Ouyang Jie, Hong Hao, Shen Chao, Zhao Yong, Ouyang Chenguang, Dong Lei, Zhu Jianhui, Guo Zijian, Zeng Ke, Chen Jiangning, Zhang Chenyu, Zhang Junfeng
The School of Chemistry and Chemical Engineering, Tianjin University of Technology, Tianjin 300191, China.
Free Radic Biol Med. 2008 Nov 15;45(10):1426-36. doi: 10.1016/j.freeradbiomed.2008.08.016. Epub 2008 Aug 28.
Fluorescence imaging of nitric oxide (NO) in vitro and in vivo is essential to developing our understanding of the role of nitric oxide in biology and medicine. Current probes such as diaminofluorescein depend on reactions with oxidized NO products, but not with nitric oxide directly, and this limits their applicability. Here we report the formation of an imaging probe for nitric oxide by coordinating the highly fluorescent chemical 4-methoxy-2-(1H-naphtho[2,3-d]imidazol-2-yl)phenol (MNIP) with Cu(II). The coordination compound MNIP-Cu reacts rapidly and specifically with nitric oxide to generate a product with blue fluorescence that can be used in vitro and in vivo. In the present study MNIP-Cu was used to reveal nitric oxide produced by inducible nitric oxide synthase in lipopolysaccharide (LPS)-activated macrophages (Raw 264.7 cells) and by endothelial nitric oxide synthase in endothelial cells (HUVEC). MNIP-Cu was also used to evaluate the distribution of nitric oxide synthesis in a model of acute liver injury induced by LPS and d-galactosamine in mice. The results demonstrate that MNIP-Cu can act as a novel fluorescent probe for nitric oxide and has many potential applications in biomedical research.
体外和体内一氧化氮（NO）的荧光成像对于深入了解一氧化氮在生物学和医学中的作用至关重要。目前的探针，如二氨基荧光素，依赖于与一氧化氮氧化产物的反应，而不是直接与一氧化氮反应，这限制了它们的适用性。在此，我们报告了通过将高荧光化学物质4-甲氧基-2-(1H-萘并[2,3-d]咪唑-2-基)苯酚（MNIP）与Cu(II)配位形成一种用于一氧化氮的成像探针。配位化合物MNIP-Cu与一氧化氮迅速且特异性地反应，生成具有蓝色荧光的产物，可用于体外和体内。在本研究中，MNIP-Cu用于揭示脂多糖（LPS）激活的巨噬细胞（Raw 264.7细胞）中诱导型一氧化氮合酶产生的一氧化氮以及内皮细胞（HUVEC）中内皮型一氧化氮合酶产生的一氧化氮。MNIP-Cu还用于评估LPS和d-半乳糖胺诱导的小鼠急性肝损伤模型中一氧化氮合成的分布。结果表明，MNIP-Cu可作为一种新型的一氧化氮荧光探针，在生物医学研究中有许多潜在应用。
