An in vivo study of the antihypertensive effects of the umami peptide AHSVRFY from Parma ham and its intestinal digestion.

AHSVRFY, an umami peptide derived from Parma ham with reported in vitro angiotensin-converting enzyme (ACE) inhibitory activity (IC50 = 16.3 ± 0.16 μM), lacks comprehensive in vivo validation. To address this, we established an "digestion-activation-delivery" hypothesis and systematically validated it. In spontaneously hypertensive rats (SHRs), a single oral dose of AHSVRFY induced a significant reduction in systolic blood pressure (SBP) (13.9 ± 2.7 mmHg, p < 0.01). UPLC-MS/MS analysis revealed its degradation into six peptides in the intestine, and four of them were identified as novel ACE inhibitors. The most potent dipeptide, FY (IC50 = 45.11 ± 15.34 μM), was absorbed intact into the systemic circulation of SHRs. Crucially, the antihypertensive effect was mechanistically linked to a significant improvement in vascular endothelial function, as evidenced by increased plasma nitric oxide (NO) and decreased endothelin-1 (ET-1) levels. Furthermore, FY showed a protective effect on human umbilical vein endothelial cells (HUVECs) through the modulation of NO/ET-1. Additionally, its stable binding to the ACE active site was confirmed via molecular docking and dynamics simulations. This work shows that AHSVRFY's antihypertensive effect originates from GI-derived bioactive peptides, establishing a "digestion-activation-delivery" framework for the development of multifunctional food ingredients.