Xanthium strumarium L. Exhibits Potent Antiplatelet and Antithrombotic Effects by Modulating MAPK and PI3K/AKT Signaling Pathways and Inhibiting Ferric Chloride-Induced Thrombosis

<b>Background:</b> Cardiovascular diseases, driven by platelet hyperactivation and thrombosis, remain the leading global cause of death. Excessive platelet activation contributes to atherosclerosis and thrombo-inflammatory disorders, underscoring the urgent need for safer and more effective antiplatelet agents. <b>Objectives:</b><i>Xanthium strumarium</i> L. (<i>X. strumarium</i>) has been reported to exhibit a wide range of pharmacological effects, including anti-inflammatory and antioxidant activities. However, its antiplatelet and antithrombotic effects remain unexplored. Therefore, the present study aimed to comprehensively evaluate the antiplatelet and antithrombotic effects of <i>X. strumarium</i> through integrated in vitro and in vivo experiments. <b>Methods:</b> The principal bioactive compounds present in the <i>X. strumarium</i> extract were identified through GC-MS analysis. In vitro antiplatelet effects were evaluated via light transmission aggregometry, scanning electron microscopy (SEM), ATP and calcium mobilization assays, αIIbβ3 binding assay, clot retraction assay, and Western blotting. In vivo ferric chloride-induced (FeCl₃) murine thrombus model was established to evaluate thrombogenesis. <b>Results:</b> Our results demonstrated that <i>X. strumarium</i> at 25, 50, or 100 μg/mL significantly inhibited collagen, ADP, U46619, and thrombin-induced platelet aggregation. SEM revealed that <i>X. strumarium</i> pretreatment markedly preserved the resting platelet morphology and inhibited collagen-induced activation and shape changes. Further, the granule secretion, integrin-αIIbβ3 signaling, and the MAPK and PI3K/Akt pathways were also concentration-dependently inhibited. The in vivo blood flow rate and mice survival were improved, and H&E staining further revealed a concentration-dependent prevention of arterial occlusion following <i>X. strumarium</i> treatment. <b>Conclusions:</b> Collectively, <i>X. strumarium</i> demonstrated potent antiplatelet and antithrombotic effects, improving blood flow and survival while preventing arterial occlusion.