高分辨率肽组学揭示蚯蚓源14-3-3ζ靶向抗血小板肽用于心血管功能性食品

Abstract

Edible earthworm ( Pheretima aspergillum ), a promising sustainable protein source, demonstrate significant antithrombotic bioactivity in vivo upon oral administration. However, most reported antithrombotic components from earthworms are thermolabile macromolecular plasminases, which completely lose biological activity after conventional heat treatment, severely limiting their application in heat-processed food systems. In this study, we identified and characterized antithrombotic peptides from heat-processed earthworm, and deciphered their action mechanism via a bioactivity-guided fractionation approach combined with LC-MS/MS-based peptidomics and target fishing technology. The results revealed that the <3 kDa ultrafiltration fraction showed predominant antiplatelet efficacy, establishing it as the key bioactive component. Further purification using preparative liquid chromatography yielded seven distinct peptides, among which AGDGLKRVGGADDALDK (P1) and QQSQP (P6) exhibited the highest inhibitory potency against platelet aggregation in vitro. Furthermore, the earthworm active peptide P1 exhibits significant in vivo antithrombotic effects in animals without detectable bleeding risk. Target fishing and molecular docking technology revealed that P1 selectively targets 14–3-3ζ with an equilibrium dissociation constant (KD) of 6.08 μM, thereby exerting antiplatelet activity. These food-derived peptides demonstrate dual functionality as natural antiplatelet agents and microcirculation modulators, positioning earthworm peptides as a novel cardiovascular-health ingredient for functional foods targeting thrombosis prevention.