Analytica Chimica Acta ( IF 6.2 ) Pub Date : 2023-03-16 , DOI: 10.1016/j.aca.2023.341100 Ao Li 1 , Yalan Liu 1 , Labapuchi 1 , Zhe Chen 1 , Songjiao Li 1 , Rongbin Zhong 2 , Dan Cheng 2 , Linxi Chen 1 , Longwei He 1
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.
中文翻译:
开发一种高尔基体靶向超氧阴离子荧光探针,用于阐明蛋白质 GOLPH3 在心肌缺血再灌注损伤中的功能
超氧阴离子(O 2 •- )是一种重要的活性氧(ROS),参与机体的多种生理和病理过程。缺血再灌注(I/R) 诱导的O 2 •-爆发与心血管疾病有关并促进细胞凋亡。本工作合理设计了一种开启型高尔基体靶向荧光探针Gol-Cou-O 2 •- ,用于O 2 •-的灵敏和选择性检测。O 2 •-的最低检测限浓度在水溶液中约为3.9 × 10 -7 M。Gol-Cou-O 2 •-在活细胞和斑马鱼中显示出检测外源性和内源性 O 2 •-的出色能力,并且还用于在心肌细胞 I/R 过程期间捕获上调的 O 2 •-水平。高尔基体磷蛋白 3 (GOLPH3) 是一种潜在的高尔基体应激标记蛋白,在 I/R 期间的细胞凋亡中起着关键作用。荧光成像和流式细胞术检测结果表明,通过siRNA 沉默GOLPH3 可导致I/R 心肌细胞O 2 •-水平下调和细胞凋亡减轻。因此,开发Gol-Cou-O 2 •-提供了心肌氧化应激损伤的诊断工具和对 GOLPH3 在心肌 I/R 损伤中作用的独特见解。