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

Anal Chim Acta. 2023 May 15:1255:341100. doi: 10.1016/j.aca.2023.341100. Epub 2023 Mar 16.

Abstract

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.

Keywords: Apoptosis; Fluorescent probe; GOLPH3; Golgi apparatus; Myocardial ischemia-reperfusion injury; Superoxide anion.

MeSH terms

  • Animals
  • Fluorescent Dyes / metabolism
  • Fluorescent Dyes / toxicity
  • Golgi Apparatus / metabolism
  • Myocardial Reperfusion Injury* / diagnostic imaging
  • Myocardial Reperfusion Injury* / pathology
  • Oxygen / metabolism
  • Reactive Oxygen Species / metabolism
  • Superoxides*
  • Zebrafish

Substances

  • Superoxides
  • Fluorescent Dyes
  • Reactive Oxygen Species
  • Oxygen