Anti‐Thrombotic Effect of Hemp Seed Peptides in Zebrafish by Dual Regulation of vegfr1 Signaling and Caspase‐3‐Mediated Apoptosis

This study aimed to investigate the antithrombotic activity of hemp seed peptides derived from trypsin hydrolysates and elucidate their molecular mechanisms using a ponatinib-induced zebrafish (Danio rerio) thrombosis model. Through ultrafiltration and LC-MS/MS analysis, we identified 1741 peptide sequences (< 3 kDa), with 67% distributed in the 0.5-1.5 kDa range. In vivo experiments demonstrated that 125 μg/mL hemp seed peptides significantly ameliorated ponatinib-induced thrombosis by increasing 16.55% cardiac erythrocyte staining intensity, enhancing 54.8% blood flow velocity, and restoring 11.2% intersegmental blood vessel diameters. In addition, the observed anticoagulant effect may be partially attributed to upregulated expression levels of vascular endothelial growth factor receptor 1 (vegfr1) and down-regulated expression levels of cysteinyl aspartate-specific proteinase-3 (caspase-3) at the mRNA level. These findings propose hemp seed peptides as novel dietary antithrombotic agents with potential applications in functional food development.