槐定碱下调PI3K/AKT抑制血小板整合素αIIbβ3活化抗血栓机制研究

血栓性疾病严重危害人类健康，抗血小板药物的开发对于该类疾病的治疗意义重大。槐定碱是从中药苦豆子中提取的单体生物碱，已作为抗肿瘤药物上市。我们前期研究发现：槐定碱抑制体外血栓形成，抑制多种激动剂诱导的血小板聚集。并且，槐定碱抑制血小板聚集和血栓形成的关键节点事件——整合素αIIbβ3的活化，下调参与介导整合素αIIbβ3活化的PI3K/AKT。因此，我们提出假设：槐定碱通过下调PI3K/AKT抑制血小板整合素αIIbβ3活化发挥抗血栓功效。本项目拟采用多种体内血栓模型，证实槐定碱的体内抗血栓功效；从细胞和分子水平证实槐定碱通过下调PI3K/AKT抑制整合素αIIbβ3活化；通过分子生物学手段进一步阐明槐定碱对PI3K/AKT的抑制作用；设计光亲和探针，结合质谱技术，鉴定槐定碱下调PI3K/AKT的靶标蛋白质。本项目的完成，将阐明槐定碱的抗血栓机制，为槐定碱的临床应用提供新的理论依据。

Thrombotic diseases seriously threaten human health, and the development of antiplatelet drugs is of great significance for the treatment of such diseases. Sophoridine is a monomeric alkaloid extracted from the traditional Chinese medicine Sophora alopecuroides L., and it is marketed as an anti-tumor drug. Our previous research found that, sophoridine inhibits thrombus formation in vitro, and inhibits platelet aggregation induced by various agonists. Moreover, sophoridine inhibits the activation of integrin αIIbβ3, a key node event of platelet aggregation and thrombosis, and down-regulates PI3K/AKT, which is involved in mediating the activation of integrin αIIbβ3. Therefore, we propose the hypothesis that, sophoridine can inhibit the activation of platelet integrin αIIbβ3 by down-regulating PI3K/AKT to exert its antithrombotic effect. This project will use a variety of in vivo thrombosis models to confirm the antithrombotic effect of sophoridine in vivo; confirm that sophoridine inhibits the activation of integrin αIIbβ3 through down-regulating PI3K/AKT in the cellular and molecular levels; further clarify the Inhibition of PI3K/AKT sophoridine through molecular biological methods; identify the target protein of sophoridine down-regulating PI3K/AKT by designing a photo-affinity probe and mass spectrometry technology. This study will clarify the antithrombotic mechanism of sophoridine, and provide a new theoretical basis for the clinical application of sophoridine.