Proteomic approaches for the identification of novel cardiovascular biomarkers: A proof of concept

Introduction: Bioactive peptides have been defined as proteinderived short peptides that are released by hydrolysis processes such as dry-curing ham. Evidence suggests that bioactive peptides reduce hypertension and might affect other metabolic pathways which should be confirmed to validate its beneficial effects in humans. Objectives: To identify novel biomarkers of the cardiovascular status with promising proteomic techniques. Methods: Nine volunteers (49 ± 6 years old, 75% male) consumed dry-cured pork ham enriched in characterized antihypertensive peptides for 28 days. Protein quantification from all citrated plasma samples was performed using the Pierce BCA Protein Assay Kit followed by SDS-PAGE. The digested proteins were analysed with MS/MS using Sequest and X! Tandem Softwares. Peptide and protein identifications was validated though Scaffold Software. Then, immunohistochemical approaches (ELISA) were used to confirm these results. Results: The extended proteomic study identified changes in proteins such as Akt, MMP-8, MAPK, ApoB, ApoA2 and ApoA1 which are all related to cardiovascular mechanisms of action. Then, these results were confirmed by immunohistochemical techniques (p