Elucidating the Anti-thrombotic Mechanism of Equiseti hiemalis Herba through Integrated Pharmacology

This study systematically elucidates the anti-thrombotic material basis, potential targets, and molecular mechanisms of Equiseti hiemalis herba (EH) using an integrated strategy combining metabolomics, serum pharmacochemistry, network pharmacology, bioinformatics analyses, and molecular docking.

Components in the EH decoction were identified by UHPLC-MS. Sixteen normal male SD rats were administered EH to monitor weight and organ indices. Plasma metabolomics were used to analyze differential metabolites and pathways. Blood-absorbed prototype components were identified, and their targets were predicted. Network pharmacology was employed to intersect drug targets with thrombosis-related targets from GeneCards and an independent GEO dataset to identify core therapeutic targets. GO analysis and molecular docking were further used for validation.

A total of 138 compounds, including caffeic acid, ferulic acid, and 13-docosenamide, were identified in EH. Administration to rats revealed only slight effects on the liver. EH primarily exerts its effects by modulating fatty acid, amino acid, and nucleotide metabolism. It acts on nine key targets (CA9, MAOA, TYMS, etc.) via 15 active components (gingerol, xanthosine, adenosine, etc.), influencing four pathways (glycine and serine metabolism, ammonia recycling, arginine and proline metabolism, pyrimidine metabolism) to achieve its anti-thrombotic effects. Importantly, the dysregulation of these nine key targets was significantly validated in an independent clinical venous thrombosis cohort from the GEO database.

This study proposes a multi-component, multi-target, and multi-pathway mechanism for EH against thrombosis and provides strong external clinical evidence supporting the disease relevance of the predicted targets. The findings offer a solid theoretical basis and valuable candidate resources for the future development of EH as an anti-thrombotic drug.