A FRET-based ratiometric fluorescence sensor for peroxynitrite detection and its application in living cells, zebrafish, and ultraviolet-induced mouse models imaging

Yu Pan; Xia Gu; Yalu Yuan; Xin Hu; Zepeng Fu; Tianlun Wang; Fengjian Tan; Yongbo Peng; Aijun Chen

doi:10.1016/j.saa.2025.126610

A FRET-based ratiometric fluorescence sensor for peroxynitrite detection and its application in living cells, zebrafish, and ultraviolet-induced mouse models imaging

Spectrochim Acta A Mol Biomol Spectrosc. 2025 Jun 24:343:126610. doi: 10.1016/j.saa.2025.126610. Online ahead of print.

Authors

Yu Pan¹, Xia Gu², Yalu Yuan³, Xin Hu³, Zepeng Fu³, Tianlun Wang³, Fengjian Tan³, Yongbo Peng⁴, Aijun Chen⁵

Affiliations

¹ Department of Dermatology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, PR China.
² Department of Neurology, Weiyuan County People's Hospital, Neijiang, Sichuan 642450, PR China.
³ The Key Laboratory of Biochemistry and Molecular Pharmacology, Chongqing Key Laboratory for Pharmaceutical Metabolism Research, College of Pharmacy, Chongqing Medical University, Chongqing 400016, PR China.
⁴ The Key Laboratory of Biochemistry and Molecular Pharmacology, Chongqing Key Laboratory for Pharmaceutical Metabolism Research, College of Pharmacy, Chongqing Medical University, Chongqing 400016, PR China. Electronic address: pengyongbo2021@cqmu.edu.cn.
⁵ Department of Dermatology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, PR China. Electronic address: chenaijun@hospital.cqmu.edu.cn.

PMID: 40580672
DOI: 10.1016/j.saa.2025.126610

Abstract

Peroxynitrite (ONOO^-) is not only a crucial ROS but also a significant RNS, and its abnormal expression level in vivo is substantially associated with the physiological and pathological outbreak of many diseases. Herein, a ratiometric fluorescence sensor QL-RH was successfully developed using the FRET strategy, in which quinoline and xanthene dye were adopted as the donor and acceptor for the fluorescence resonance energy transfer scaffold, respectively. When QL-RH reacts with ONOO^- by Bayer-Villiger oxidation reaction, and the fluorescence ratiometric response is triggered, the ratio value increases by 50-fold, and it has a good limit of detection (LOD) as low as 28.7 nM, which offered a chemical-specific sensitive molecular tool for the detection of ONOO^-, and the QL-RH of ratiometric characteristics were confirmed by Ultraviolet-Visible and fluorescence spectrophotometer. In addition, QL-RH was also successfully employed for the images of ONOO^- in live cells and zebrafish by LPS stimulation, which increased by ∼9-fold and 8-fold, respectively, and ∼8-fold in mice models irradiated under UV-light, which indicated that QL-RH could become a useful tool for monitoring the process of UV-induced skin aging.

Keywords: FRET strategy; Peroxynitrite anion (ONOO(−)); Ratiometric imaging; UV-induced; Xanthene dye.