A 1,8-naphthalimide-based fluorescent probe for selective and sensitive detection of peroxynitrite and its applications in living cell imaging
A 1,8-naphthalimide-based fluorescent probe for selective and sensitive detection of peroxynitrite and its applications in living cell imaging
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一种基于1,8-萘酰亚胺的荧光探针，用于选择性、灵敏地检测过亚硝酸盐及其在活细胞成像中的应用
DOI：
10.1039/c7ra04317a
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发表时间：
2017-01-01
期刊：
影响因子：
3.9
通讯作者：
Liu, Yangping
中科院分区：
文献类型：
--
作者：
Li, Xiulan;Hou, Jingli;Liu, Yangping
过氧亚硝酸盐（ONOO-）是一种活性氧，可破坏细胞内的多种生物分子，最终导致线粒体功能障碍和细胞死亡。该物种的检测和成像对于了解其在生物过程中的作用具有重要意义。然而，目前的方法大多受到低灵敏度和/或特异性的限制。在本工作中，我们设计并合成了一种新的基于1，8-萘酰亚胺的ONOO-的开启荧光探针NP，其中1，8-萘酰亚胺荧光团直接与甲基（4-羟基苯基）氨基缀合。NP与ONOO-的反应导致甲基（4-羟基苯基）氨基的氧化N-脱芳基化并产生高荧光产物NP-P。在磷酸盐缓冲液中，NP对ONOO-表现出高灵敏度，检测限为11 nM，并且与其他反应物种相比具有高特异性。此外，NP用于特异性检测在LPS和IFN-g刺激后RAW 264.7鼠巨噬细胞中产生的ONOO。我们目前的研究揭示了ONOO-的新探针的设计，由此产生的探针NP具有很大的潜力，使选择性和特异性检测和成像的ONOO（-）在不同的生物和化学系统。
Peroxynitrite (ONOO-) is an extremely reactive oxygen species which can damage a wide array of biomolecules in cells and eventually result in mitochondrial dysfunction and cell death. Detection and imaging of this species is of significant importance to understand its role in biological processes. However, current methods are mostly limited by low sensitivity and/ or specificity. In this work, we designed and synthesized a new 1,8-naphthalimide-based turn-on fluorescent probe NP for ONOO- in which the 1,8-naphthalimide fluorophore was directly conjugated with a methyl(4-hydroxyphenyl) amino group. The reaction of NP with ONOO- led to the oxidative N-dearylation of the methyl(4hydroxyphenyl) amino group and produced a highly fluorescent product NP-P. The probe NP exhibited high sensitivity for ONOO- with a detection limit of 11 nM and high specificity over other reactive species in phosphate buffer. Moreover, NP was used to specifically detect ONOO- generated in RAW264.7 murine macrophages upon stimulation by LPS and IFN-g. Our present study sheds light on the design of new probes for ONOO- and the resulting probe NP has great potential in enabling selective and specific detection and imaging of ONOO (-) in different biological and chemical systems.