Abstract
Arterial thrombosis is an important complication of diabetes and cancer, being an important target for therapeutic intervention. Crataeva tapia bark lectin (CrataBL) has been previously shown to have hypoglycemiant effect and also to induce cancer cell apoptosis. It also showed inhibitory activity against Factor Xa (Kiapp=8.6 μm). In the present study, we evaluated the anti-thrombotic properties of CrataBL in arterial thrombosis model. CrataBL prolongs the activated partial thromboplastin time on human and mouse plasma, and it impairs the heparin-induced potentiation of antithrombin III and heparin-induced platelet activation in the presence of low-dose ADP. It is likely that the dense track of positive charge on CrataBL surface competes with the heparin ability to bind to antithrombin III and to stimulate platelets. In the photochemically induced thrombosis model in mice, in the groups treated with 1.25, 5.0, or 10 mg/kg CrataBL, prior to the thrombus induction, the time of total artery occlusion was prolonged by 33.38%, 65%, and 66.11%, respectively, relative to the time of the control group. In contrast to heparin, the bleeding time in CrataBL-treated mice was no longer than in the control. In conclusion, CrataBL was effective in blocking coagulation and arterial thrombus formation, without increasing bleeding time.
Acknowledgments
We thank Dr. Alexander Wlodawer for critical reading of this manuscript. We acknowledge the support of Fundação de Amparo à Pesquisa do Estado de São Paulo Grant 2009/53766-5, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).
Conflict of interest statement: The authors declare that no conflict of interest exists.
References
Araujo, R.M., Ferreira, Rda S., Napoleao, T.H., Carneiro-da-Cunha, M., Coelho, L.C., Correia, M.T.S., Oliva, M.L., and Paiva, P.M. (2012). Crataeva tapia bark lectin is an affinity adsorbent and insecticidal agent. Plant Sci. 183, 20–26.10.1016/j.plantsci.2011.10.018Search in Google Scholar PubMed
Born, G.V. and Cross, M.J. (1963). The aggregation of blood platelets. J. Physiol. 168, 178–195.10.1113/jphysiol.1963.sp007185Search in Google Scholar PubMed PubMed Central
Colliec-Jouault, S., Millet, J., Helley, D., Sinquin, C., and Fischer, A.M. (2003). Effect of low-molecular-weight fucoidan on experimental arterial thrombosis in the rabbit and rat. Thromb. Haemost. 1, 1114–1115.10.1046/j.1538-7836.2003.t01-1-00215.xSearch in Google Scholar PubMed
Colman, R.W. (2006). Are hemostasis and thrombosis two sides of the same coin? J. Exp. Med. 203, 493–495.10.1084/jem.20060217Search in Google Scholar PubMed PubMed Central
da Rocha, A.A., Araujo, T.F., da Fonseca, C.S., da Mota, D.L., de Medeiros, P.L., Paiva, P.M., Coelho, L.C., Correia, M.T.S., and Lima, V.L. (2013). Lectin from Crataeva tapia bark improves tissue damages and plasma hyperglycemia in alloxan-induced diabetic mice. Evid. Based Complement Alternat. Med. 2013, 869305.10.1155/2013/869305Search in Google Scholar PubMed PubMed Central
de Paula, C.A., Coulson-Thomas, V.J., Ferreira, J.G., Maza, P.K., Suzuki, E., Nakahata, A.M., Nader, H.B., Sampaio, M.U., and Oliva, M.L. (2012). Enterolobium contortisiliquum trypsin inhibitor (EcTI), a plant proteinase inhibitor, decreases in vitro cell adhesion and invasion by inhibition of Src protein-focal adhesion kinase (FAK) signaling pathways. J. Biol. Chem. 287, 170–182.10.1074/jbc.M111.263996Search in Google Scholar PubMed PubMed Central
Ferreira, R.D., Zhou, D., Ferreira, J.G., Silva, M.C., Silva-Lucca, R.A., Mentele, R., Paredes-Gamero, E.J., Bertolin, T.C., Correia, M.T.S., Paiva, P.M., et al. (2013). Crystal structure of bark protein (CrataBL) and its effect in human prostate cancer cell lines. PLoS One 8, e64426.10.1371/journal.pone.0064426Search in Google Scholar PubMed PubMed Central
Gao, W., Lin, T., Li, T., Yu, M., Hu, X., and Duan, D. (2013). Sodium alginate/heparin composites on PVC surfaces inhibit the thrombosis and platelet adhesion: applications in cardiac surgery. Int. J. Clin. Exp. Med. 6, 259–268.Search in Google Scholar
Gozzo, A.J., Nunes, V.A., Cruz-Silva, I., Carmona, A.K., Nader, H.B., Faljoni-Alario, A., Sampaio, M.U., and Araujo, M.S. (2006). Heparin modulation of human plasma kallikrein on different substrates and inhibitors. Biol. Chem. 387, 1129–1138.10.1515/BC.2006.139Search in Google Scholar PubMed
Hampl, V., Waters, C.L., and Archer, S.L. (1996). Determination of nitric oxide by the chemiluminescence reaction with ozone. In: Methods in Nitric Oxide Research. M. Feelisch, J.S. Stamler, eds. (New York, United States: John Wiley and Sons, Ltd), pp. 309–318.Search in Google Scholar
He, L., Vicente, C.P., Westrick, R.J., Eitzman, D.T., and Tollefsen, D.M. (2002). Heparin cofactor II inhibits arterial thrombosis after endothelial injury. J. Clin. Invest. 109, 213–219.10.1172/JCI0213432Search in Google Scholar
Hirsh, J., Bauer, K.A., Donati, M.B., Gould, M., Samama, M.M., Weitz, J.I., and American College of Chest P. (2008). Parenteral anticoagulants: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines (8th Edition). Chest 133, 141S–159S.10.1378/chest.08-0782Search in Google Scholar PubMed
Macedo, M.L., Diz Filho, E.B., Freire, M.G., Oliva, M.L., Sumikawa, J.T., Toyama, M.H., and Marangoni, S. (2011). A trypsin inhibitor from Sapindus saponaria L. seeds: purification, characterization, and activity towards pest insect digestive enzyme. Protein J. 30, 9–19.10.1007/s10930-010-9296-7Search in Google Scholar PubMed
Oliva, M.L., Andrade, S.A., Juliano, M.A., Sallai, R.C., Torquato, R.J., Sampaio, M.U., Pott, V.J., and Sampaio, C.A. (2003). Kinetic characterization of factor Xa binding using a quenched fluorescent substrate based on the reactive site of factor Xa inhibitor from Bauhinia ungulata seeds. Curr. Med. Chem. 10, 1085–1093.10.2174/0929867033457548Search in Google Scholar PubMed
Paredes-Gamero, E.J., Medeiros, V.P., Farias, E.H., Justo, G.Z., Trindade, E.S., Andrade-Lopes, A.L., Godinho, R.O., de Miranda, A., Ferreira, A.T., Tersariol, I.L., et al. (2010). Heparin induces rat aorta relaxation via integrin-dependent activation of muscarinic M3 receptors. Hypertension 56, 713–721.10.1161/HYPERTENSIONAHA.110.156877Search in Google Scholar PubMed
Prandoni, P., Lensing, A.W., Piccioli, A., Bernardi, E., Simioni, P., Girolami, B., Marchiori, A., Sabbion, P., Prins, M.H., Noventa, F., et al. (2002). Recurrent venous thromboembolism and bleeding complications during anticoagulant treatment in patients with cancer and venous thrombosis. Blood 100, 3484–3488.10.1182/blood-2002-01-0108Search in Google Scholar PubMed
Prevention of cardiovascular disease Guideline for assessment and management of cardiovascular risk: WHO Consultation, Geneve 2007. Geneva, World Health Organization, 2007 (document available at http://www.who.int/cardiovascular_diseases/guidelines/PocketGL.ENGLISH.AFR-D-E.rev1.pdf?ua=1, accessed 02 February, 2014.Search in Google Scholar
Rivera, J., Lozano, M.L., Navarro-Nunez, L., and Vicente, V. (2009). Platelet receptors and signaling in the dynamics of thrombus formation. Haematologica 94, 700–711.10.3324/haematol.2008.003178Search in Google Scholar PubMed PubMed Central
Silva, M.C.C., Santana, L.A., Mentele, R, Ferreira, R.S., de Miranda, A., Silva-Lucca, R.A., Sampaio, M.U., Correia, M.T.S., and Oliva, M.L.V. (2012).Purification, primary structure and potential functions of a novel lectin from Bauhinia forficata seeds. Process Biochem. 47, 1049–1059.10.1016/j.procbio.2012.03.008Search in Google Scholar
Zhang, F., Walcott, B., Zhou, D., Gustchina, A., Lasanajak, Y., Smith, D.F., Ferreira, R.S., Correia, M.T.S., Paiva, P.M., Bovin, N.V., et al. (2013). Structural studies of the interaction of Crataeva tapia bark protein with heparin and other glycosaminoglycans. Biochemistry 52, 2148–2156.10.1021/bi400077bSearch in Google Scholar PubMed PubMed Central
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