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通过比率荧光金碳点监测活细胞中的生物硫醇动力学

Monitoring biothiols dynamics in living cells by ratiometric fluorescent gold carbon dots.

作者信息

Xie Xiaoxue, Peng Zhenqi, Wang Ziqi, Hua Xinyi, Wang Zhifang, Deng Keqin, Yang Xiumei, Huang Haowen, Liu Xuanyong

机构信息

Key Laboratory of Theoretical Organic Chemistry and Function Molecule, Ministry of Education, Hunan Provincial Key Laboratory of Controllable Preparation and Functional Application of Fine Polymers, School of Chemistry and Chemical Engineering, Hunan University of Science and Technology, Xiangtan, 411201, China.

Key Laboratory of Theoretical Organic Chemistry and Function Molecule, Ministry of Education, Hunan Provincial Key Laboratory of Controllable Preparation and Functional Application of Fine Polymers, School of Chemistry and Chemical Engineering, Hunan University of Science and Technology, Xiangtan, 411201, China.

出版信息

Talanta. 2020 Oct 1;218:121214. doi: 10.1016/j.talanta.2020.121214. Epub 2020 May 25.

Abstract

Changes in biothiols, including glutathione (GSH) and cysteine, are closely related to a variety of diseases and cellular functions. Real-time monitoring intracellular GSH and cysteine dynamics in living cells are important for understanding pathophysiological processes. In this study, a stable heterostructure of dual-emission fluorescent gold carbon dots (GCDs) consisting of carbon skeleton and gold nanoclusters was prepared, which emits strong blue fluorescence and weak yellow-orange fluorescence. The blue fluorescence of the as-prepared GCDs has no response to biothiols including GSH and cysteine, but a turn-on yellow-orange fluorescence would be appeared accompanied by the biothiols reacting with the GCDs. Ratiometric fluorescent bioimaging therefore may be established based on the unique GCDs to detect GSH and cysteine levels in living cells. Moreover, real time monitoring GSH and cysteine levels in various cell lines in living cells at different growth stages was realized. The concentration of GSH in cancer cells is higher than that of normal cells, however, the level of cysteine in normal cells is consistently higher than in cancer cells at different stages of cell growth. The fluorescent GCDs probe provides a promising tool for tracking on regulation of GSH and cysteine dynamics by physiological environments due to the capability for real-time quantitation of GSH and cysteine.

摘要

生物硫醇的变化,包括谷胱甘肽(GSH)和半胱氨酸,与多种疾病和细胞功能密切相关。实时监测活细胞内GSH和半胱氨酸的动态变化对于理解病理生理过程至关重要。在本研究中,制备了一种由碳骨架和金纳米团簇组成的具有双发射荧光的稳定异质结构金碳点(GCDs),其发射强蓝色荧光和弱黄橙色荧光。所制备的GCDs的蓝色荧光对包括GSH和半胱氨酸在内的生物硫醇无响应,但当生物硫醇与GCDs反应时会出现开启的黄橙色荧光。因此,可以基于独特的GCDs建立比率荧光生物成像来检测活细胞中GSH和半胱氨酸的水平。此外,还实现了对不同生长阶段活细胞中各种细胞系中GSH和半胱氨酸水平的实时监测。癌细胞中GSH的浓度高于正常细胞,然而,在细胞生长的不同阶段,正常细胞中半胱氨酸的水平始终高于癌细胞。由于能够实时定量GSH和半胱氨酸,荧光GCDs探针为追踪生理环境对GSH和半胱氨酸动态变化的调节提供了一种有前景的工具。

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