Sadek Maiada M, Barzegar Amiri Olia Mina, Nowell Cameron J, Barlow Νicholas, Schiesser Carl H, Nicholson Sandra E, Norton Raymond S
Medicinal Chemistry, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria 3052, Australia.
School of Chemistry and Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Victoria 3010, Australia.
Bioorg Med Chem. 2017 Oct 15;25(20):5743-5748. doi: 10.1016/j.bmc.2017.08.054. Epub 2017 Sep 5.
Nitric oxide (NO) is an important effector molecule in host defence against bacterial pathogens. The development of fluorescence imaging to monitor NO production in vitro and in vivo will increase our understanding of its biological role. Recently, a novel 'trappable' fluorescent blue 'turn-on' Cu(II)-complexed coumarin-based probe (CB) has been developed to detect NO. In this study, CB was investigated to evaluate its ability to detect NO in macrophages. Using confocal microscopy, NO was successfully detected in macrophages in the presence of stimuli that induce nitric oxide synthase (iNOS), the enzyme responsible for production of NO. The time dependence and subcellular compartmentalisation of CB in macrophages were evaluated. The probe can be trapped within cells and reacts directly and specifically with NO, rendering it a promising tool for imaging NO in response to pharmacological agents that modulate its level, for example during bacterial infections.
一氧化氮（NO）是宿主抵御细菌病原体的重要效应分子。用于监测体外和体内NO生成的荧光成像技术的发展将增进我们对其生物学作用的理解。最近，一种新型的“可捕获”荧光蓝色“开启”型铜（II）络合香豆素基探针（CB）已被开发用于检测NO。在本研究中，对CB检测巨噬细胞中NO的能力进行了研究。使用共聚焦显微镜，在存在诱导一氧化氮合酶（iNOS，负责产生NO的酶）的刺激物的情况下，成功在巨噬细胞中检测到了NO。评估了CB在巨噬细胞中的时间依赖性和亚细胞定位。该探针可被困在细胞内，并直接与NO特异性反应，使其成为一种有前景的工具，用于成像响应调节其水平的药物（例如在细菌感染期间）而产生的NO。
