Hu Maomao, Wang Boyang, Zhang Hongdan, Wang Han, Li Huixin, Zhang Xinyu, Zhang Jinjin, Lu Qianrun, Fang Guiqian, Wang Juan, Dong Bo
Department of Cardiology, Shandong Provincial Hospital, Shandong University, Jinan, China.
Department of Cardiology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, China.
Front Pharmacol. 2022 Jun 1;13:871059. doi: 10.3389/fphar.2022.871059. eCollection 2022.
Mitochondria are the main sites of reactive sulfur species (RSS) production in living cells. RSS in mitochondria play an important role in physiological and pathological processes of life. In this study, a dual-labeling probe that could simultaneously label the mitochondrial membrane and matrix was designed to quantitatively detect RSS of mitochondria in living cells using nano-level super-resolution imaging. A fluorescent probe CPE was designed and synthesized. The cytotoxicity of CPE was determined and co-localization of CPE with a commercial mitochondrial probe was analyzed in HeLa cells. Then, the uptake patterns of CPE in HeLa cells at different temperatures and endocytosis levels were investigated. The staining characteristics of CPE under different conditions were imaged and quantitated under structured illumination microscopy. A fluorescence probe CPE reacting to RSS was developed, which could simultaneously label the mitochondrial membrane with green fluorescence and the mitochondrial matrix with red fluorescence. CPE was able to demonstrate the mitochondrial morphology and detect the changes of RSS in mitochondria. With the increase of mitochondrial RSS concentration, the light of the red matrix will be quenched. CPE provides a strategy for the design of probes and an attractive tool for accurate examination to changes of mitochondrial morphology and RSS in mitochondria in living cells at the nanoscale.
线粒体是活细胞中活性硫物种（RSS）产生的主要场所。线粒体中的RSS在生命的生理和病理过程中发挥着重要作用。在本研究中，设计了一种能够同时标记线粒体膜和基质的双标记探针，用于利用纳米级超分辨率成像定量检测活细胞中线粒体的RSS。设计并合成了荧光探针CPE。测定了CPE的细胞毒性，并在HeLa细胞中分析了CPE与市售线粒体探针的共定位情况。然后，研究了不同温度和内吞水平下CPE在HeLa细胞中的摄取模式。在结构光照显微镜下对不同条件下CPE的染色特性进行成像和定量分析。开发了一种对RSS有反应的荧光探针CPE，它可以用绿色荧光同时标记线粒体膜，用红色荧光标记线粒体基质。CPE能够显示线粒体形态并检测线粒体中RSS的变化。随着线粒体RSS浓度的增加，红色基质的光将被淬灭。CPE为探针设计提供了一种策略，是在纳米尺度上精确检测活细胞中线粒体形态和线粒体中RSS变化的有吸引力的工具。
