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Arzneimittelforschung 2012; 62(10): 477-481
DOI: 10.1055/s-0032-1321846
Original Article
© Georg Thieme Verlag KG Stuttgart · New York

Effects of Milk Casein Derived Tripeptides on Endothelial Enzymes In Vitro; a Study with Synthetic Tripeptides

A. Siltari
1   Institute of Biomedicine, Pharmacology, University of Helsinki, Helsinki, Finland
,
A. S. Kivimäki
1   Institute of Biomedicine, Pharmacology, University of Helsinki, Helsinki, Finland
,
P. I. Ehlers
1   Institute of Biomedicine, Pharmacology, University of Helsinki, Helsinki, Finland
,
R. Korpela
1   Institute of Biomedicine, Pharmacology, University of Helsinki, Helsinki, Finland
,
H. Vapaatalo
1   Institute of Biomedicine, Pharmacology, University of Helsinki, Helsinki, Finland
› Author Affiliations
Further Information

Publication History

received 06 March 2012

accepted 04 July 2012

Publication Date:
23 August 2012 (online)

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Abstract

In the fermentation of milk by certain lactic acid bacteria, casein is degraded into bioactive tripeptides shown to lower blood pressure in experimental animal models and in mildly hypertensive humans. This effect is suggested to result mainly in inhibition of angiotensin converting enzyme 1 (ACE-1).

Due to the complexity of renin-angiotensin system (RAS), several other enzymes than ACE-1 can participate in the production of vasoactive components. Therefore, in the present study we investigated effects of tripeptides isoleucine-proline-proline (IPP), valine-proline-proline (VPP) and leucine-proline-proline (LPP) on some endothelial enzymes that are important in RAS or otherwise have a role in the endothelial function. The enzymes investigated were renin, chymase, neutral endopeptidase (NEP), prolyl oligopeptidase (POP), cathepsin G, endothelin converting enzyme 1 (ECE-1), and cyclooxygenase 1 and 2 (COX -1 and COX-2).

The tripeptides inhibited prolyl oligopeptidase (POP) dose-dependently. IPP was the most potent inhibitor (IC50 486±95 µM). Contrary, cathepsin G was activated by IPP, VPP and LPP as well as the amino acids proline and isoleucine. The other investigated enzymes were not affected. Inhibition of POP and activation of cathepsin G do not explain the blood pressure lowering effects of the tripeptides. Thus the inhibition of ACE-1 remains the most plausible mechanism of the antihypertensive effects of the tripeptides.

Key words

blood pressure - renin-angiotensin-system - endothelium - tripeptide - cathepsin G - prolyl oligopeptidase
 

  • References

  • 1 Varagic J, Trask AJ, Jessup JA et al. New angiotensins. J Mol Med 2008; 86: 663-671
    CrossrefPubMedGoogle Scholar
  • 2 Fyhrqvist F, Saijonmaa O. Renin-angiotensin system revisited. J Intern Med 2008; 264: 224-236
    CrossrefPubMedGoogle Scholar
  • 3 Katovich MJ, Grobe JL, Raizada MK. Angiotensin-(1-7) as an antihypertensive, antifibrotic target. Curr Hypertens Rep 2008; 10: 227-232
    CrossrefPubMedGoogle Scholar
  • 4 Cat AND, Touyz RMA. New look at the renin-angiotensin system- focusing on the vasculature system. Peptides 2011; 32: 2141-2150
    CrossrefPubMedGoogle Scholar
  • 5 Ehlers PI, Nurmi L, Turpeinen AM et al. Casein-derived tripeptide Ile-Pro-Pro improves angiotensin-(1-7)- and bradykinin-induced rat mesenteric artery relaxation. Life Sci 2011; 88: 206-211
    CrossrefPubMedGoogle Scholar
  • 6 Nakamura Y, Yamamoto N, Sakai K et al. Purification and characterization of angiotensin I-converting enzyme inhibitors from sour milk. J Dairy Sci 1995; 78: 777-783
    CrossrefPubMedGoogle Scholar
  • 7 Sipola M, Finckenberg P, Santisteban J et al. Long-term intake of milk peptide attenuates development of hypertension in spontaneously hypertensive rats. J Physiol Pharmacol 2001; 52: 745-754
    PubMedGoogle Scholar
  • 8 Jäkälä P, Jauhiainen T, Korpela R et al. Milk protein-derived bioactive tripeptides Ile-Pro-Pro and Val-Pro-Pro protect endothelial function in vitro in hypertensive rats. J Functional Foods 2009; 1: 266-273
    CrossrefPubMedGoogle Scholar
  • 9 Jäkälä P, Pere E, Lehtinen R et al. Cardiovascular activity of milk casein-derived tripeptides and plant sterols in spontaneously hypertensive rats. J Physiol Pharmacol 2009; 60: 11-20
    PubMedGoogle Scholar
  • 10 Pripp AH. Effect of peptides derived from food proteins on blood pressure: a meta-analysis of randomized controlled trials. Food Nutr Res 2008; 32 DOI: 10.3402/fnr.v52i0.1641.
    PubMedGoogle Scholar
  • 11 Xu JY, Qin LQ, Wang PY et al. Effects of milk tripeptides on blood pressure: a meta-analysis of randomized controlled trials. Nutrition 2008; 24: 933-940
    CrossrefPubMedGoogle Scholar
  • 12 Cicero AF, Gerocarni B, Laghi L et al. Blood pressure lowering effects of lactotripeptides assumed as functional food: a meta-analysis of current available clinical trials. J Hum Hypertens 2011; 25: 425-436
    CrossrefPubMedGoogle Scholar
  • 13 Lehtinen R, Jauhiainen T, Kankuri E et al. Effects of milk casein-derived tripeptides Ile-Pro-Pro, Val-Pro-Pro, and Leu-Pro-Pro on enzymes processing vasoactive precursors in vitro. Aznelmittelforschung 2010; 60: 182-185
    PubMedGoogle Scholar
  • 14 Florentin D, Sassi A, Roques BP. A highly sensitive fluorometric assay for enkephalinase, a neutral metallopeptidase that releases tyrosine-glycine-glycine from enkephalins. Anal Biochem 1984; 141: 62-69
    CrossrefPubMedGoogle Scholar
  • 15 Atack JR, Suman-Chauhan N, Dawson G et al. In vitro and in vivo inhibition of prolyl endopeptidase. Eur J Pharm 1991; 205: 157-163
    CrossrefPubMedGoogle Scholar
  • 16 Stanziola L, Greene LJ, Santos RA. Effect of chronic angiotensin converting enzyme inhibition on angiotensin I and bradykinin metabolism in rats. Am J Hypert 1999; 12: 1021-1029
    PubMedGoogle Scholar
  • 17 Barrett AJ, Cathepsin G. Meth in Enzym 1981; 80: 561-565
    PubMed
  • 18 Greene LJ, Spadaro AC, Martins A et al. Brain endo-oligopeptidase B: a post-proline cleaving enzyme that inactivates angiotensin I and II. Hypertension 1982; 4: 178-184
    CrossrefPubMedGoogle Scholar
  • 19 Ward PE, Bausback HH, Odya CE. Kinin and angiotensin metabolism by purified renal post-proline cleaving enzymes. Biochem Pharmacol 1987; 36: 3187-3193
    CrossrefPubMedGoogle Scholar
  • 20 Jauhiainen T, Wuolle K, Vapaatalo H et al. Oral absoption, tissue distribution and excretion of a radiolabelled analog of a milk-derived antihypertensive peptide, Ile-Pro-Pro, in rats. Int Dairy J 2007; 17: 1216-1223
    CrossrefPubMedGoogle Scholar
  • 21 Koida M, Walter R. Post-proline cleaving enzyme. Purification of this endopeptidase by affinity chromatography. J Biol Chem 1976; 251: 7593-7599
    PubMedGoogle Scholar
  • 22 Yoshimoto T, Nishimura T, Takatsuka N et al. Post-proline cleaving enzyme (prolyl endopeptidase) from bovine brain. J Biochem 1983; 94: 1179-1190
    PubMedGoogle Scholar
  • 23 Stanziola L, Greene LJ, Santos RAS. Effect of chronic angiotensin converting enzyme inhibition on angiotensin I and bradykinin metabolism in rats. Am J Hypertens 1999; 12: 1021-1029
    CrossrefPubMedGoogle Scholar
  • 24 Lawandi J, Gerber-Lemaire S, Juillerat-Jeanneret L et al. Inhibitors of prolyl oligopeptidases for the therapy of human disease: Defining diseases and inhibitors. J Med Chem 2010; 53: 3423-3438
    CrossrefPubMedGoogle Scholar
  • 25 Carcia-Horsman JA, Männistö PT, Venäläinen JI. On the role of prolyl oligopeptidase on health and disease. Neuropeptides 2007; 41: 1-24
    CrossrefPubMedGoogle Scholar
  • 26 Brecher AS, Dubord R. Effect on acetaldehyde upon cathepsin G and chymase. NRAS implications. Dig Dis Sci 2008; 53: 1311-1315
    CrossrefPubMedGoogle Scholar