Yu Xiao-Qin, Zhang Qi-Yun, Zhou Shu-Ting, Lu Qing-Yu, Sun Qian-Yun
State Key Laboratory of Discovery and Utilization of Functional Components in Traditional Chinese Medicine, Guizhou Medical University, Guiyang, China.
School of Pharmaceutical Sciences, Guizhou Medical University, Guiyang, China.
J Venom Anim Toxins Incl Trop Dis. 2025 Jul 4;31:e20250003. doi: 10.1590/1678-9199-JVATITD-2025-0003. eCollection 2025.
Snake venom C-type lectin-like proteins (also known as snaclecs) have anticoagulation and procoagulation effects by targeting platelet or coagulation factor IX/X, suggesting their potential as candidates for new anticoagulant drugs. Therefore, this study aims to evaluate the antiplatelet and antithrombotic effects of a new snaclec from venom and its potential as an anticoagulant candidate.
Promucetin was purified through sequential column chromatography, and its molecular mass was determined by SDS-PAGE. The α- and β-chains of promucetin were identified using liquid chromatography-mass spectrometry (LC-MS). analyses of platelet aggregation were performed using turbidimetric methods, thromboelastography, and coagulation activity assays. For experiments, promucetin was administered to rats at varying concentrations, and platelet changes were monitored. The antithrombotic effects of promucetin were assessed using a FeCl₃-induced rat thrombosis model.
Promucetin existed as two multimers with molecular weights of 140.1 kDa and 91.9 kDa under non-reducing conditions. Sequence analysis revealed that its α-chain and β-chain shared 71% and 34% homology, respectively, with TMVA from the same snake venom. platelet aggregation assays indicated that promucetin activated platelets via glycoprotein Ib. Thromboelastography showed that promucetin inhibited both coagulation factor activity and platelet function, resulting in an anticoagulant effect. Specifically, thrombin time was prolonged, while activated partial thromboplastin time and prothrombin time remained unchanged. , promucetin administration led to a dose-dependent decrease in platelet count. At doses of 25 and 50 μg/kg, promucetin significantly inhibited thrombosis, with inhibition rates of 40.9% and 74.4%, respectively. For comparison, lysine acetylsalicylate produced an inhibition rate of 36.7%.
Promucetin exhibits significant ability to modulate coagulation function and effectively inhibit thrombosis by activating platelet via GPIb and reducing platelet count, which helps us understand its biological function in snake bites, it exhibits the potential to be a candidate for anticoagulant therapy.
蛇毒C型凝集素样蛋白（也称为蛇凝集素）通过靶向血小板或凝血因子IX/X发挥抗凝和促凝作用，表明它们有潜力成为新型抗凝药物的候选物。因此，本研究旨在评估一种来自蛇毒的新型蛇凝集素的抗血小板和抗血栓形成作用及其作为抗凝候选物的潜力。
通过连续柱色谱法纯化前粘蛋白，并用SDS-PAGE测定其分子量。使用液相色谱-质谱联用仪（LC-MS）鉴定前粘蛋白的α链和β链。采用比浊法、血栓弹力图法和凝血活性测定法进行血小板聚集分析。在实验中，将不同浓度的前粘蛋白给予大鼠，并监测血小板变化。使用FeCl₃诱导的大鼠血栓形成模型评估前粘蛋白的抗血栓形成作用。
在前粘蛋白在非还原条件下以两种分子量分别为140.1 kDa和91.9 kDa的多聚体形式存在。序列分析表明，其α链和β链与来自同一种蛇毒的TMVA的同源性分别为71%和34%。血小板聚集试验表明，前粘蛋白通过糖蛋白Ib激活血小板。血栓弹力图显示，前粘蛋白抑制凝血因子活性和血小板功能，从而产生抗凝作用。具体而言，凝血酶时间延长，而活化部分凝血活酶时间和凝血酶原时间保持不变。此外，给予前粘蛋白导致血小板计数呈剂量依赖性下降。在25和50 μg/kg的剂量下，前粘蛋白显著抑制血栓形成，抑制率分别为40.9%和74.4%。相比之下，赖氨酸乙酰水杨酸酯的抑制率为36.7%。
前粘蛋白具有显著调节凝血功能的能力，并通过GPIb激活血小板和减少血小板计数有效抑制血栓形成，这有助于我们了解其在蛇咬伤中的生物学功能，它有潜力成为抗凝治疗的候选物。
