Evaluation of partial thromboplastin time, thrombin time and prothrombin time over treated plasma using a fibrinolytic protease
DOI:
https://doi.org/10.33448/rsd-v11i2.25439Keywords:
Thrombosis; Fibrinolytic agents; Coagulation; Plasma.Abstract
Fibrinolytic proteases are enzymes that degrade the fibrin, a main component of the blood clot. Thus, the present study aimed to understand the evaluation of aPTT, TT and PT on plasma treated with a fibrinolytic protease produced by Mucor subtillissimus. To assess the anticoagulant effect, the protease was used in different concentrations of 0.5-2.5mg/mL. The tests showed that the enzyme promoted a significant prolonged time over the PT clotting time as concentration increased. In the aPTT assay, fibrinolytic protease practically did not prolong the clotting time, even with increased enzyme concentration. In TT it was verified that at all times of contact with thrombin, be it 5-30 minutes, there was no interference on the action of thrombin or fibrinogen on the formation of thrombi. Since only the prothrombin time among all those studied was altered, it is suggested that the fibrinolytic protease under study affects the extrinsic pathway of coagulation. Thus, it was possible to observe that the serine protease promoted a significant prolonged time on the clotting time of the PT as the enzyme concentration increased, but the serine protease did not cause any type of change in TT and aPTT.
References
Banerjee, S., Prasanna, R., Bagchi, S.N. (2013). Purification and Characterization of a Fibrino(geno)lytic Protease from Cultured Natural Isolate of a Cyanobacterium, Anabaena fertilissima. Journal of Applied Social Psychology, 25, 1111- 1122.
Byskov, K., Gall, S.M.L.E., Thiede, B., Camerer, E., Kanse, S.M. (2020). Protease activated receptors (PAR)‐1 and ‐2 mediate cellular effects of factor VII activating protease (FSAP). The Faseb Journal, 34 (1), 1079-1090.
Chandramohan, M., Yee, C.Y., Beatrice, P.H.K., Ponnaiah, P., Narendrakumar, G., Samrot, A.V. (2019). Production, characterization and optimization of fibrinolytic protease from Bacillus pseudomycoides strain MA02 isolated from poultry slaughter house soils. Biocatalysis and Agricultural Biotechnology, 22, 101371.
Chang, C.T., Wang, P.M., Hung, Y.F., Chung, Y.C. (2012). Purification and biochemical properties of a fibrinolytic enzyme from Bacillus subtilis-fermented red bean. Food Chemistry, 133 (4), 1611-1617.
Devaraj, Y., Rajender, S.K., Halami, P.M. (2018). Purification and characterization of fibrinolytic protease from Bacillus amyloliquefaciens MCC2606 and analysis of fibrin degradation product by MS/MS. Preparative Biochemistry & Biotechnology, 7(48),172-180.
Fleury. (2020). Medicina e Saúde. Manual de Diagnósticos: Investigação Diagnóstica dos Distúrbios Hemorrágicos.
Gogoi, D., Arora, N., Kalita, B., Sarma, R., Islam, T., Ghosh, S.S., Devi, R., Mukherjee, A.K. (2018). Anticoagulant mechanism, pharmacological activity, and assessment of preclinical safety of a novel fibrin(ogen)olytic serine protease from leaves of Leucas indica. Scientific Reports, 8 (1), 1-17.
Hu, Y., Yu, D., Wang, Z., Hou, J., Tyagi, R., Liang, Y., Hu, Y. (2019). Purification and characterization of a novel, highly potent fibrinolytic enzyme from Bacillus subtilis DC27 screened from Douchi, a traditional Chinese fermented soybean food. Scientific Reports, 9, 9235.
Kasvi. (2019). Análise da Hemostasia: Tempo de Atividade da Protrombina (TAP) e Tempo de Tromboplastina Parcial Ativada (TTPa). https://kasvi.com.br/analise-da-hemostasia/
Kotb E. (2012). Fibrinolytic Bacterial Enzymes with Thrombolytic Activity. Egypt: Springer Briefs in Microbiology.
Kumar, S.S & Sabu A. Fibrinolytic Enzymes for Thrombolytic Therapy. In: LABROU, Nikolaos (ed.). Therapeutic Enzymes: Function and Clinical Implications. Athens: Springer, 345-381.
Lopez-Sendon J., De Lopez, S.E., Bobadilla, J.F., Rubio, R., Bermejo, J., Delcan, J.L (1995). Cardiovascular pharmacology (XIII). The efficacy of different thrombolytic drugs in the treatment of acute myocardial infarct. Revista Española de Cardiología, 48, 407–439.
Medeiros, G.M.S., Marques D.A.V., Porto, T.S., Lima-Filho, J.L., Teixeira, J.A.C., Pessoa-Júnior, A., Porto, A.L.F. (2013). Extraction of Fibrinolytic Proteases from Streptomyces sp. DPUA1576 using PEG-Phosphate Aqueous Two-Phase Systems. Fluid Phase Equilibria, 339, 52-57.
Nascimento, T.P., Sales, A.E., Porto, C.S., Brandão, R.M.P., Campos-Takaki, G.M., Teixeira, J.A.C., Porto, T.S., Porto, A.L.F., Converti, A. (2016). Purification of a fibrinolytic protease from Mucor subtilissimus UCP 1262 by aqueous two-phase systems (PEG/sulfate). Journal of Chromatography B, 16-24.
Nascimento, T.P., Sales, A.E., Porto, C.S., Brandão, R.M.P., Takaki, G.M.C., Teixeira, J.A.C., Porto, T.S., Porto, A.L.F. (2015). Production and Characterization of New Fibrinolytic Protease from Mucor subtillissimus UCP 1262 in Solid-State Fermentation. Advances in Enzyme Research, 3, 81-91.
Nascimento, T.P., Sales. A.E., Porto, T.S., Costa, R.M.P.B., Breydo, L., Uversky, V.N., Porto, A.L.F., Converti, A. (2017). Purification, biochemical, and structural characterization of a novel fibrinolytic enzyme from Mucor subtilissimus UCP 1262. Bioprocess and Biosystems Engineering, 40 (8), 1209-1219.
Opas (2017). https://www.paho.org/pt/topicos/doencas-cardiovasculares
Park, J.W., Park, J.E., Choi, H.K., Jung, T.W., Yoon, S.M., Lee, J.S. (2013). Purification and characterization of three thermostable alkaline fibrinolytic serine proteases from the polychaete Cirriformia tentaculata. Process Biochemistry, 48(10), 979-987.
Ravikumar G, Gomathi D, Kalaiselvi MUC. (2012). A protease from the medicinal mushroom Pleurotus sajor-caju; production, purification and partial characterization. Asian Pacific Journal of Tropical Biomedicine Coimbatore, 411-417.
SBC, Sociedade Brasileira de Cardiologia. 2020. Cardiômetro. http://www.cardiometro.com.br/
Silva, M.M., Rocha, T.A., Moura, D.F., Chagas, C.A., Aguiar-Júnior, F.C.A., Santos, N.P.S., Sobral, R.V.S., Nascimento, J.M., Leite, A.C.L., Pastrana, L., Brandão, R.M.P., Nascimento, T.P., Porto, A.L.F. (2019). Effect of acute exposure in swiss mice (Mus musculus) to a fibrinolytic protease produced by Mucor subtilissimus UCP 1262: An histomorphometric, genotoxic and cytological approach. Regulatory Toxicology and Pharmacology, 9(103), 282-291.
Vijayaraghavan, P., Arasu, M.V., Rajan, R.A., Al-Dhabi, N.A. (2019). Enhanced production of fibrinolytic enzyme by a new Xanthomonas oryzae IND3 using low-cost culture medium by response surface methodology. Saudi Journal of Biological Sciences, 26, 217-224.
Wang, S.L., Wu, Y.Y., Liang, T.W. (2011). Purification and biochemical characterization of a nattokinase by conversion of shrimp shell with Bacillus subtilis TKU007. New Biotechnology, 28(2), 196-202.
Zago, M.A., Falcão, R.P., Pasquini, R., Spector, N., Covas, D.T., Rego E.M. (2013). Tratado de Hematologia. São Paulo: Atheneu.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2022 Vanessa Maria Andrade Machado Miranda; José Pedro Martins Barbosa Filho; Romero Marcos Pedrosa Brandão Costa; Ana Cristina Lima Leite; Vagne de Melo Oliveira; Juanize Matias da Silva Batista; Lorenzo Pastrana; Thiago Pajeú Nascimento; Ana Lúcia Figueiredo Porto
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
1) Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
2) Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
3) Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.
