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目的 研究重组人组织型纤溶酶原激活剂缺失变体(reteplase,Ret)的溶栓作用以及对离体家兔血小板聚集功能和对血小板静息状态胞浆内游离钙浓度的影响。方法 家兔血栓模型采用动-静脉旁路法,血小板聚集实验采用比浊法,血小板静息状态胞浆内游离钙浓度采用Fura-2荧光测定法。结果Ret(3.75×105,7.50× 105,15.0×105U·kg-1)对家兔动-静脉旁路所形成的血栓有明显溶栓作用,可明显减轻血栓干湿重量,并呈一定的剂量依赖性。Ret(1 000,3 000,10 000 U·mL-1)对ADP诱导的血小板聚集可产生浓度依赖性的抑制作用,并可明显降低血小板静息状态的[Ca2+]i,并呈一定的浓度依赖性。结论Ret具有明显的血栓溶解作用,可明显抑制ADP诱导的血小板聚集,并明显降低血小板静息状态的[Ca2+]i。
Objective To investigate the thrombolytic effect of recombinant human tissue plasminogen activator (RetA) and its effect on platelet aggregation and intracytoplasmic free calcium concentration of resting platelets in vitro. Methods Rabbit thrombosis model was established by arterio-venous bypass method. Platelet aggregation was assayed by turbidimetry and cytosolic free calcium concentration at platelet resting state was measured by Fura-2 fluorescence assay. Results Ret (3.75 × 105,7.50 × 105,15.0 × 105U · kg-1) had significant thrombolytic effect on thrombus formed by arterial-venous bypass in rabbits, which could significantly reduce the weight of thrombus, Dependency. Ret (1 000, 3 000, 10 000 U · mL-1) produced a concentration-dependent inhibition of platelet aggregation induced by ADP, and significantly reduced platelet resting [Ca2 +] i, Dependency. Conclusion Ret has obvious thrombolytic effect, which can significantly inhibit platelet aggregation induced by ADP and significantly reduce platelet resting state [Ca2 +] i.