A “Cocktail” Fluorescent Probe for Multi‐ROS Imaging Unveils Ferroptosis‐Driven Liver Fibrosis Development

Abstract Liver fibrosis is a pathological repair response to chronic liver injury and may progress to cirrhosis, liver failure, or hepatocellular carcinoma if untreated. Currently, no approved therapies specifically target advanced liver fibrosis, thus exploring the molecular mechanisms underlying liver fibrosis has become crucial. Previous studies have highlighted significant controversy regarding the role of ferroptosis in liver fibrosis. Given that reactive oxygen species (ROS) serve as key mediators to both processes, ROS may serve as a molecular nexus connecting ferroptosis and liver fibrosis. To comprehensively elucidate the molecular network involving ROS and ferroptosis in liver fibrosis, we designed and synthesized the first multi‐functional “cocktail” fluorescence probe, FP‐ROS, enabling highly sensitive and selective simultaneous imaging of O 2 •− , H 2 O 2 , and ONOO − . FP‐ROS was successfully employed to assess ferroptosis levels in the livers of fibrosis mice following drug intervention. Combining transcriptomic and proteomic analyses, we elucidated the signaling pathway NOX→ONOO − →GCLM(C46)→GSH→ferroptosis→hepatic stellate cells (HSCs) activation. This study demonstrates that ferroptosis plays a critical role in HSCs activation and further elucidates the molecular interplay between ROS and ferroptosis in fibrosis progression. These findings provide novel insights into the diagnosis and therapeutic strategies for liver fibrosis.