In vitro inhibition of thrombin generation by sulfated polysaccharides from the tropical red seaweed Gracilaria birdiae Plastino & Oliveira / Inibição in vitro da geração de trombina por polissacarídeos sulfatados da alga marinha vermelha tropical Gracilaria birdiae Plastino & Oliveira

Autores

  • José Ariévilo Gurgel Rodrigues Universidade Federal do Rio de Janeiro
  • Priscila Gomes Barcellos UFRJ
  • Tayane Caetana Salles UFRJ
  • Norma Maria Barros Benevides UFC
  • Ana Maria Freire Tovar UFRJ
  • Paulo Antônio de Souza Mourão UFRJ

DOI:

https://doi.org/10.2312/Actafish.2017.5.1.22-32

Palavras-chave:

Rhodophyta, marine environment, polysulfated, chemical analysis, thrombin.

Resumo

Abstract Gracilaria birdiae is a red macroalga found on the Brazilian coastline and experimentally cultivated to obtain industrially-important sulfated polysaccharides (SPs) for biotechnology, but their anticoagulant potentials on thrombin generation (TG) in vitro have not been examined. This study compared the in vitro inhibition of SPs isolated and physical-chemically characterized from samples of G. birdiae collected in natural-bed (Gbnb) and cultured (Gbc) on the coastal of Flecheiras beach on TG. Papain extraction yield difference reached 11.76%, and different DEAE-cellulose chromatograms were obtained, with fractions (Gbnb-I→II and Gbc-I→IV) revealing charge density patterns and molecular sizes > 100 kDa according to the both agarose and polyacrylamide electrophoreses, respectively. These procedures, associated with toluidine blue/Stains-All staining, also revealed nonSPs. By both activated partial thromboplastin time (APTT) and prothrombin time assays, fractions had no anticoagulant actions, except Gbc-I that discretely altered the APTT (0.51 IU mg mL-1) compared with unfractionated heparin (UHEP, 193 IU mg mL-1). Gbc-I inhibited concentration-dependent both intrinsic and extrinsic coagulation pathways in in vitro TG assay using 60-fold diluted human plasma, with 50% efficacy on thromboplastin-activated TG, similar to Gbnb-I, although less potent than UHEP which abolished it at a 41.65-fold lower concentration. The results suggest that G. birdiae change its matrix-polysaccharide content under field conditions and TG assay could guide complementary analyses on algal species anticoagulant potential for pharmacological use.

Biografia do Autor

José Ariévilo Gurgel Rodrigues, Universidade Federal do Rio de Janeiro

Graduado em Engenharia de Pesca pela Universidade Federal do Ceará (2004), mestrado em Recursos Pesqueiros e Engenharia de Pesca pela Universidade Federal do Ceará (2006) e doutorado em Biotecnologia pela Rede Nordeste de Biotecnologia - RENORBIO/UFC (2011). Tem experiência na área de Recursos Pesqueiros e Engenharia de Pesca e de Biotecnologia em Recursos Naturais, com ênfase em Taxonomia de Algas Marinhas, Fisiologia Comparada (taxonomia de grupos, filogenia e biogeografia) e Bioprospecçâo de Produtos Naturais Bioativos de Organismos Aquáticos. Atua nas seguintes áreas: Ficologia, Química de macromoléculas (polissacarídeos sulfatados) e suas propriedades químicas e implicações terapêuticas sobre coagulação, trombose, dor, inflamação e de avaliação toxicológica (aguda, subcrônica e crônica). Foi professor horista (prestador de serviços) do Instituto de Desenvolvimento, Educação e Cultura do Ceará (IDECC-UVA) e do Instituto Dom José de Educação e Cultura (IDJ-UVA). Atualmente, realiza Pós-Doutorado na Universidade Federal do Rio de Janeiro (CAPES-PNPD).

Referências

Amorim, R.N.S., Rodrigues, J.A.G., Holanda, M.L., Quinderé, A.L.G., Paula, R.C.M., Melo, V.M.M. & Benevides, N.M.B. (2012). Antimicrobial effect of a crude sulfated polysaccharide from the red seaweed Gracilaria ornata. Braz. Arch. Biol. Technol., 55(2): 171-181.

Athukorala,Y., Jung, W.K., Vasanthan, T. & Jeon, Y.J. (2006). An anticoagulative polysaccharide from an enzymatic hydrolysate of Ecklonia cava. Carbohydr. Polym., 66(2): 184-191.

Bezerra, A.F. & Marinho-Soriano, E. (2010). Cultivation of the red seaweed Gracilaria birdiae (Gracilariales, Rhodophyta) in tropical waters of northeast Brazil. Biom. Bio., 34(12): 1813-1817.

Chang, C.W., Lur, H.S., Lu, M.K. & Cheng, J.J. (2013). Sulfated polysaccharides of Amillariella mellea and their anti-inflammatory activities via NF-κB suppression. Food Res. Inter., 54(1): 239-245.

Costa-Lotufo, L.V., Pessoa, C., Moraes, M.E.A., Almeida, A.M.P., Moraes, M.O. & Lotufo, T.M.C. Marine organisms from Brazil as source of potential anticancer agents. Adv. Phytom., 2(1): 181-196.

Castoldi, E. & Rosing J. (2011). Thrombin generation tests. Throm. Res., 127(suppl.3): S21-S25.

Dantas-Santos, N., Gomes, D.L., Costa, L.S., Cordeiro, S.L., Costa, M.S.S.P., Trindade, E.S., Chavichiolo, C.R., Scortecci, K.C., Leite, E.L. & Rocha, H.A.O. (2012). Freschwater plants synthesize sulfated polysaccharides: Heterogalactans from water hycinth (Eicchornia crassipes). Inter. J. Mol. Sci., 13(1): 961-976.

Dietrich, C.P.; & Dietrich, S.M.C. (1976). Electrophoretic behaviour of acidic mucopolysaccharides in diamine buffers. Anal. Biochem.,70(2): 645-647.

Farndale, R.W., Buttle, D.J. & Barrett, A.J. (1986). Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochim. Biophys. Acta, 883(2): 173-177.

Fidelis, G.P., Camara, R.B.G., Queiroz, M.F., Costa, M.S.S.P., Santos, P.C., Rocha, H.A.O. & Costa, L. S. (2014). Proteolysis, NaOH and ultrasound-enhanced extraction of anticoagulant and antioxidant sulfated polysaccharides from the edible seaweed, Gracilaria birdiae. Mol., 19(11), 18511-18526.

Glauser, B.F., Rezende, R.M., Melo, F.R., Pereira, M.S., Fransischetti, I.M.B., Monteiro, R.Q., Rezaie, A.R. & Mourão, P.A.S. (2009). Anticoagulant activity of a sulfated: Serpin-independent effect and specific interaction with factor Xa. Throm. Haem., 102(6): 1183-1193.

Hung, L.D., Hori, K., Nang, H.Q., Kha, T. & Hoa, L.T.(2009). Seasonal changes in growth rate, carrageenan yield and lectin content in the red alga Kappaphycus alvarezii cultivated in Camranh Bay, Vietnam. J. Appl.Phycol., 21(3): 265-272.

Maciel, J.S., Chaves, L.S., Souza, B.W.S., Teixeira, D.I.A., Freitas, A.L.P., Feitosa, J.P.A. & Paula, R. C.M. (2008). Structural characterization of cold extracted fraction of soluble sulfated polysaccharide from red seaweed Gracilaria birdiae. Carbohydr. Polym., 71(4): 55-565.

Mourão, P.A.S. (2015). Perspective on the use of sulfated polysaccharides from marine organisms as a source of new antithrombotic drugs. Mar. Drugs, 1(5): 2770-2784.

Mourão, P.A.S., Boisson-Vidal, C., Tapon-Bretaudière, J., Drouet, B., Bros, A. & Fischer, A.M. (2001). Inactivation of thrombin by a fucosylated chondroitin sulfate from Echinoderm. Throm. Res., 102(2): 167-176.

Nishino, T., Fukuda, A., Nagumo, T., Fujihara, M. & Kaji, E. (1999). Inhibition of the generation of thrombin and factor Xa by a fucoidan from the brown seaweed Ecklonia kurome. Throm. Res., 96(1): 37-49.

Pereira, R.C. & Costa-Lotufo, L.V. (2012). Bioprospecting for bioactives from seaweeds: potential, obstacles and alternatives. Braz. J. Pharmacog., 22(4): 894-905.

Pomin, V.H. (2012). Fucanomis and galactanomics: Current status in drug discovery, mechanisms of action and role of the well-defined structures. Biochim. Biophys. Acta, 1820(12): 1971-1979.

Prajapati, V.D., Maheriya, P.M., Jani, G.K. & Solanki, H.K. (2014). Carrageenan: A natural seaweed polysaccharide and its applications. Carbohydr. Polym., 105(25): 97-112.

Rau, J.C., Beaulieu, L.M., Huntington, J.A. & Church, F.C. Serpins in thrombosis, hemostasis and fibrinlysis. J. Throm. Haem., 5(suppl.1): 102-115.

Rodrigues, J.A.G., Araújo, I.W.F., Paula, G.A., Lima, T.B., Bessa, E.F. & Benevides, N.M.B. (2011). Carragenana da epífita Hypnea musciformis obtida do cultivo experimental de Solieria filiformis em Flecheiras, Estado do Ceará, Brasil. Acta Sci. Technol., 33(2): 137-144.

Rodrigues, J.A.G., Neto, E.M., Teixeira, L.A.C., Paula, R.C.M., Mourão, P.A.S. & Benevides, N.M.B. (2013). Structural features and inactivation of coagulation proteases of a sulfated polysaccharidic fraction from Caulerpa cupressoides var. lycopodium (Caulerpaceae, Chlorophyta). Acta Sci. Technol., 35(4): 611-619.

Rodrigues, J.A.G., Queiroz, I.N.L., Quinderé, A.L.G., Benevides, N.M.B., Tovar, A.M.F., & Mourão, P.A.S. (2016). Mild-acid hydrolysis of a native polysulfated fraction from Acanthophora muscoides generates sulfated oligosaccharides displaying in vitro thrombin generation inhibition. Acta Sci. Biol. Sci., 38(1): 7-15.

Souza, B.W.S., Cerqueira, M.A., Bourbon, A.I., Pinheiro, A.C., Martins, J.T., Teixeira, J.A., Coimbra, M.A. & Vicente, A.A. (2012). Chemical characterization and antioxidant activity of sulfated polysaccharide from the red seaweed Gracilaria birdiae. Food Hydrocoll., 27(2): 287-292.

Vanderlei, E.S.O., Araújo, I.W.F., Quinderé, A.L.G., Fontes, B.P., Eloy, Y.R.G., Rodrigues, J.A.G., Silva, A.A.R., Chaves, H.V., Jorge, R.J.B., Menezes, D.B., Evangelista, J.S.A.M., Bezerra, M.M. & Benevides, N.M.B. (2011). The involvement of the HO-1 pathway in the anti-inflammatory action of a sulfated polysaccharide isolated from the red seaweed Gracilaria birdiae. Inflamm. Res., 6(12): 1121-1130.

Volpi, N. & Maccari, F. (2002). Detection of submicrogram quantities of glycosaminoglycans on agarose gels by sequential staining with toluidin blue and Stains-All. Electrophor., 23(24): 4060-4066.

Zhang, Z., Till, S., Jiang, C., Knappe, S., Reutterer, S., Scheiflinger, F., Szabo, C.M. & Dockai, M. (2014). Structure-activity relationship of the pro- and anticoagulant effects of Fucus vesiculosus fucoidan. Throm. Haem., 111(3): 429-437.

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Publicado

2018-05-01