一种溶酶体靶向荧光探针,用于药物分析和多种生物系统中的硫醇检测。
A lysosome-targeted fluorescent probe for thiol detection in drug analysis and multiple biological systems.
发表日期:2024 Aug 28
作者:
Yitong Liu, Juan Song, Yan Li, Peng Hou, Haijun Wang, Jiaming Wang, Chuan He, Song Chen
来源:
ANTIOXIDANTS & REDOX SIGNALING
摘要:
生物硫醇以其独特的巯基(-SH)基团为特征,具有优异的抗氧化特性,可有效中和生物体中活性氧(ROS)对细胞结构的损害。此外,溶酶体通过其内部酶的作用,在分解受损生物分子、调节细胞氧化还原状态和减轻氧化应激方面发挥着至关重要的作用。为了促进细胞内生物硫醇的快速监测,特别是在溶酶体内,我们在本研究中构建了一种靶向溶酶体的生物硫醇荧光探针 PHL-DNP。 PHL-DNP 在水性测试系统中表现出优异的光物理性质,包括强荧光增强响应、优异的选择性和低检测限(Cys 16.5 nM、Hcy 16.8 nM、GSH 21.3 nM、Cap 26.6 nM)。这些特性使得能够轻松高效地对试纸条上的半胱氨酸进行鉴定,并准确测定卡托普利片剂的有效含量。值得注意的是,PHL-DNP 表现出低细胞毒性和精确的溶酶体靶向性。通过生物成像,PHL-DNP不仅监测氧化应激下生物硫醇水平的变化,还评估了复杂生物系统(如活HeLa细胞、斑马鱼、肿瘤组织切片和萝卜根)中的生物硫醇。这为生物硫醇的定量分析、疾病标志物检测和药物测试提供了一种有前景的工具。© 2024。作者获得 Springer-Verlag GmbH(Springer Nature 旗下德国公司)的独家许可。
Biothiols, characterized by their unique sulfhydryl (-SH) groups, possess excellent antioxidant properties, effectively neutralizing the damage to cellular structures caused by reactive oxygen species (ROS) in living organisms. Additionally, lysosomes play a crucial role in decomposing damaged biomolecules through the action of their internal enzymes, regulating the cellular redox state, and mitigating oxidative stress. To facilitate rapid monitoring of intracellular biothiols, particularly within lysosomes, we constructed a lysosome-targeted biothiol fluorescent probe, PHL-DNP, in this study. PHL-DNP exhibited excellent photophysical properties in an aqueous test system, including strong fluorescence enhancement response, excellent selectivity, and low detection limits (Cys 16.5 nM, Hcy 16.8 nM, GSH 21.3 nM, Cap 26.6 nM). These attributes enabled easy and efficient qualification of Cys on test strips and accurate determination of the effective content of captopril tablets. Notably, PHL-DNP demonstrated low cytotoxicity and precise lysosomal targeting. Through bioimaging, PHL-DNP not only monitored changes in biothiol levels under oxidative stress but also assessed biothiols in complex biological systems such as live HeLa cells, zebrafish, tumor tissue sections, and radish roots. This provides a promising tool for quantitative analysis of biothiols, disease marker detection, and drug testing.© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.