In vivo Imaging of thrombin activity in experimental thrombi with thrombin-sensitive near-infrared molecular probe
In vivo Imaging of thrombin activity in experimental thrombi with thrombin-sensitive near-infrared molecular probe
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DOI：
10.1161/01.atv.0000033089.56970.2d
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发表时间：
2002-11-01
影响因子：
8.7
通讯作者：
Weissleder, R
中科院分区：
文献类型：
--
作者：
Jaffer, FA;Tung, CH;Weissleder, R
目的-凝血酶是一种丝氨酸蛋白酶，在血栓形成以及其他细胞和发育过程中发挥重要作用。在这项研究中，我们研究了一种新型的凝血酶激活的分子探针提供实验性血栓中凝血酶活性的在体图像的能力。方法和结果-凝血酶探针由一种近红外(NIR)荧光色素通过凝血酶特异性寡肽底物连接到运送载体上。在人类血液中，内源性凝血酶激活了凝血酶探针，使荧光信号增加了18倍(P=0.008)。特异性凝血酶抑制剂水飞蓟素可抑制探针激活82%(P=0.007)。然后在急性实验性小鼠血栓形成模型中研究体内凝血酶活性的成像，最长可达12小时。全身注射凝血酶探针后，在急性和亚急性血栓内可迅速检测到局部近红外荧光信号增强。相反，在与对照的近红外荧光染料类似的实验中，没有发现血栓信号增强。结论：通过使用一种新型的凝血酶可激活和凝血酶特异性的近红外分子探针，可以在体内成像凝血酶活性。凝血酶探针可以加深对凝血酶在血栓形成和其他动态平衡和病理条件下的作用的了解。
Objective-Thrombin, a serine protease, plays an important role in thrombosis as well as other cellular and developmental processes. In this study, we investigated the ability of a novel thrombin-activatable molecular probe to provide in vivo images of thrombin activity in experimental thrombi.Methods and Results-The thrombin probe consists of a near-infrared (NIR) fluorochrome attached to a delivery vehicle via a thrombin-specific oligopeptide substrate. In human blood, endogenous thrombin activated the thrombin probe and increased the fluorescence signal by 18-fold (P = 0.008). Hirudin, a specific thrombin inhibitor, suppressed probe activation by 82% (P = 0.007). Imaging of in vivo thrombin activity was then investigated in acute experimental murine thrombosis models up to 12 hours. After systemic thrombin probe injection, focal NIR fluorescence signal enhancement was rapidly detected within acute and subacute thrombi. In contrast, no thrombosis signal enhancement was seen in similar experiments with a control NIR fluorochrome,Conclusions-Thrombin activity can be imaged in vivo by using a novel thrombin-activatable and thrombin-specific NIR molecular probe. The thrombin probe could enhance the understanding of the role of thrombin in thrombogenesis and other homeostatic and pathological conditions.