Department of Medicine, Center for Metabolic Disease Research, Tokai University School of Medicine, Isehara, Japan.
J Atheroscler Thromb. 2009;16(6):807-14. doi: 10.5551/jat.2030. Epub 2009 Dec 22.
We investigated thrombus formation at the site of functional injury to endothelial cells by FeCl(3).
After preparation of cremasteric arteries of mice, controlled endothelial injury was induced by application of FeCl(3). Endothelial cells were rendered fluorescent by addition of FITC (fluorescein isothiocyanate)-labeled isolectin B4. Circulating platelets and leukocytes were made fluorescent by rhodamine 6G. Three-dimensional (3D) growth of thrombi was visualized in real time. Effects of aspirin and clopidogrel pre-treatments on the growth of thrombi were investigated in vivo as well as in an ex vivo flow chamber system.
Endothelial cells were tightly bound to each other to protect local thrombus formation. Platelets started to adhere to endothelial cells when FeCl(3) was applied. Three-dimensional growth of thrombi, which takes 10.6+/-7.5 minutes for complete occlusion in control, can be visualized with our imaging system. Aspirin pre-treatment at the dose tested did not influence either endothelial injury or platelet thrombus growth, while clopidogrel pretreatment significantly inhibited 3D growth and prolonged occlusion time up to 64.6+/-25.3 minutes (100 mg/kg). A similar inhibiting effect of clopidogrel was reproduced in ex vivo flow chamber experiments.
We have developed an in vivo system to detect thrombus formation after functional damage to the endothelium.
我们研究了 FeCl(3) 引起的内皮细胞功能损伤部位血栓形成的情况。
在制备小鼠提睾肌动脉后，通过应用 FeCl(3) 诱导内皮细胞损伤。通过添加 FITC（异硫氰酸荧光素）标记的 B4 型大豆凝集素使内皮细胞发荧光。通过 rhodamine 6G 使循环血小板和白细胞发荧光。实时可视化血栓的三维（3D）生长。体内和体外流动腔系统研究了阿司匹林和氯吡格雷预处理对血栓生长的影响。
内皮细胞紧密结合在一起，以保护局部血栓形成。当应用 FeCl(3) 时，血小板开始黏附在内皮细胞上。我们的成像系统可以可视化血栓的 3D 生长，在对照中完全闭塞需要 10.6+/-7.5 分钟。在测试剂量下，阿司匹林预处理既不影响内皮细胞损伤，也不影响血小板血栓生长，而氯吡格雷预处理显著抑制 3D 生长，并将闭塞时间延长至 64.6+/-25.3 分钟（100mg/kg）。在体外流动腔实验中也重现了氯吡格雷的类似抑制作用。
我们已经开发出一种体内系统，用于检测内皮细胞功能损伤后血栓形成。
