Rational Design of a Novel Dual-Functional Fluorescent Probe for Simultaneous Monitoring and Imaging of Mitochondrial CO and ATP Fluctuations during Drug-Induced Liver Injury without Spectral Crosstalk

Drug-induced liver injury (DILI) poses a terrible threat to human health. Carbon monoxide (CO) is documented to have hepatoprotective effects by relieving hepatic oxidative stress caused by DILI, and adenosine-5'-triphosphate (ATP) has been considered a vital biomarker for reflecting mitochondrial energy metabolism in DILI. However, the dynamic fluctuations and cooperative actions of CO and ATP in DILI remain unclear. Herein, a new fluorescent probe CCRD for the simultaneous detection of CO and ATP fluctuations in two well-separated emission channels without signal crosstalk was constructed. The nitro group on the coumarin unit of CCRD after reacting with CO, was selectively reduced to its amino form, affording a distinct blue emission at 448 nm. By comparison, the coordination of ATP with CCRD caused the ring opening of the rhodamine moiety, generating an intense red emission at 584 nm. The probe CCRD enabled dual-channel imaging of endogenous and exogenous CO and ATP within HepG2 cells. Furthermore, the synergistic dynamic changes of CO bursts and ATP consumption in live cells and mouse livers triggered by DILI were monitored for the first time by using CCRD. This work not only realizes the bidirectional channel imaging of CO and ATP in vivo and in vitro but also provides a powerful tool for the early diagnosis and therapy of DILI.