Ile-Pro-Pro

Aims: Milk casein-derived bioactive tripeptides isoleucine-proline-proline (Ile-Pro-Pro) and valine-proline-proline (Val-Pro-Pro) lower blood pressure in animal models of hypertension and humans. In some studies, their angiotensin-converting enzyme (ACE)-inhibitory effect has been demonstrated. Besides classical ACE-angiotensin II-AT(1)-receptor pathway (ACE-Ang II- AT(1)), the significance of ACE2-angiotensin-(1-7)-Mas-receptor (ACE2-Ang-(1-7)-Mas) axis in the blood pressure regulation has now been acknowledged. The present study was aimed to further evaluate the renin-angiotensin system (RAS)-related vascular effects of Ile-Pro-Pro in vitro using rat mesenteric arteries. Main methods: Superior mesenteric arteries of spontaneously hypertensive rat (SHR) were isolated, cut into rings and mounted in standard organ bath chambers. Endothelium-intact arterial rings were incubated in Krebs solution either with Ile-Pro-Pro, proline-proline (Pro-Pro), isoleucine (Ile), proline (Pro) or captopril for 6h at +37°C and vascular reactivity was measured. Key findings: In the presence of AT(1)-antagonist valsartan, Ang II induced vasodilatation, which was more pronounced in the arteries incubated with Ile-Pro-Pro (P<0.05) compared to the other compounds. Ang-(1-7)-induced vasodilatation was augmented by Ile-Pro-Pro or Pro (P<0.001 vs. control). Mas-receptor antagonist A-779 did not alter the responses. Ile-Pro-Pro and Pro augmented also bradykinin-induced relaxations (P<0.001 vs. control). Control arteries and arteries incubated with captopril showed only slight relaxations at higher bradykinin concentrations. Significance: Casein-derived tripeptide Ile-Pro-Pro and amino acid Pro enhance the vasodilatory effect of Ang-(1-7) and bradykinin. The role of ACE2-Ang-(1-7)-Mas axis in the modulation of vascular tone by these compounds seems probable.

Much clinical evidence on the antihypertensive effects of the milk-derived antihypertensive peptides Val-Pro-Pro and Ile-Pro-Pro (lactotripeptides) has been reported. However, circadian rhythm effects determined by ambulatory blood pressure monitoring (ABPM) to eliminate the confounding influence of the white-coat effect have not been fully studied. Twelve hypertensive patients not receiving antihypertensive medication (2 men, 10 women; mean age±standard deviation, 63.5±8.3 years) who had been visiting our clinic for more than 1 year participated in this study. Mean (±standard deviation) systolic blood pressure (SBP) and diastolic blood pressure (DBP) were 142.4±2.6 and 83.5±6.4 mm Hg, respectively, at the first office visit. After patients ingested a fermented milk product containing antihypertensive peptides (2.53 mg Val-Pro-Pro; 1.52 mg Ile-Pro-Pro) for more than 4 weeks, both office SBP and DBP were significantly reduced to a mean (±standard deviation) of 133.3±7.0 mm Hg and 76.5±8.4 mm Hg (P<.001 and P<.005 by paired t-test), respectively. The 24-hour SBP and DBP determined by ABPM were reduced from 127.3±2.4 and 78.7±2.3 mm Hg to 120.2±2.4 and 75.0±2.2 mm Hg (P<.001 and P<.05), respectively. Awake-time SBP (08:00-21:00), night-time SBP (0:00-05:00), and early-morning SBP (06:00-07:00) were reduced from 130.9±2.4 to 123.3±2.3 mm Hg, 118.7±2.9 to 113.2±3.4 mm Hg, and 132.8±4.3 to 122.4±3.9 mm Hg (by paired t-test: P<.001, P<.05, and P<.05), respectively. As seen with DBP measured by ABPM, 24-hour DBP and awake-time DBP were significantly reduced from 78.7±2.3 to 75.0±2.2 mm Hg and 82.1±2.5 to 77.3±2.2 mm Hg (P<.05 and P<.01), respectively. Office BP and 24-hour blood pressure did not significantly differ between the dipper and nondipper groups at baseline. However, after treatment, night-time and early-morning blood pressure were significantly reduced from baseline in the nondipper group (-8.5±2.5 and -15.6±3.7 mm Hg; P<.05 and P<.01, respectively) but not in the dipper group (-2.5±3.6 and -1.2±4.7 mm Hg; P not significant), and the reduction in early-morning blood pressure significantly differed between the groups (P<.05). These results suggest that Val-Pro-Pro and Ile-Pro-Pro decrease blood pressure in patients with stage I hypertension and result not only in lower blood pressure at night-time but also in lower early-morning SBP in nondipper patients.

Angiotensin II (Ang II), a vasoactive factor in the renin-angiotensin-aldosterone system (RAAS), can regulate vasoconstriction and promote multiple vascular diseases. In this study, the effects of potent antihypertensive peptide Val-Pro-Pro (VPP) and Ile-Pro-Pro (IPP) on the proliferation and migration of vascular smooth muscle cells (VSMCs) by extracellular vesicles (EVs) from vascular endothelial cells (VECs) were studied using a cell co-culture model. The VEC-derived EVs were isolated, characterized, and investigated. The present study demonstrated that the EVs from Ang II-induced VECs could promote proliferation, migration, and inflammatory factors (IL-6 increased to 40.75 ± 4.33 pg/mL and IL-1β increased to 28.62 ± 5.42 pg/mL) generation of VSMCs, VPP and IPP exerted discrepant inhibitory effects on this pathway. The EVs with RNase treatment lost the effects on VSMCs, indicating that the RNAs packed into vesicles may be a critical component. These results implied that VPP and IPP could alleviate Ang II-induced vascular dysfunction by modulating the EV-mediated transmission of RNAs between VECs and VSMCs.