Skip to main content

Antithrombotic activities of aspalathin and nothofagin via inhibiting platelet aggregation and FIIa/FXa

Abstract

Aspalathin (Asp) and nothofagin (Not) are two major active dihydrochalcones found in green rooibos tea (Aspalathus linearis; family, Fabaceae; tribe, Crotalarieae), which have been reported for their anti-oxidant activity. Here, the anticoagulant activities of Asp and Not were examined by monitoring activated partial thromboplastin time (aPTT), prothrombin time (PT), and the activities of thrombin (Factor IIa, FIIa) and activated factor X (FXa). And, the effects of Asp and Not on expression of plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) were evaluated in tumor necrosis factor (TNF)-α activated human umbilical vein endothelial cells (HUVECs). Treatment with Asp and Not resulted in prolonged aPTT and PT and inhibition of the activities of thrombin and FXa, as well as inhibited production of thrombin and FXa in HUVECs. In addition, Asp and Not inhibited thrombin-catalyzed fibrin polymerization and platelet aggregation. Asp and Not also elicited anticoagulant effects in mice. In addition, treatment with Asp and Not resulted in significant reduction of the PAI-1 to t-PA ratio. Collectively, Asp and Not possesses antithrombotic activities and offers a basis for development of a novel anticoagulant.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3

References

  • Altieri, D.C., P.M. Mannucci, and A.M. Capitanio. 1986. Binding of fibrinogen to human monocytes. Journal of Clinical Investigation 78: 968–976.

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  • Bae, J.S. 2011. Antithrombotic and profibrinolytic activities of phloroglucinol. Food and Chemical Toxicology 49: 1572–1577.

    CAS  PubMed  Article  Google Scholar 

  • Bae, J.S., and A.R. Rezaie. 2008. Protease activated receptor 1 (PAR-1) activation by thrombin is protective in human pulmonary artery endothelial cells if endothelial protein C receptor is occupied by its natural ligand. Thrombosis and Haemostasis 100: 101–109.

    CAS  PubMed Central  PubMed  Google Scholar 

  • Dahlback, B. 2000. Blood coagulation. Lancet 355: 1627–1632.

    CAS  PubMed  Article  Google Scholar 

  • Davie, E.W. 1995. Biochemical and molecular aspects of the coagulation cascade. Thrombosis and Haemostasis 74: 1–6.

    CAS  PubMed  Google Scholar 

  • Davie, E.W., K. Fujikawa, and W. Kisiel. 1991. The coagulation cascade: initiation, maintenance, and regulation. Biochemistry 30: 10363–10370.

    CAS  PubMed  Article  Google Scholar 

  • Dejana, E., A. Callioni, A. Quintana, and G. De Gaetano. 1979. Bleeding time in laboratory animals. II: A comparison of different assay conditions in rats. Thrombosis Research 15: 191–197.

    CAS  PubMed  Article  Google Scholar 

  • Despotis, G.J., G. Gravlee, K. Filos, and J. Levy. 1999. Anticoagulation monitoring during cardiac surgery: a review of current and emerging techniques. Anesthesiology 91: 1122–1151.

    CAS  PubMed  Article  Google Scholar 

  • Esmon, C.T. 2005. The interactions between inflammation and coagulation. British Journal of Haematology 131: 417–430.

    CAS  PubMed  Article  Google Scholar 

  • Fareed, J., I. Thethi, and D. Hoppensteadt. 2012. Old versus new oral anticoagulants: focus on pharmacology. Annual Review of Pharmacology and Toxicology 52: 79–99.

    CAS  PubMed  Article  Google Scholar 

  • Fares, A. 2013. Winter cardiovascular diseases phenomenon. North American Journal of Medical Sciences 5: 266–279.

    PubMed Central  PubMed  Article  Google Scholar 

  • Ghosh, S., M. Ezban, E. Persson, U. Pendurthi, U. Hedner, and L.V. Rao. 2007. Activity and regulation of factor VIIa analogs with increased potency at the endothelial cell surface. Journal of Thrombosis and Haemostasis 5: 336–346.

    CAS  PubMed  Article  Google Scholar 

  • Hamaguchi, E., T. Takamura, A. Shimizu, and Y. Nagai. 2003. Tumor necrosis factor-alpha and troglitazone regulate plasminogen activator inhibitor type 1 production through extracellular signal-regulated kinase- and nuclear factor-kappaB-dependent pathways in cultured human umbilical vein endothelial cells. Journal of Pharmacology and Experimental Therapeutics 307: 987–994.

    CAS  PubMed  Article  Google Scholar 

  • Harbrecht, U. 2011. Old and new anticoagulants. Hamostaseologie 31: 21–27.

    CAS  PubMed  Article  Google Scholar 

  • Jauch, E.C., J.L. Saver, H.P. Adams Jr, A. Bruno, J.J. Connors, B.M. Demaerschalk, P. Khatri, P.W. Mcmullan Jr, A.I. Qureshi, K. Rosenfield, P.A. Scott, D.R. Summers, D.Z. Wang, M. Wintermark, and H. Yonas. 2013. Guidelines for the early management of patients with acute ischemic stroke: a guideline for healthcare professionals from the American Heart Association/American Stroke Association. Stroke 44: 870–947.

    PubMed  Article  Google Scholar 

  • Joubert, E., W.C. Gelderblom, A. Louw, and D. De Beer. 2008. South African herbal teas: Aspalathus linearis, Cyclopia spp. and Athrixia phylicoides–a review. Journal of Ethnopharmacology 119: 376–412.

    CAS  PubMed  Article  Google Scholar 

  • Kazuno, S., M. Yanagida, N. Shindo, and K. Murayama. 2005. Mass spectrometric identification and quantification of glycosyl flavonoids, including dihydrochalcones with neutral loss scan mode. Analytical Biochemistry 347: 182–192.

    CAS  PubMed  Article  Google Scholar 

  • Kim, S.Y., S. Kim, J.M. Kim, E.H. Jho, S. Park, D. Oh, and H.S. Yun-Choi. 2011. PKC inhibitors RO 31-8220 and Go 6983 enhance epinephrine-induced platelet aggregation in catecholamine hypo-responsive platelets by enhancing Akt phosphorylation. BMB Reports 44: 140–145.

    CAS  PubMed  Article  Google Scholar 

  • Kim, T.H., S.K. Ku, and J.S. Bae. 2012. Antithrombotic and profibrinolytic activities of eckol and dieckol. Journal of Cellular Biochemistry 113: 2877–2883.

    CAS  PubMed  Article  Google Scholar 

  • Krafczyk, N., F. Woyand, and M.A. Glomb. 2009. Structure-antioxidant relationship of flavonoids from fermented rooibos. Molecular Nutrition & Food Research 53: 635–642.

    CAS  Article  Google Scholar 

  • Lee, K.H. 2000. Research and future trends in the pharmaceutical development of medicinal herbs from Chinese medicine. Public Health Nutrition 3: 515–522.

    CAS  PubMed  Article  Google Scholar 

  • Lopez, S., F. Peiretti, B. Bonardo, I. Juhan-Vague, and G. Nalbone. 2000. Effect of atorvastatin and fluvastatin on the expression of plasminogen activator inhibitor type-1 in cultured human endothelial cells. Atherosclerosis 152: 359–366.

    CAS  PubMed  Article  Google Scholar 

  • Mann, K.G., K. Brummel, and S. Butenas. 2003. What is all that thrombin for? Journal of Thrombosis and Haemostasis 1: 1504–1514.

    CAS  PubMed  Article  Google Scholar 

  • Mckay, D.L., and J.B. Blumberg. 2007. A review of the bioactivity of South African herbal teas: rooibos (Aspalathus linearis) and honeybush (Cyclopia intermedia). Phytotherapy Research 21: 1–16.

    CAS  PubMed  Article  Google Scholar 

  • Nakamura, Y., S. Watanabe, N. Miyake, H. Kohno, and T. Osawa. 2003. Dihydrochalcones: evaluation as novel radical scavenging antioxidants. Journal of Agriculture and Food Chemistry 51: 3309–3312.

    CAS  Article  Google Scholar 

  • Nowak, P., H.M. Zbikowska, M. Ponczek, J. Kolodziejczyk, and B. Wachowicz. 2007. Different vulnerability of fibrinogen subunits to oxidative/nitrative modifications induced by peroxynitrite: functional consequences. Thrombosis Research 121: 163–174.

    CAS  PubMed  Article  Google Scholar 

  • Philip-Joet, F., M.C. Alessi, C. Philip-Joet, M. Aillaud, J.R. Barriere, A. Arnaud, and I. Juhan-Vague. 1995. Fibrinolytic and inflammatory processes in pleural effusions. European Respiratory Journal 8: 1352–1356.

    CAS  PubMed  Article  Google Scholar 

  • Quinn, C., J. Hill, and H. Hassouna. 2000. A guide for diagnosis of patients with arterial and venous thrombosis. Clinical Laboratory Science 13: 229–238.

    CAS  PubMed  Google Scholar 

  • Rao, L.V., S.I. Rapaport, and M. Lorenzi. 1988. Enhancement by human umbilical vein endothelial cells of factor Xa-catalyzed activation of factor VII. Blood 71: 791–796.

    CAS  PubMed  Google Scholar 

  • Rezk, B.M., G.R. Haenen, W.J. Van Der Vijgh, and A. Bast. 2002. The antioxidant activity of phloretin: the disclosure of a new antioxidant pharmacophore in flavonoids. Biochemical and Biophysical Research Communications 295: 9–13.

    CAS  PubMed  Article  Google Scholar 

  • Schleef, R.R., M.P. Bevilacqua, M. Sawdey, M.A. Gimbrone Jr, and D.J. Loskutoff. 1988. Cytokine activation of vascular endothelium. Effects on tissue-type plasminogen activator and type 1 plasminogen activator inhibitor. Journal of Biological Chemistry 263: 5797–5803.

    CAS  PubMed  Google Scholar 

  • Schouten, M., W.J. Wiersinga, M. Levi, and T. Van Der Poll. 2008. Inflammation, endothelium, and coagulation in sepsis. Journal of Leukocyte Biology 83: 536–545.

    CAS  PubMed  Article  Google Scholar 

  • Snijman, P.W., E. Joubert, D. Ferreira, X.C. Li, Y. Ding, I.R. Green, and W.C. Gelderblom. 2009. Antioxidant activity of the dihydrochalcones Aspalathin and Nothofagin and their corresponding flavones in relation to other Rooibos (Aspalathus linearis) Flavonoids, Epigallocatechin Gallate, and Trolox. Journal of Agriculture and Food Chemistry 57: 6678–6684.

    CAS  Article  Google Scholar 

  • Sugo, T., C. Nakamikawa, S. Tanabe, and M. Matsuda. 1995. Activation of prothrombin by factor Xa bound to the membrane surface of human umbilical vein endothelial cells: its catalytic efficiency is similar to that of prothrombinase complex on platelets. Journal of Biochemistry 117: 244–250.

    CAS  PubMed  Article  Google Scholar 

  • Weitz, J.I., J. Hirsh, and M.M. Samama. 2004. New anticoagulant drugs: the Seventh ACCP Conference on Antithrombotic and Thrombolytic Therapy. Chest 126: 265S–286S.

    CAS  PubMed  Article  Google Scholar 

Download references

Acknowledgments

This study was supported by the National Research Foundation of Korea (NRF) funded by the Korean government (MSIP) (Grant Nos. 2013-067053).

Conflict of interest

The authors have no conflict of interest to declare.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jong-Sup Bae.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Ku, SK., Lee, W., Kang, M. et al. Antithrombotic activities of aspalathin and nothofagin via inhibiting platelet aggregation and FIIa/FXa. Arch. Pharm. Res. 38, 1080–1089 (2015). https://doi.org/10.1007/s12272-014-0501-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12272-014-0501-7

Keywords

  • Aspalathin
  • Nothofagin
  • Coagulation cascade
  • Fibrinolysis
  • Endothelium