永利3044(中国集团)有限公司官网

Abstract

Ethnopharmacological relevance Typhae Pollen (TP) has long been used in traditional Chinese medicine to promote blood circulation and resolve blood stasis, particularly in the management of thrombotic disorders. However, its bioactive constituents and antithrombotic mechanisms remain incompletely understood. Aim of the study This study aimed to systematically characterize the chemical constituents of TP and to elucidate its antithrombotic activity and underlying mechanisms. Materials and methods Comprehensive phytochemical profiling of TP was performed using UHPLC–QTOF–MS/MS combined with diagnostic fragment ion (DFI) and neutral loss (NL) filtering strategies. Multivariate statistical analysis was applied to evaluate chemical consistency and regional variation among TP samples. Antithrombotic activity was assessed using thrombin inhibition assays, ADP-induced platelet aggregation assays, and zebrafish thrombosis models. Molecular docking was conducted to analyze interactions between active constituents and thrombin. The effects of TP-derived compounds on inflammatory cytokines, apoptosis-related proteins, and adhesion molecules were further investigated. Results UHPLC–QTOF–MS/MS analysis identified 70 compounds, including flavonoids, organic acids, and other compound classes in TP, with overall chemical consistency despite regional differences. TP extract exhibited significant thrombin inhibitory activity, with seven organic acids showing notable effects (IC 50 = 0.209-0.457 mg/mL), and molecular docking revealed favorable binding affinities with thrombin (−5.20 to −6.17 kcal/mol). TP extract, p -coumaric acid, succinic acid, and citric acid significantly inhibited ADP-induced platelet aggregation in vitro . In zebrafish models, specific organic acids (catechin and p -coumaric acid), and flavonoids (rutin and hyperoside) markedly reduced arachidonic acid–induced cardiac red blood cell accumulation. Network pharmacology analysis suggested that the antithromboti