Zaman A G, Osende J I, Chesebro J H, Fuster V, Padurean A, Gallo R, Worthley S G, Helft G, Rodriguez O X, Fallon J T, Badimon J J
Cardiovascular Biology Research Laboratory, Mount Sinai School of Medicine, New York, NY 10029-6574, USA.
Arterioscler Thromb Vasc Biol. 2000 Mar;20(3):860-5. doi: 10.1161/01.atv.20.3.860.
Current methods for monitoring thrombosis and thrombus growth are invasive and provide only single-time-point data. Animal models rely mainly on flow changes as a surrogate of thrombus formation. Our aim was to validate a unique potentially noninvasive system to detect and quantify dynamic thrombus formation in vivo by using a porcine model of carotid artery injury. Thrombus growth was monitored by deposition of autologous (111)In-labeled platelet activity over the injured artery by use of miniaturized gamma detectors and Doppler blood flow. Counts were recorded at 2-minute intervals for 2 hours. The technique was validated by comparing standard antithrombotic agents against controls. Platelet recruitment was detected before significant change in flow. Thrombus formation, calculated as the area under the curve (platelets x minutes x 10(6)), was greatest for control animals (11.7+/-1.28), followed by animals treated with aspirin (6.13+/-0.91, P<0.05), heparin (2.45+/-0.34, P<0.05), and hirudin (0.2+/-0.01, P<0.01 compared with heparin). The rate of platelet deposition was assessed as the slope of the curve in the first 30 minutes (platelets x 10(6) per minute) for the following treatment groups of animals: control, 3.53+/-0.34; aspirin, 1.67+/-0. 34 (P<0.01); heparin, 1.55+/-0.3 (P<0.01); and hirudin, 0.25+/-0.03 (P<0.001). There was no statistical difference between heparin and aspirin treatments. Change in flow was assessed as reduction from baseline: control, >99+/-0.34%; aspirin, 39+/-9.1%; heparin, 36+/-12. 5%; and hirudin, 17+/-5.4%. There was no statistical difference between the aspirin- and heparin-treated groups. Morphometric analysis revealed >99+/-0.63% occlusion of the luminal area with thrombus for the control group, 43+/-14.3% for the aspirin-treated group, 30+/-5.6% for the heparin-treated group, and <10+/-1.8% for the hirudin-treated group. Assessment of platelet-thrombus formation with this technique was more sensitive than change in flow in determining antithrombotic efficacy, and thrombus formation was detected earlier. This study validates a new quantitative, sensitive, potentially noninvasive, portable, in vivo monitoring of dynamic thrombus growth, which appears applicable to phase II studies in humans.
目前用于监测血栓形成和血栓生长的方法具有侵入性，且仅能提供单次时间点的数据。动物模型主要依赖血流变化作为血栓形成的替代指标。我们的目的是通过使用颈动脉损伤的猪模型，验证一种独特的、潜在非侵入性的系统，用于在体内检测和量化动态血栓形成。通过使用小型伽马探测器和多普勒血流仪，将自体（111）铟标记的血小板活性沉积在受损动脉上，监测血栓生长。每隔2分钟记录一次计数，持续2小时。通过将标准抗血栓药物与对照组进行比较，验证了该技术。在血流出现显著变化之前就检测到了血小板募集。以曲线下面积（血小板×分钟×10⁶）计算的血栓形成，对照组动物最大（11.7±1.28），其次是阿司匹林治疗组（6.13±0.91，P<0.05）、肝素治疗组（2.45±0.34，P<0.05）和水蛭素治疗组（与肝素相比，0.2±0.01，P<0.01）。评估以下治疗组动物在最初30分钟内曲线的斜率（每分钟血小板×10⁶）作为血小板沉积率：对照组，3.53±0.34；阿司匹林组，1.67±0.34（P<0.01）；肝素组，1.55±0.3（P<0.01）；水蛭素组，0.25±0.03（P<0.001）。肝素和阿司匹林治疗之间无统计学差异。将血流变化评估为相对于基线的降低：对照组，>99±0.34%；阿司匹林组，39±9.1%；肝素组，36±12.5%；水蛭素组，17±5.4%。阿司匹林和肝素治疗组之间无统计学差异。形态计量学分析显示，对照组管腔面积被血栓阻塞>99±0.63%，阿司匹林治疗组为43±14.3%，肝素治疗组为30±5.6%，水蛭素治疗组<10±1.8%。用该技术评估血小板血栓形成在确定抗血栓疗效方面比血流变化更敏感，且血栓形成检测得更早。本研究验证了一种新的定量、敏感、潜在非侵入性、便携式的体内动态血栓生长监测方法，该方法似乎适用于人类的II期研究。
