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

• Published in: Analytica chimica acta 2023-05, Vol.1255, p.341100-341100, Article 341100
• Main Authors: Li, Ao, Liu, Yalan, Labapuchi, Chen, Zhe, Li, Songjiao, Zhong, Rongbin, Cheng, Dan, Chen, Linxi, He, Longwei
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis; Fluorescent Dyes - metabolism; Fluorescent Dyes - toxicity; Fluorescent probe; Golgi apparatus; Golgi Apparatus - metabolism; GOLPH3; Myocardial ischemia-reperfusion injury; Myocardial Reperfusion Injury - diagnostic imaging; Myocardial Reperfusion Injury - pathology; Oxygen - metabolism; Reactive Oxygen Species - metabolism; Superoxide anion; Superoxides; Zebrafish
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2023.341100

Superoxide anion (O2•-) is an important reactive oxygen species (ROS) and participates in various physiological and pathological processes in the organism. The O2•- 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-O2•- was rationally designed for sensitive and selective detection of O2•-. The minimum detection limit concentration for O2•- was about 3.9 × 10−7 M in aqueous solution. Gol-Cou-O2•- showed excellent capacity of detecting exogenous and endogenous O2•- in living cells and zebrafish, and was also used to capture the up-regulated O2•- 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 O2•- level and alleviation of apoptosis in I/R myocardial cells. Thus, development of Gol-Cou-O2•- provides a diagnostic tool for myocardial oxidative stress injury and distinct insights on roles of GOLPH3 in myocardial I/R injury. Engineering a Golgi-targeted superoxide anion fluorescent probe Gol-Cou-O2•- for investigating the function of protein GOLPH3 in myocardial ischemia-reperfusion injury. [Display omitted] •A Golgi-targeting O2.•- fluorescent probe Gol-Cou-O2•- was rationally developed.•Probe was used to monitor O2.•- burst during I/R process in cardiomyocytes.•Probe could detect the O2.•- level changes and apoptosis alleviation by silencing GOLPH3.•The function of GOLPH3 during myocardial I/R injury was elucidated for the first time.