Li Man, Li Xiang, Chen Linfeng, Li Xiaohong, Liu Chunrong
School of Pharmacy, Hubei University of Science and Technology, Xianning, 437100, PR China; Key Laboratory of Pesticide and Chemical Biology of Ministry of Education, School of Chemistry, Central China Normal University, Wuhan, 430079, PR China; Hubei Engineering Research Center of Traditional Chinese Medicine of South Hubei Province, Xianning, 437100, PR China.
School of Health Science and Engineering, Hubei University, Wuhan 430062, Hubei, PR China.
Talanta. 2025 Mar 1;284:127225. doi: 10.1016/j.talanta.2024.127225. Epub 2024 Nov 15.
Pyruvic acid (PA) is an α-keto acid which exert important biological and pathological functions. The current PA profiling assays are mainly based on the ultraviolet spectroscopy and electrochemical biosensor, requiring killing cells and destroying tissues which limit their application in living cells. Optical imaging provides nondestructive powerful and detective tools to better understand the physiological and pathological role of PA in living systems. However, as far as we know, none of"off - on" PA fluorescent sensor has been developed. Herein, we reported a PA recognition reaction that arylhydroxylamine group could be selectively reduced to acetylamide group by PA. With this recognition reaction, a fluorescence probe (FPA) based on the photoinduced electron transfer (PET) pathway was designed, synthesized and could release strong fluorescence at 447 nm. We proved that FPA could detect PA in aqueous solution, living cells, Caenorhabditis elegans and the roots of Arabidopsis thaliana with good selectivity and sensitivity as low as 0.42 μM. In addition, we successfully using probe FPA to study the intracellular PA production pathway in cells and evaluated its physiological level in Arabidopsis roots at different growth stages. The results show that the physiological level of PA in Arabidopsis thaliana roots is closely associated with their growth stages, which indicated that PA might act as a carbon source and related growth signaling molecule to promote plant growth and root elongation. Therefore, we expect probe FPA to be a powerful tool to better understand the physiological and pathological role of PA.
丙酮酸（PA）是一种α-酮酸，具有重要的生物学和病理学功能。目前的PA分析方法主要基于紫外光谱和电化学生物传感器，需要杀死细胞和破坏组织，这限制了它们在活细胞中的应用。光学成像提供了无损的强大检测工具，以便更好地理解PA在生命系统中的生理和病理作用。然而，据我们所知，尚未开发出“开-关”型PA荧光传感器。在此，我们报道了一种PA识别反应，即芳基羟胺基团可被PA选择性还原为乙酰酰胺基团。基于此识别反应，设计、合成了一种基于光诱导电子转移（PET）途径的荧光探针（FPA），其在447nm处可发出强荧光。我们证明FPA能够在水溶液、活细胞、秀丽隐杆线虫和拟南芥根中检测PA，具有良好的选择性，灵敏度低至0.42μM。此外，我们成功地使用探针FPA研究了细胞内PA的产生途径，并评估了其在拟南芥根不同生长阶段的生理水平。结果表明，拟南芥根中PA的生理水平与其生长阶段密切相关，这表明PA可能作为碳源和相关生长信号分子促进植物生长和根伸长。因此，我们期望探针FPA成为更好地理解PA生理和病理作用的有力工具。
