Tripeptide Hyp-Asp-Gly from collagen peptides inhibited platelet activation via regulation of PI3K/Akt-MAPK/ERK1/2 signaling pathway.

Platelet activation is involved in cardiovascular thrombosis. Our previous study demonstrated that oral administration of collagen peptides (CPs) inhibited platelet activation, but the mechanism of action of CPs remained to be elucidated. As a continued effort, the objective of this study was to identify the active ingredient of CPs and clarify its molecular mechanism. Simulated absorbate of CPs was prepared by simulated gastrointestinal digestion and intestinal absorption system, and then separated by C18 column. The fraction with the highest antiplatelet activity was subjected to NanoUPLC-ESI-MS/MS for peptide sequencing. Novel tripeptide Hyp-Asp-Gly (ODG) was identified. It had a broad-spectrum inhibition of platelet activation induced by collagen, thrombin, and adenosine diphosphate (ADP). ODG could survive simulated gastrointestinal digestion and be absorbed intact. Furthermore, it showed good stability in plasma. ODG had no significant effect on the PLC-PKC-Ca pathway, but it inhibited the PI3K/Akt-MAPK/ERK1/2 signaling. At a dosage of 200 µmol/kg body weight, ODG had an in vivo anti-thrombosis activity without bleeding risk. The present study provides one of the mechanisms of action of CPs and highlights its potential use as a functional component to combat cardiovascular thrombosis. PRACTICAL APPLICATION: This study has suggested that tripeptide Hyp-Asp-Gly(ODG) derived from collagen have potent activities. This novel collagen peptide had a greatpotential to be applied to combat cardiovascular thrombosis in the foodindustry. Meanwhile, this work is expected to provide a theoretical basis forthe development of safe and effective anti-platelet and anti-thrombosis peptides.