DataSheet1_Matrine Impairs Platelet Function and Thrombosis and Inhibits ROS Production.ZIP

Matrine is a naturally occurring alkaloid and possesses a wide range of pharmacological properties, such as anti-cancer, anti-oxidant, anti-inflammatory effects. However, whether it affects platelet function and thrombosis remains unclear. This study aims to evaluate the effect of matrine on platelet function and thrombus formation. Human platelets were treated with matrine (0–1 mg/ml) for 1 h at 37°C followed by measuring platelet aggregation, granule secretion, receptor expression by flow cytometry, spreading and clot retraction. In addition, matrine (10 mg/kg) was injected intraperitoneally into mice to measure tail bleeding time, arterial and venous thrombus formation. Matrine dose-dependently inhibited platelet aggregation and ATP release in response to either collagen-related peptide (Collagen-related peptide, 0.1 μg/ml) or thrombin (0.04 U/mL) stimulation without altering the expression of P-selectin, glycoprotein Ibα, GPVI, or αIIbβ3. In addition, matrine-treated platelets presented significantly decreased spreading on fibrinogen or collagen and clot retraction along with reduced phosphorylation of c-Src. Moreover, matrine administration significantly impaired the in vivo hemostatic function of platelets, arterial and venous thrombus formation. Furthermore, in platelets stimulated with CRP or thrombin, matrine significantly reduced Reactive oxygen species generation, inhibited the phosphorylation level of ERK1/2 (Thr202/Tyr204), p38 (Thr180/Tyr182) and AKT (Thr308/Ser473) as well as increased VASP phosphorylation (Ser239) and intracellular cGMP level. In conclusion, matrine inhibits platelet function, arterial and venous thrombosis, possibly involving inhibition of ROS generation, suggesting that matrine might be used as an antiplatelet agent for treating thrombotic or cardiovascular diseases.

马蹄碱是一种天然存在的生物碱，具备广泛的药理特性，诸如抗癌、抗氧化、抗炎作用。然而，其是否影响血小板功能及血栓形成尚不明确。本研究旨在评估马蹄碱对血小板功能及血栓形成的影响。将人血小板在37°C下用马蹄碱（0–1 mg/ml）处理1小时，随后通过流式细胞术检测血小板聚集、颗粒分泌、受体表达、扩散和凝血块收缩。此外，将马蹄碱（10 mg/kg）腹腔注射至小鼠体内，以测量尾出血时间、动脉和静脉血栓形成。结果显示，马蹄碱剂量依赖性地抑制了血小板在胶原蛋白相关肽（Collagen-related peptide，0.1 μg/ml）或凝血酶（0.04 U/mL）刺激下的聚集和ATP释放，而不改变P-选择素、糖蛋白Ibα、GPVI或αIIbβ3的表达。此外，马蹄碱处理后的血小板在纤维蛋白原或胶原蛋白上的扩散以及凝血块收缩显著降低，伴随c-Src磷酸化水平的减少。再者，马蹄碱给药显著损害了血小板在体内的止血功能，以及动脉和静脉血栓的形成。此外，在CRP或凝血酶刺激的血小板中，马蹄碱显著降低了活性氧的产生，抑制了ERK1/2（Thr202/Tyr204）、p38（Thr180/Tyr182）和AKT（Thr308/Ser473）的磷酸化水平，并增加了VASP磷酸化（Ser239）和细胞内cGMP水平。综上所述，马蹄碱通过抑制活性氧的产生来抑制血小板功能、动脉和静脉血栓形成，可能涉及对ROS生成的抑制，表明马蹄碱可能被用作抗血小板药物，以治疗血栓或心血管疾病。