Chai Xianzhi, Ma Xiuhua, Sun Lu-Lu, Hu Yuqing, Zhang Weijian, Zhang Shiyao, Zhou Jiaqi, Zhu Liangliang, Han Hai-Hao, He Xiao-Peng
School of Environmental and Chemical Engineering, Jiangsu University of Science and Technology, Zhenjiang, Jiangsu 212100, China.
State Key Laboratory of Molecular Engineering of Polymers, Department of Macromolecular Science, Fudan University, Shanghai 200438, China.
Anal Chem. 2024 Dec 24;96(51):20161-20168. doi: 10.1021/acs.analchem.4c03759. Epub 2024 Dec 9.
Mitochondria are the energy factory of cells and can be easily damaged by reactive oxygen species (ROS) because of the frequent occurrence of oxidative stress. Abnormality in mitophagy is often associated with many diseases including inflammation, cancer, and aging. While previously developed fluorescent probes mainly focus on detecting just ROS or mitophagy, quite rare studies have endeavored to comprehensively capture the entire mitophagic process, encompassing both the production of ROS and the induction of mitophagy. Herein, we report a new ratiometric fluorescent probe for tracking peroxynitrite (ONOO) as well as the subsequent oxidative stress-induced mitophagy. To a naphthalimide-based dye, an ONOO-responsive diphenyl phosphinate moiety and the mitochondria-targeting triphenylphosphonium group were attached. The probe showed a highly selective response to ONOO through an addition-elimination reaction with diphenyl phosphinate. Owing to its outstanding pH stability and organelle-targeting ability, was successfully used to detect mitophagy induced by oxidative stress after the generation of ONOO. In the meantime, the probe was also used to track starvation-induced mitophagy and indicate that starvation-induced mitophagy is independent of ONOO. Therefore, has the ability to precisely track oxidative stress-induced mitophagy by distinguishing it from starvation-induced mitophagy. This study offers a new chemical tool to study the relationship between ROS generation and mitophagy.
线粒体是细胞的能量工厂，由于氧化应激频繁发生，极易受到活性氧（ROS）的损伤。线粒体自噬异常常与包括炎症、癌症和衰老在内的许多疾病相关。虽然先前开发的荧光探针主要专注于仅检测ROS或线粒体自噬，但很少有研究致力于全面捕捉整个线粒体自噬过程，包括ROS的产生和线粒体自噬的诱导。在此，我们报告了一种新型比率荧光探针，用于追踪过氧亚硝酸盐（ONOO）以及随后氧化应激诱导的线粒体自噬。在基于萘二甲酰亚胺的染料上连接了一个对ONOO有响应的二苯基次膦酸酯部分和线粒体靶向三苯基鏻基团。该探针通过与二苯基次膦酸酯的加成-消除反应对ONOO表现出高度选择性响应。由于其出色的pH稳定性和细胞器靶向能力，该探针成功用于检测ONOO产生后氧化应激诱导的线粒体自噬。同时，该探针还用于追踪饥饿诱导的线粒体自噬，并表明饥饿诱导的线粒体自噬与ONOO无关。因此，该探针能够通过将氧化应激诱导的线粒体自噬与饥饿诱导的线粒体自噬区分开来，精确追踪氧化应激诱导的线粒体自噬。本研究提供了一种新的化学工具来研究ROS产生与线粒体自噬之间的关系。
