Development of a Golgi-targeted superoxide anion fluorescent probe for elucidating protein GOLPH3 function in myocardial ischemia-reperfusion injury.

Superoxide anion (O) is an important reactive oxygen species (ROS) and participates in various physiological and pathological processes in the organism. The O burst induced by ischemia-reperfusion (I/R) is associated with cardiovascular disease and promotes the cell apoptosis. In this work, a turn-on type Golgi-targeting fluorescent probe Gol-Cou-O was rationally designed for sensitive and selective detection of O. The minimum detection limit concentration for O was about 3.9 × 10 M in aqueous solution. Gol-Cou-O showed excellent capacity of detecting exogenous and endogenous O in living cells and zebrafish, and was also used to capture the up-regulated O level during the duration of I/R process in cardiomyocytes. Golgi Phosphoprotein 3 (GOLPH3) is a potential Golgi stress marker protein and plays a key role in cells apoptosis during I/R. The fluorescence imaging and flow cytometry assay results indicated that silencing GOLPH3 through siRNA could give rise to the down-regulated O level and alleviation of apoptosis in I/R myocardial cells. Thus, development of Gol-Cou-O provides a diagnostic tool for myocardial oxidative stress injury and distinct insights on roles of GOLPH3 in myocardial I/R injury.