A Novel Pentapeptide Targeting Integrin β3-Subunit Inhibits Platelet Aggregation and Its Application in Rat for Thrombosis Prevention.
A Novel Pentapeptide Targeting Integrin β3-Subunit Inhibits Platelet Aggregation and Its Application in Rat for Thrombosis Prevention.
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DOI：
10.3389/fphar.2016.00049
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发表时间：
2016
影响因子：
5.6
通讯作者：
Wu G
中科院分区：
文献类型：
--
作者：
Qu Q;Liu Y;Yan X;Fan X;Liu N;Wu G
背景：抗血小板治疗在血栓性疾病的预防和治疗中起着关键作用。我们报道了从结缔组织生长因子c端筛选P1C作为一种新的整合素结合肽。初步研究表明，P1C具有抑制血小板聚集的潜力。目的：寻找P1C片段中最短的活性单元，探讨其体内和体外活性。方法：制备一系列截断的P1C片段并进行抗血小板活性筛选。最活跃的片段用凝血试验评估。利用流式细胞术和共聚焦显微镜检测肽与整合素之间的相互作用。利用两种大鼠模型进一步探讨其体内电位。结果：从一系列截断的P1C形式中，筛选出5个氨基酸的所谓P1Cm肽，即IRTPK，是最短的活性单位，具有较好的活性。凝血实验和体内毒性实验表明，P1Cm在体内是安全的，并且在体外以浓度依赖的方式抑制ADP和th诱导的人血小板聚集。此外，它对混凝参数的影响有限。流式细胞术和共聚焦显微镜实验一致表明，肽特异性结合血小板上整合素的β3亚基。进一步通过大鼠动静脉分流和颈动脉血栓形成模型实验表明，P1Cm能有效预防血栓形成。结论：P1Cm可能是一种新的，有前途的抗血栓治疗替代目前可用的抗血小板治疗。
Background: Antiplatelet therapy plays a pivotal role in the prevention and treatment of thrombotic diseases. We reported the screening of P1C as a novel integrin-binding peptide from the C-terminal of connective tissue growth factor. Primary study indicated that P1C has potential against platelet aggregation. Objectives: We aimed to find the shortest active unit from the P1C fragments and explore its in vivo and in vitro activities. Methods: A series of truncated P1C fragments was prepared and screened for antiplatelet activity. The most active fragment was evaluated using coagulation assays. Flow cytometry and confocal microscopy were used to determine the interaction between the peptide and the integrin. The in vivo potential was further explored using two types of rat models. Results: From a series of truncated P1C forms, a so-called P1Cm peptide of 5-amino acids, namely, IRTPK was screened out as the shortest active unit with superior activity. Coagulation experiments and an in vivo toxicity assay demonstrated that P1Cm is safe in vivo and inhibits ADP- and TH-induced human platelet aggregation in vitro in a concentration-dependent manner. Furthermore, it has limited effect on the coagulation parameters. Flow cytometry and confocal microscopy experiments consistently indicated that the peptide specifically binds the β3-subunit of integrin on platelets. Further experiments using rat models of artery-vein shunt and carotid arterial thrombosis illustrated that P1Cm can effectively prevent thrombosis formation. Conclusion: P1Cm may be a new, promising antithrombotic alternative to currently available antiplatelet treatments.