Visualizing Peroxynitrite Fluxes in Endothelial Cells Reveals the Dynamic Progression of Brain Vascular Injury
Visualizing Peroxynitrite Fluxes in Endothelial Cells Reveals the Dynamic Progression of Brain Vascular Injury
复制标题
内皮细胞中过氧亚硝酸盐通量的可视化揭示了脑血管损伤的动态进展
DOI：
10.1021/jacs.5b06865
复制
发表时间：
2015-09-30
影响因子：
15
通讯作者：
Hu, You-Hong
中科院分区：
文献类型：
--
作者：
Li, Xin;Tao, Rong-Rong;Hu, You-Hong
越来越多的证据表明，过氧亚硝基阴离子（ONOO-）在脑血管系统中的形成有助于缺血性损伤的进展，而潜在的分子机制仍然是难以捉摸的。为了全面了解ONOO-生物学，实现内源ONOO-通量实时追踪的有效工具是必不可少的。虽然已经报道了一些ONOO-荧光探针，但直接观察活小鼠脑血管中的ONOO-通量仍然是一个挑战。在此，我们提出了一种用于ONOO成像的荧光开关探针（NP 3）。NP 3在体外和体内均表现出对ONOO的良好特异性、快速响应和高灵敏度。此外，NP 3是双光子可激发的，并且容易穿透血脑屏障。这些所需的生物物理和药代动力学特性赋予NP 3以优异的时间和空间分辨率监测损伤后脑血管ONOO生成的能力。作为概念的证明，NP 3使得能够通过使用双光子激光扫描显微镜直接可视化活小鼠脑中缺血进展中的神经血管ONOO形成。由于这些有利的特性，NP 3在体外和体内的各种病理生理学进展中可视化内源性过氧亚硝酸盐通量方面具有很大的前景。
Accumulating evidence suggests that formation of peroxynitrite (ONOO-) in the cerebral vasculature contributes to the progression of ischemic damage, while the underlying molecular mechanisms remain elusive. To fully understand ONOO- biology, efficient tools that can realize the real-time tracing of endogenous ONOO- fluxes are indispensable. While a few ONOO- fluorescent probes have been reported, direct visualization of ONOO- fluxes in the cerebral vasculature of live mice remains a challenge. Herein, we present a fluorescent switch-on probe (NP3) for ONOO- imaging. NP3 exhibits good specificity, fast response, and high sensitivity toward ONOO- both in vitro and in vivo. Moreover, NP3 is two-photon excitable and readily blood-brain barrier penetrable. These desired photophysical and pharmacokinetic properties endow NP3 with the capability to monitor brain vascular ONOO- generation after injury with excellent temporal and spatial resolution. As a proof of concept, NP3 has enabled the direct visualization of neurovascular ONOO- formation in ischemia progression in live mouse brain by use of two-photon laser scanning microscopy. Due to these favorable properties, NP3 holds great promise for visualizing endogenous peroxynitrite fluxes in a variety of pathophysiological progressions in vitro and in vivo.