School of Pharmacy, Fudan University, Shanghai, 201203, PR China.
School of Pharmacy, Fudan University, Shanghai, 201203, PR China.
Anal Chim Acta. 2019 Aug 27;1067:88-97. doi: 10.1016/j.aca.2019.03.048. Epub 2019 Mar 23.
Nitric oxide (NO), a ubiquitous gasotransmitter which plays critical roles in cardiovascular, nervous, and immune systems related diseases, is closely related in the physiological and pathological processes of mitochondria and lysosomes. Thus, monitoring NO in mitochondria or lysosomes is very meaningful for NO related chemical biology. Herein, we rationally designed four NO probes, BDP-NO, Mito-NO-T, Mito-NO and Lyso-NO, based on BODIPY dye substituted at meso position with 5-amino-2-methoxy-phenyl scaffold. These four probes all showed fast fluorescence off-on response toward NO with excellent selectivity and high sensitivity with the detection limit of BDP-NO to reach 5.7 nM. We introduced triphenylphosphonium and morpholine moieties onto BODIPY scaffold respectively to enable organelle-targetability. MTT and flow cytometry assay demonstrated that the probes exhibited low cytotoxicity, which was beneficial to the biological application in living cells. Confocal fluorescence microscopy experiments confirmed excellent mitochondria targeting for Mito-NO and lysosome-targeting with Lyso-NO for the detection of NO in living cells.
一氧化氮（NO）是一种普遍存在的气体递质，在心血管、神经和免疫系统相关疾病中发挥着关键作用，它与线粒体和溶酶体的生理和病理过程密切相关。因此，监测线粒体或溶酶体中的 NO 对于与 NO 相关的化学生物学非常有意义。在这里，我们基于 5-氨基-2-甲氧基-苯基取代的 BODIPY 染料，合理设计了四个 NO 探针，BDP-NO、Mito-NO-T、Mito-NO 和 Lyso-NO。这四个探针都对 NO 表现出快速的荧光开-关响应，具有优异的选择性和高灵敏度，BDP-NO 的检测限达到 5.7 nM。我们分别在 BODIPY 支架上引入三苯基膦和吗啉基团，以实现细胞器靶向性。MTT 和流式细胞术测定表明，这些探针具有低细胞毒性，有利于在活细胞中的生物学应用。共聚焦荧光显微镜实验证实了 Mito-NO 对线粒体的优异靶向性和 Lyso-NO 对溶酶体的靶向性，用于检测活细胞中的 NO。
