Simultaneous <i>In Situ</i> Monitoring and Imaging of Superoxide Anion, Hydrogen Peroxide, and Protein Phosphorylation for Early Monitoring of Atherosclerosis

Investigating the correlation between changes in multiple active molecule levels and the progression of atherosclerosis holds significant practical value for achieving early disease monitoring. To this end, this study constructed a fluorescence nanoprobe for monitoring superoxide anion (O 2 •– ), hydrogen peroxide (H 2 O 2 ), and protein phosphorylation, which enabled the synchronous detection and imaging of the three molecules at both the cellular and in vivo levels, thereby systematically exploring the correlation between their level changes and early disease progression. Two-photon fluorescence imaging was used to monitor changes in the levels of the three molecules in the aorta of model, which showed that the levels of protein phosphorylation, O 2 •–, and H 2 O 2 in vascular tissues of the disease group were significantly elevated. More importantly, deep tissue imaging (approximately 140 μm) was achieved utilizing the two-photon imaging. Meanwhile, changes in active molecules in the blood during different disease stages were observed, and as the disease progressed, all three indicators showed a marked increase; among them, hydrogen peroxide exhibits the most pronounced fluorescence response. Serum testing combined with two-photon imaging provides support for early monitoring of atherosclerosis.