Abstract
OBJECTIVES: This study investigated the antithrombotic properties of a fibrinolytic enzyme (CFE) purified from the culture supernatant of Coprinus comatus using a zebrafish thrombosis model. METHODS: A phenylhydrazine-induced thrombosis model was employed to evaluate the in vivo thrombolytic efficacy and mechanisms of CFE. RESULTS: CFE significantly attenuated thrombogenesis by inhibiting erythrocyte aggregation in the caudal vessels, reducing staining intensity (3.61-fold decrease) and staining area (3.89-fold decrease). Concurrently, CFE enhanced cardiac hemodynamics, increasing erythrocyte staining intensity (9.29-fold) and staining area (5.55-fold) while achieving an 85.19% thrombosis inhibition rate. Behavioral analysis confirmed improved motility, with CFE-treated zebrafish exhibiting 2.23-fold increases in total movement distance and average speed, alongside a 3.59-fold extension in active movement duration. Mechanistically, ELISA revealed the multi-pathway activity of CFE, promoting fibrinolysis through reductions in plasminogen, fibrinogen, and D-dimer; inhibiting platelet activation via downregulation of prostaglandin-endoperoxide synthase (PTGS), thromboxane A2 (TXA2), P-selectin, and von Willebrand factor (vWF); and modulating coagulation cascades through elevated protein C and tissue factor pathway inhibitor (TFPI) with concurrent suppression of coagulation factor VII (FVII). CONCLUSIONS: These results indicate that the fibrinolytic enzyme CFE, derived from Coprinus comatus, exerts potent antithrombotic effects, supporting its potential as a basis for fungal-derived natural antithrombotic functional food ingredients.