An <i>in vivo</i> 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 <i>in vitro</i> angiotensin-converting enzyme (ACE) inhibitory activity (IC₅₀ = 16.3 ± 0.16 μM), lacks comprehensive <i>in vivo</i> 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, <i>p</i> < 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 <i>via</i> 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.