A Ratiometric Fluorescent Probe for Biological Signaling Molecule H2S: Fast Response and High Selectivity
A Ratiometric Fluorescent Probe for Biological Signaling Molecule H2S: Fast Response and High Selectivity
复制标题
用于生物信号分子 H2S 的比率荧光探针：快速响应和高选择性
DOI：
10.1002/chem.201300455
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发表时间：
2013-04-01
影响因子：
4.3
通讯作者：
Guo, Wei
中科院分区：
文献类型：
--
作者：
Liu, Jing;Sun, Yuan-Qiang;Guo, Wei
传统上，硫化氢（H2S）被认为是有毒气体。后来有报道称血液中H2S的典型浓度在10-100 μm范围内，提示其与生物功能有关。事实上，最近的研究认为H2S是除一氧化氮（NO）和一氧化碳（CO）之外的第三种气体传递物例如，H2S已被认为可以介导广泛的生理效应，包括调节细胞生长、血管舒张和血管生成、调节神经传递、抑制胰岛素信号和调节炎症此外，它还可以作为一种抗氧化剂或活性氧（ROS）的清除剂此外，研究表明，它的放松与阿尔茨海默病、唐氏综合症、糖尿病、肝硬化的症状有关虽然H2S已被认为与各种生理和病理功能有关，但其许多潜在的分子事件仍不为人所知。因此，迫切需要有效的方法来敏感和选择性地检测生命系统中的H2S。荧光光谱由于其简单、灵敏、实时成像，特别是其对活细胞或组织中目标生物分子的无损检测，是痕量样品传感和成像的有力工具。毫无疑问，我们对细胞生物学中分子功能的理解大大受益于荧光探针的进步。一般来说，设计用于检测生命系统中H2S的荧光探针涉及几个重大挑战：一是获得对其他生物硫醇的足够选择性，包括还原型谷胱甘肽（GSH，存在于约1-10 mm的水平）和半胱氨酸（l-Cys，约100 μm）；二是要达到足够的灵敏度，因为据报道，生理学上相关的H2S浓度范围从纳米到毫摩尔不等；第三个挑战是在温和条件下的快速反应，因为H2S代谢迅速。以前报道的硫化氢荧光探针主要集中在体外检测系统近年来，有可能用于生命系统中H2S检测的荧光探针已经出现，[7]和相应的设计策略是基于H2S的几个重要特性，即双亲核性，[8-10]对叠氮化物、硝基和羟基- achtungtrennungamine基团的良好还原性，[11-13]对铜离子的高结合亲和力，[14]和对二硝基苯醚的高效硫解这些独特的性质可以有效地将H2S与其他互补物质区分，特别是在大多数细胞内发现的毫摩尔浓度的生物硫醇，从而可以在复杂的生物系统中检测H2S。然而，除了Wang和Nagano分别报道的叠氮丹酰[12a]和hsp -1 [14a]（在几秒内）外，大多数报道的H2S探针显示延迟响应时间（超过20min），因此，对于实时测定生物系统中H2S浓度的波动并不满意。这是一个有待解决的问题。比率荧光探针可以消除干扰信号输出的大部分或全部因素，如仪器效率、环境条件（pH、极性、温度等）和探针的定位，通过内置两个发射波段的校正，因此，与基于荧光强度的探针相比，比率荧光探针更有利虽然已经报道了一些硫化氢的比例荧光探针，[10,12d, 17]，但它们中的大多数仍然显示出延迟的响应时间(超过20…
Traditionally, hydrogen sulfide (H2S) was considered to be a toxic gas. Later, it was reported that the typical concentration of H2S in blood is in the range 10–100 μm,[1] suggesting its connection with biological functions. Indeed, recent studies regard H2S as the third gaseous transmitter besides nitric oxide (NO) and carbon monoxide (CO).[2] For example, H2S has been recognized to mediate a wide range of physiological effects, including regulation of cell growth, vasodilation, and angiogenesis, mediation of neurotransmission, inhibition of insulin signaling, and regulation of inflammation.[3] Moreover, it acts as an antioxidant or scavenger of reactive oxygen species (ROS).[4] Furthermore, studies have shown that its deregulation is correlated with the symptoms of Alzheimer s disease, Down s syndrome, diabetes, and liver cirrhosis.[5] Although H2S has been recognized to be linked to various physiological and pathological functions, many of its underlying molecular events remain unknown. Therefore, efficient methods to sensitively and selectively detect H2S in living systems are urgently required. Fluorescence spectroscopy is a powerful tool for sensing and imaging trace amounts of samples because of its simplicity, sensitivity, real-time imaging, and especially its nondestructive detection of target biomolecules in live cells or tissues. Undoubtedly, our understanding of molecular function in cell biology has significantly benefited from advancements of fluorescent probes. Generally, the design of fluorescent probes for detection of H2S in living systems involves several substantial challenges: one is to attain sufficient selectivity over other biothiols, including reduced glutathione (GSH, present at levels of about 1–10 mm) and lcysteine (l-Cys, about 100 μm); the other is to achieve sufficient sensitivity, because the physiologically relevant H2S concentration has been reported to range from nano-to millimolar levels; and the third challenge is fast response under mild condition, because H2S is metabolized rapidly. Previously reported fluorescent probes for H2S are mainly focused on in vitro assay systems.[6] Recently, fluorescent probes that can potentially be used for H2S detection in living systems have become available,[7] and the corresponding design strategies are based on several significant characteristic properties of H2S, namely dual nucleophilicity,[8–10] good reducing property towards azide, nitro and hydroxy-ACHTUNGTRENNUNGamine groups,[11–13] high binding affinity towards copper ion,[14] and efficient thiolysis of dinitrophenyl ether.[15] These unique properties can efficiently differentiate H2S from other completive species, in particular, millimolar concentrations of biothiols found inside most cells, thereby enabling the detection of H2S in complicated biological systems. However, most of the reported H2S probes display a delayed response time (more than 20min)—except for dansyl azide [12a] and HSip-1 [14a](within seconds) reported by Wang and Nagano, respectively—and, thus, are not satisfactory for real-time determination of the fluctuations in H2S concentration in biological systems. This is a problem to be solved. Ratiometric fluorescent probes can eliminate most or all the factors that interfere with the signal output, such as instrumental efficiency, environmental conditions (pH, polarity, temperature, and so forth), and the localization of the probe, by built-in correction of two emission bands, and, thus, are more favorable in comparison with fluorescence intensity-based probes.[16] Although a few ratiometric fluorescent probes for H2S have been reported,[10, 12d, 17] most of them still display a delayed response time (more than 20 …