Fluorescent probe for the imaging of superoxide and peroxynitrite during drug-induced liver injury

Luling Wu; Jihong Liu; Xue Tian; Robin R Groleau; Steven D Bull; Ping Li; Bo Tang; Tony D James

doi:10.1039/d0sc05937d

Fluorescent probe for the imaging of superoxide and peroxynitrite during drug-induced liver injury

Chem Sci. 2021 Jan 4;12(11):3921-3928. doi: 10.1039/d0sc05937d.

Authors

Luling Wu^{1

2}, Jihong Liu^{1

2}, Xue Tian², Robin R Groleau², Steven D Bull², Ping Li¹, Bo Tang¹, Tony D James^{1

2

3}

Affiliations

¹ College of Chemistry, Chemical Engineering and Materials Science, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Institutes of Biomedical Sciences, Shandong Normal University Jinan 250014 People's Republic of China lip@sdnu.edu.cn tangb@sdnu.edu.cn.
² Department of Chemistry, University of Bath Bath BA2 7AY UK t.d.james@bath.ac.uk.
³ School of Chemistry and Chemical Engineering, Henan Normal University Xinxiang 453007 P. R. China.

Abstract

Drug-induced liver injury (DILI) is an important cause of potentially fatal liver disease. Herein, we report the development of a molecular probe (LW-OTf) for the detection and imaging of two biomarkers involved in DILI. Initially, primary reactive oxygen species (ROS) superoxide (O₂˙^-) selectively activates a near-infrared fluorescence (NIRF) output by generating fluorophore LW-OH. The C[double bond, length as m-dash]C linker of this hemicyanine fluorophore is subsequently oxidized by reactive nitrogen species (RNS) peroxynitrite (ONOO^-), resulting in cleavage to release xanthene derivative LW-XTD, detected using two-photon excitation fluorescence (TPEF). An alternative fluorescence pathway can occur through cleavage of LW-OTf by ONOO^- to non-fluorescent LW-XTD-OTf, which can react further with the second analyte O₂˙^- to produce the same LW-XTD fluorescent species. By combining NIRF and TPEF, LW-OTf is capable of differential and simultaneous detection of ROS and RNS in DILI using two optically orthogonal channels. Probe LW-OTf could be used to detect O₂˙^- or O₂˙^- and ONOO^- in lysosomes stimulated by 2-methoxyestradiol (2-ME) or 2-ME and SIN-1 respectively. In addition, we were able to monitor the chemoprotective effects of tert-butylhydroxyanisole (BHA) against acetaminophen (APAP) toxicity in living HL-7702 cells. More importantly, TPEF and NIRF imaging confirmed an increase in levels of both O₂˙^- and ONOO^- in mouse livers during APAP-induced DILI (confirmed by hematoxylin and eosin (H&E) staining).