Bifunctional Fluorescent Probe for Simultaneous Imaging of H 2 S and Hcy Crosstalk and Therapeutic Evaluation in Acute Kidney Injury

Acute kidney injury (AKI) is a critical clinical condition characterized by the rapid loss of renal function, requiring timely detection and intervention. Hydrogen sulfide (H2S) and homocysteine (Hcy) have been identified as key biomarkers in AKI, yet their dynamic interplay and coordinated regulation remain elusive. Herein, we report a bifunctional fluorescent probe (Probe 1), based on coumarin derivatives, capable of real-time, simultaneous imaging of H2S and Hcy in distinct fluorescence channels with high sensitivity, selectivity, and rapid response. Probe 1 enables the concurrent detection of H2S and Hcy and was successfully applied to visualize both endogenous and exogenous levels in living cells. Notably, this probe achieved the first dual-channel imaging of H2S and Hcy fluctuations in cisplatin-induced AKI models, both in vitro and in vivo. Furthermore, it was employed to evaluate therapeutic responses to AKI-related drug treatments, demonstrating its value for dynamic monitoring of disease progression and intervention efficacy. This study presents the first chemical tool for investigating the synergistic roles of H2S and Hcy in AKI, offering new insights into sulfur-based signaling pathways and providing a promising platform for early diagnosis and therapeutic evaluation in AKI.