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Bovine lactoferrin (LF) hydrolysates and peptides identified thereof have shown antihypertensive effects in rat models, mainly but not exclusively by angiotensin-converting enzyme inhibition. In this study we aimed to assess the vasoactive effects and mechanisms of an ultrafiltered (< 3 kDa) pepsin LF hydrolysate (LFH) and a heptapeptide identified in a LF hydrolysate produced by yeast proteolysis (DPYKLRP) in peripheral resistance arteries from spontaneously hypertensive rats (SHRs).We used a myograph system for isometric tension recording in isolated small mesenteric arteries from SHRs. Direct vasoactive effects of LFH (30–100 μg/mL) and DPYKLRP (30–100 μM) were assessed in arteries precontracted with phenylephrine (PE, 10 μM) or KCl (120 mM), and in PE-precontracted arteries preincubated (10 min) with the NO synthase inhibitor L-NAME (0.1 mM) or the cyclooxygenase inhibitor indomethacin (10 μM). Indirect vasoactive effects of LFH (30–100 μg/mL) or DPYKLRP (30–100 μM) preincubation (10 min) on the relaxant responses to the NO donor sodium nitroprusside (SNP, 0.01–10 μM) or acetylcholine (Ach, 1–100 μM) were also studied in PE-precontracted arteries.Both LHF and DPYKLRP elicited direct relaxation of mesenteric arteries, by a mechanism involving NO release, counteracting modulation by prostanoids and K+ efflux. Moreover, LF-derived peptides also showed indirect vasoactive effects by enhancing endothelium-dependent relaxation to Ach and endothelium-independent relaxation to SNP.In conclusion, LF-derived peptides show ex vivodirect and indirect relaxing effects in small mesenteric arteries from SHRs. These vasoactive effects would reduce vascular peripheral resistance in vivo, and thus contribute to their antihypertensive effects.