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首页> 外文期刊>Canadian Journal of Physiology and Pharmacology >Evaluation of a multifunctional staphylokinase variant with thrombin inhibition and antiplatelet aggregation activities produced from salt-inducible E. coli GJ1158
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Evaluation of a multifunctional staphylokinase variant with thrombin inhibition and antiplatelet aggregation activities produced from salt-inducible E. coli GJ1158

机译：评估盐诱导的大肠杆菌GJ1158产生的具有凝血酶抑制和抗血小板聚集活性的多功能葡萄激酶激酶变体

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摘要

Reocclusion is one of the major root causes for secondary complications that arise during thrombolytic therapy. A multifunctional staphylokinase variant SRH (staphylokinase (SAK) linked with tripeptide RGD and didecapeptide Hirulog) with antiplatelet and antithrombin activities in addition to clot specific thrombolytic function, was developed to address the reocclusion problem. We preferred to use Escherichia coli GJ1158 as the host in this study for economic production of SRH by osmotic (0.3 mol/L sodium chloride) induction, to overcome the problems associated with the yeast expression system. The therapeutic potential of SRH was evaluated in the murine model of vascular thrombosis. The SAK protein (1 mg/kg body mass) and SRH protein (1 mg/kg and 2 mg/kg) were administered intravenously to the different treatment groups. The results have shown a dose-dependent antithrombotic effect in carrageenan-induced mouse tail thrombosis. The thrombin time, activated partial thromboplastin time, and prothrombin time were significantly prolonged (p < 0.05) in the SRH-infused groups. Moreover, SRH inhibited platelet aggregation in a dose-dependent manner (p < 0.05), while the bleeding time was significantly (p < 0.05) prolonged. All of these results inferred that the osmotically produced multifunctional fusion protein SRH (SAK-RGD-Hirulog) is a promising thrombolytic agent, and one which sustained its multifunctionality in the animal models.
机译：再闭塞是溶栓治疗过程中引起继发性并发症的主要原因之一。为了解决再阻塞问题，开发了一种具有抗血小板和抗凝血酶活性的多功能葡萄球菌激酶变体SRH（与三肽RGD和二肽肽Hirulog连接的葡萄球菌酶（SAK））。在本研究中，我们优选使用大肠杆菌GJ1158作为宿主，通过渗透（0.3 mol / L氯化钠）诱导经济生产SRH，以克服与酵母表达系统相关的问题。在鼠的血管血栓形成模型中评估了SRH的治疗潜力。将SAK蛋白（1 mg / kg体重）和SRH蛋白（1 mg / kg和2 mg / kg）静脉内给药至不同的治疗组。结果显示了角叉菜胶诱导的小鼠尾巴血栓形成的剂量依赖性抗血栓形成作用。在注入SRH的组中，凝血酶时间，活化的部分凝血活酶时间和凝血酶原时间显着延长（p <0.05）。此外，SRH以剂量依赖的方式抑制血小板凝集（p <0.05），而出血时间明显延长（p <0.05）。所有这些结果推断，渗透产生的多功能融合蛋白SRH（SAK-RGD-Hirulog）是一种有前途的溶栓剂，并且在动物模型中可以维持其多功能性。

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