Inhibition of arterial thrombosis by recombinant annexin V in a rabbit carotid artery injury model.
Inhibition of arterial thrombosis by recombinant annexin V in a rabbit carotid artery injury model.
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DOI：
10.1161/01.cir.96.7.2339
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发表时间：
1997-10
期刊：
影响因子：
37.8
通讯作者：
P. Thiagarajan;C. Benedict
中科院分区：
文献类型：
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作者：
P. Thiagarajan;C. Benedict
背景技术阴离子磷脂的促凝血作用可能在动脉血栓形成的发展中起主要作用。方法和结果膜联蛋白V是一种钙依赖性阴离子磷脂结合蛋白，从大肠杆菌中表达和分离，并在兔颈动脉血栓形成模型中检测其抗血栓作用。通过施加电流产生大约50%的管腔闭塞，在左颈动脉中形成部分闭塞的血栓。电流停止后，由于血小板/纤维蛋白血栓持续形成，流动在 42+/-12 分钟内完全停止（n=6）。当从电流停止时开始，以2.8至16.6微克×千克(-1)×分钟(-1)的剂量给予膜联蛋白V 180分钟时，对血栓形成产生剂量依赖性抑制。在剂量为5.6μg×kg(-1)×min(-1)时，血流在整个输注过程中以及输注停止后的另外60分钟内保持畅通。此外，血栓重量减少（16+/-7.4 对比 2.0+/-1.0 g）、(125)I-纤维蛋白沉积（减少约 45%，P<.001）和 (111)In 标记的血小板积累（减少约 43%，P<.001）。预先将膜联蛋白V与磷脂酰丝氨酸胶束混合消除了膜联蛋白V的抗血栓作用，而与磷脂酰胆碱胶束混合则没有效果。膜联蛋白 V 的抗血栓作用与出血倾向无关，这是通过放置在延伸至肌肉组织的手术切口中的纱布垫中吸收的血液量和标准模板出血时间来判断的。结论 这些观察结果支持血小板中阴离子磷脂暴露在动脉血栓形成中的潜在重要作用，并且抑制这种活性可能成为冠状动脉血栓形成和中风以及血管成形术后治疗的新靶点。
BACKGROUND The procoagulant effect of anionic phospholipid may play a major role in the development of arterial thrombosis. METHODS AND RESULTS Annexin V, a calcium-dependent anionic-phospholipid-binding protein, was expressed and isolated from Escherichia coli and its antithrombotic effect examined in a rabbit carotid artery thrombosis model. A partially occlusive thrombus was formed in the left carotid artery by application of electric current to produce an approximately 50% occlusion of the lumen. After the current was discontinued, flow ceased completely within 42+/-12 minutes (n=6) because of continuing platelet/fibrin thrombus formation. When annexin V was given at doses of 2.8 to 16.6 microg x kg(-1) x min(-1) for a period of 180 minutes, starting at the time the current was stopped, there was a dose-dependent inhibition of thrombus formation. At a dose of 5.6 microg x kg(-1) x min(-1), blood flow remained patent throughout the infusion and for an additional 60 minutes after the infusion was stopped. In addition, there was a decrease in thrombus weight (16+/-7.4 versus 2.0+/-1.0 g), (125)I-fibrin deposition (approximately 45% reduction, P<.001), and (111)In-labeled platelet accumulation (approximately 43% reduction, P<.001). Prior mixing of annexin V with phosphatidylserine micelles abolished the antithrombotic effect of annexin V, whereas mixing with phosphatidylcholine micelles had no effect. The antithrombotic effect of annexin V was not associated with bleeding tendency, as judged by the amount of blood absorbed in a gauze pad placed in a surgical incision extending to the muscle tissue and by the standard template bleeding time. CONCLUSIONS These observations support a potentially important role for anionic phospholipid exposure in platelets in arterial thrombosis, and inhibition of this activity could be a novel target for therapy in coronary thrombosis and stroke and after angioplasty.