地促进与 GSH 相关的临床诊断和生物医学研究的进一步发展。
