Skip to main content
Log in

Inhibitory effects of oligopeptides from hen egg white on both human platelet aggregation and blood coagulation

  • Research Articles
  • Drug Actions
  • Published:
Archives of Pharmacal Research Aims and scope Submit manuscript

Abstract

Egg white proteins have many biological functions and substantial nutritional benefits when used as a food source; however, they also contain allergens such as ovalbumin, ovomucoid, and ovotransferrin. We prepared oligopeptides without allergens from hen egg whites via the use of several proteases, and assessed their effects on platelet aggregation and blood coagulation, known to both of which are known to be major risk factors in thrombogenesis. Egg white oligopeptides (EWOP) inhibited collagen-induced human platelet aggregation in a dose-dependent manner. Additionally, we attempted to determine whether cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), aggregation-inhibiting intracellular molecules, regulate EWOP-inhibited platelet aggregation. EWOP caused an increase in cAMP levels, but did not affect cGMP levels, which suggests that the anti-platelet activity of EWOP operates in a cAMP-dependent manner, rather than via a cGMP-dependent process, in collagen-induced platelet aggregation. In addition, EWOP induced a significantly prolonged prothrombin time (PT) as compared with the controls. These data show that EWOP inhibits the conversion of fibrinogen to fibrin in a plasmatic atmosphere on an extrinsic pathway. Accordingly, these findings suggest that EWOP may be an excellent candidate as a crucial inhibitor of platelet activation, and its anti-platelet effects appear to involve the inhibition of both platelet aggregation and blood coagulation within the cardiovascular system.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Beavo, J. A., Cyclic nucleotide phosphodiesterases: functional implications of multiple isoforms. Physiol. Rev., 75, 725–748 (1995).

    PubMed  CAS  Google Scholar 

  • Berger, P. B., Clopidogrel after coronary stenting. Curr. Intervent. Cardiol. Rep., 1, 263–269 (1999).

    Google Scholar 

  • Berridge, M. J., The interaction of cyclic nucleotides and calcium in the control of cellular activity. Adv. Cyclic Nucleotide. Res., 6, 1–98 (1975).

    PubMed  CAS  Google Scholar 

  • Cannon, C., Mccabe, C., Borzak, S., Henry, T. D., Tischler, M. D., Mueller, H. S., Feldman, R., Palmeri, S. T., Ault, K., Hamilton, S. A., Rothman, J. M., Novotny, W. F., and Braunwald, E., Randomized trial of an oral platelet glycoprotein IIb/IIIa antagonist, sibrafiban, in patients after an acute coronary syndrome: results of the TIMI12 trial. Circulation, 97, 340–349 (1998).

    PubMed  CAS  Google Scholar 

  • Cavallini, L., Coassin, M., Borean, A., and Alexandre, A., Prostacyclin and sodium nitroprusside inhibit the activity of the platelet inositol 1,4,5-trisphosphate receptor and promote its phosphorylation. J. Biol. Chem., 271, 5545–5551 (1996).

    Article  PubMed  CAS  Google Scholar 

  • Chang, T. S., Kim, H. M., Lee, K. S., Khil, L. Y., Mar, W. C., Ryu, C. K., and Moon, C. K., Thromboxane A2 synthase inhibition and thromboxane A2 receptor blockade by 2-[(4-cyanophenyl)amino]-3-chloro-1,4-naphthalenediaone (NQ-Y15) in rat platelets. Biochem. Pharmacol., 54, 259–268 (1997).

    Article  PubMed  CAS  Google Scholar 

  • Cho, H. J., Ham, H. S., Lee, D. S., and Park, H. J., Effects of proteins from hen egg yolk on human platelet aggregation and blood coagulation. Biol. Pharm. Bull., 26, 1388–1392 (2003).

    Article  PubMed  CAS  Google Scholar 

  • Cho, H. J., Ham, H. S., Lee, T. K., Jung, Y. J., Choi, S. A., Kang, H. C., and Park, H. J., Inhibitory effect of cordycepin on human platelet aggregation. J. Biomed. Lab. Sci., 10, 1–8 (2004).

    Google Scholar 

  • Clapp, L. H., Finney, P., Turcato, S., Tran, S., Rubin, L. J., and Tinker, A., Differential effects of stable prostacyclin analogs on smooth muscle proliferation and cyclic AMP generation in human pulmonary artery. Am. J. Respir. Cell Mol. Biol., 26, 194–201 (2002).

    PubMed  CAS  Google Scholar 

  • Cutton, P., Folts, J. D., and Freedman, J. E., Pharmacological control of platelet function. Pharmacol. Reviews, 44, 255–264 (2001).

    Article  CAS  Google Scholar 

  • Doni, M. G., Alexandre, A., Padoin, E., Francesconi, M. A., and Deana, R., Staurosporine-independent platelet aggregation induced by the calcium ionophore ionomycin is inhibited by prostacyclin and sodium nitroprusside and stimulated by adrenaline. Arch. Biochem. Biophys., 301, 431–438 (1993).

    Article  PubMed  CAS  Google Scholar 

  • Geiger, J., Nolte, C., Butt, E., Sage, S. O., and Walter, U., Role of cGMP and cGMP-dependent protein kinase in nitrovasodilator inhibition of agonist-evoked calcium elevation in human platelets. Proc. Natl. Acad. Sci., 89, 1031–1035 (1992).

    Article  PubMed  CAS  Google Scholar 

  • Golino, P., Buja, L. M., Ashton, J. H., Kulkarni, P., Taylor, A., and Willerson, J. T., Effect of thromboxane and serotonin receptor antagonists on intracoronary platelet deposition in dogs with experimentally stenosed coronary arteries. Circulation, 78, 701–711 (1998).

    Google Scholar 

  • Gutierrez, M. A., Takahashi, H., and Juneja, L. R., Nutritive evaluation of hen eggs. In Yamamoto, T., Juneja, L. R., Hatta, H., Kim, M. (Eds.), Their basic and applied science. CRC Press Inc, NW, USA, pp. 19–31, (1997).

    Google Scholar 

  • Heemskerk, J. W., Feijge, M. A., Sage, S. O., and Walter, U., Indirect regulation of Ca2+ entry by cAMP-dependent protein kinases and phospholipase C in rat platelets. Eur. J. Biochem., 223, 543–551 (1994).

    Article  PubMed  CAS  Google Scholar 

  • Hennekens, C. H., Buring, J. E., Sandercock, P., Collins, R., and Petto, R., Aspirin and other anti-platelet agents in the secondary and primary prevention of cardiovascular disease. Circulation, 80, 749–756 (1989).

    PubMed  CAS  Google Scholar 

  • Herbert, J. M., Frehel, K., and Valle, E., Clopidogrel: a novel anti-platelet and anti-thrombotic agent. Cardiovasc. Drug Rev., 11, 180–198 (1993).

    Article  CAS  Google Scholar 

  • Hoffman, D. R., Immunochemical identification of the allergens in egg white. J. Allergy Clin. Immunol., 71, 481 (1983).

    Article  PubMed  CAS  Google Scholar 

  • Johnson, P. C., Ware, J. A., and Salzman, E. W., Concurrent measurement of platelet ionized calcium concentration and aggregation: studies with the lumiaggregometer. Thromb. Res., 40, 435–443 (1985).

    Article  PubMed  CAS  Google Scholar 

  • Kaser-Glanzmann, R., Gerber, E., and Luscher, E. F., Regulation of the intracellular calcium level in human blood platelets: cyclic adenosine 3′, 5′-monophosphate dependent phosphorylation of a 22,000 dalton component in isolated Ca2+-accumulated vesicles. Biochim. Biophys. Acta. 558, 344–347 (1979).

    Article  PubMed  CAS  Google Scholar 

  • Kereiakes, D., Kleiman, N., Ferguson, J., Masud, A. R., Broderick, T. M., Abbottsmith, C. W., Runyon, J. P., Anderson, L. C., Anders, R. J., Dreiling, R. J., Hantsbarger, G. L., Bryzinski, B., and Topol, E. J., Pharmacodynamic efficacy, clinical safety, and outcomes after prolonged platelet glycoprotein IIb/IIIa receptor blockade with oral xemilofiban: results of a multicenter, placebo-controlled, randomized trial. Circulation, 98, 1268–1278 (1998).

    PubMed  CAS  Google Scholar 

  • King, S. B., Optimizing anti-platelet therapy in coronary interventions. Clin. Cardiol., 23(suppl 6), 8–13 (2000).

    Article  Google Scholar 

  • Laemmli, U. K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685 (1970).

    Article  PubMed  CAS  Google Scholar 

  • Langeland, T., A clinical and immunological study of allergy to hen’s egg white. II. Antigens in hen’s egg white studies by crossed immunoelectrophoresis. Allergy, 37, 323 (1982).

    Article  PubMed  CAS  Google Scholar 

  • Leoncini, G. and Signorello, M. G., N-ethylmaleimide inhibition of thrombin-induced platelet aggregation. Biochem. Pharmacol., 58, 1293–1299 (1999).

    Article  PubMed  CAS  Google Scholar 

  • Llorens, S., Jordan, J., and Nava, E., The nitric oxide pathway in the cardiovascular system. J. Physiol. Biochem., 58, 179–188 (2002).

    Article  PubMed  CAS  Google Scholar 

  • Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J., Protein measurement with the Folin phenol reagent. J. Biol. Chem., 193, 265–275 (1951).

    PubMed  CAS  Google Scholar 

  • Marin, J. Q., Dermot, C. J., and Brendan, F., Glycoprotein IIb/IIIa receptor number and occupancy during chronic administration of an oral antagonist. J. Pharmcol. Exp. Ther., 295, 670–676 (2000).

    Google Scholar 

  • Maurice, D. H. and Haslam, R. J., Molecular basis of the synergistic inhibition of platelet function by nitrovasodilators and activators of adenylate cyclase: inhibition of cyclic AMP breakdown by cyclic GMP. Mol. Pharmacol., 37, 671–681 (1990).

    PubMed  CAS  Google Scholar 

  • Mullershausen, F., Friebe, A., Feil, R., Thompson, W. J., Hofmann, F., and Koesling, D., Direct activation of PDE5 by cGMP: long-term effects within NO/cGMP signaling. J. Cell Biol., 160, 719–727 (2003).

    Article  PubMed  CAS  Google Scholar 

  • Mustard, J. F. and Packham, M. A., Factors influencing platelet function: adhesion, release, and aggregation. Pharmacol. Reviews, 22, 97–187 (1970).

    CAS  Google Scholar 

  • Naka, Y., Roy, D. K., Liao, H., Chowdhury, N. C., Michler, R. E., Oz, M. C., and Pinsky, D. J., cAMP-mediated vascular protection in an orthotopic rat lung transplant model. Insights into the mechanism of action of prostaglandin E1 to improve lung preservation. Circ. Res., 79, 773–783 (1996).

    PubMed  CAS  Google Scholar 

  • Nakamura, K., Kimura, M., and Aviv, A., Role of cyclic nucleotides in store-mediated external Ca2+ entry in human platelets. Biochem. J., 310, 263–269 (1995).

    PubMed  CAS  Google Scholar 

  • Oh-ishi, S., Utsunomiya, I., Yamamoto, T., Komuro, Y., and Hara, Y., Effects of prostaglandins and cyclic AMP on cytokine production in rat leukocytes. Eur. J. Pharmacol., 300, 255–259 (1996).

    Article  PubMed  CAS  Google Scholar 

  • Paolucci, C., Burastero, S. E., Rovere-querinin, P., Palma, C. D., Falcone, S., Perrotta, C., Capoblanco, A., Manfredi, A. A., and Clement, E., Synergism of nitric oxide and maturation signals on human dendritic cells occurs through a cyclic GMP-dependent pathway. J. Leukoc. Biol., 73, 253–262 (2003).

    Article  PubMed  CAS  Google Scholar 

  • Rittenhouse-Simmons, S. and Deykin, D., Isolation of membranes from normal and thrombin-treated gel-filtered platelets using a lectin marker. Biochim. Biophys. Acta., 426, 688–696 (1976).

    Article  PubMed  CAS  Google Scholar 

  • Schwartz, S. M., heinmark, R. L., and Majesky, M. W., developmental mechanisms underlying pathology of arteries. Physiol. Rev., 72, 1177–1209 (1990).

    Google Scholar 

  • Schwarz, U. R., Walter, U., and Eigenthaler, M., Taming platelets with cyclic nucleotides. Biochem. Pharmacol., 62, 1153–1161 (2001).

    Article  PubMed  CAS  Google Scholar 

  • Sinha, A. K., Rao, A. K., Willis, J., and Colman, R. W., Inhibition of thromboxane A2 synthesis in human platelets by coagulation factor Xa. Proc. Natl. Acad. Sci., 80, 6086–6090 (1983).

    Article  PubMed  CAS  Google Scholar 

  • Smith, J. B. and Willis, A. L., Aspirin selectively inhibits prostaglandin production in human platelets. Nat. New. Biol., 231, 235–237 (1971).

    PubMed  CAS  Google Scholar 

  • Stones, J. R. and Marletta, M. A., Spectral and kinetic studies on the activation of soluble guanylate cylase by nitric oxide. Biochemistry, 35, 1093–1099 (1996).

    Article  Google Scholar 

  • Tertyshnikova, S. and Fein, A., Inhibition of inositol 1,4,5-trisphosphate-induced Ca2+ release by cAMP-dependent protein kinase in a living cell. Proc. Natl. Acad. Sci., 95, 1613–1617 (1998).

    Article  PubMed  CAS  Google Scholar 

  • Yamagami, S., Miyauchi, K., Kimura, T., Goh, Y., Daida, H., and Yamaguchi, H., Effects of the thromboxane A2 receptor antagonist on platelet deposition and intimal hyperplasia after balloon injury. Jpn. Heart. J., 40, 791–802 (1999).

    Article  PubMed  CAS  Google Scholar 

  • Yao, S. K., Ober, J. C., and McNatt, J., ADP plays an important role in mediating platelet aggregation and cyclic flow variations in vivo in stenosed and endothelium-injured canine coronary arteries. Circ. Res., 70, 39–48 (1992).

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Hwa-Jin Park.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cho, HJ., Kittaka, R., Abdou, A.M. et al. Inhibitory effects of oligopeptides from hen egg white on both human platelet aggregation and blood coagulation. Arch. Pharm. Res. 32, 945–953 (2009). https://doi.org/10.1007/s12272-009-1618-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12272-009-1618-y

Key words

Navigation