Planta Med 2008; 74(7): 712-718
DOI: 10.1055/s-2008-1074522
Pharmacology
Original Paper
© Georg Thieme Verlag KG Stuttgart · New York

A Non-Anticoagulant Heterofucan has Antithrombotic Activity in vivo

Edjane M. A. Barroso1 , Leandro S. Costa1 , Valquíria P. Medeiros2 , Sara L. Cordeiro1 , Mariana S. S. P. Costa1 , Célia R. C. Franco3 , Helena B. Nader2 , Edda L. Leite1 , Hugo A. O. Rocha1
  • 1Programa de Pós-graduação em Ciências da Saúde, Departamento de Bioquímica, Universidade Federal do Rio Grande do Norte - UFRN, Natal - RN, Brazil
  • 2Departamento de Bioquímica, Universidade Federal de São Paulo - UNIFESP, São Paulo - SP, Brazil
  • 3Departamento de Biologia Celular, Universidade Federal do Paraná - UFPR, Curitiba - PR, Brazil
Further Information

Publication History

Received: September 7, 2007 Revised: March 9, 2008

Accepted: March 11, 2008

Publication Date:
21 May 2008 (online)

Abstract

Fucan is a term used to denominate a family of sulfated L-fucose-rich polysaccharides. The brown alga Spatoglossum schröederi (Dictyotaceae) has three heterofucans namely fucan A, B and C. The 21 kDa fucan A is composed of a core of a β (1 - 3) glucuronic acid-containing oligosaccharide of 4.5 kDa with branches at C4 of the fucose chains α (1 - 3) linked. The fucose is mostly substituted at C4 with a sulfate group and at C2 with chains of β (1 - 4) xylose. This fucan has neither anticoagulant (from from 0.1 to 100 μg) nor hemorrhagic activities (from 50 to 800 μg/mL). The antithrombotic test in vivo showed that fucan A has no activity in any of the concentrations (from 0.2 to 20 μg/g/day) tested 1 h after polysaccharide administration. However, when fucan A was injected endovenously 24 h before the ligature of the venae cavae, we observed a dose-dependent effect, reaching saturation at around 20 μg/g of rat weight. In addition, this effect is also time-dependent, reaching saturation around 16 h after fucan administration. In addition, regardless of the administration route, fucan A displayed antithrombotic activity. The exception was the oral pathway. Of particular importance was the finding that fucan A stimulates the synthesis of an antithrombotic heparan sulfate from endothelial cells like heparin. The hypothesis has been raised that the in vivo antithrombotic activity of fucan A is related to the increased production of this heparan. Taken together with the fact that the compound is practically devoid of anticoagulant and hemorrhagic activity, the data suggest that it may be an ideal antithrombotic agent in vivo.

Abbreviations

APTT: activated partial thromboplastin time

CS: chondroitin sulfate

FUC: fucoidan from Fucus vesiculosus

FUC A: fucan A from Spatoglossum schröederi

HEP: heparin

HS: heparin sulfate

PRP: platelet-rich plasma

PPP: platelet-poor plasma

PT: protrombin time

TT: thrombin time

References

  • 1 Nader H B, Lopes C C, Rocha H AO, Santos E A, Dietrich C P. Heparins and heparinoids: Occurrence, structure and mechanism of antithrombotic and hemorrhagic activities.  Curr Pharm Design. 2004;  10 951-66
  • 2 Liu J, Pedersen L C. Anticoagulant heparan sulfate: structural specificity and biosynthesis.  Appl Microbiol Biotechnol. 2007;  74 263-72
  • 3 Nader H B, Buonassisi V, Colburn P, Dietrich C P. Heparin stimulates the synthesis and modifies the sulfation pattern of heparan-sulfate proteoglycan from endothelial cells.  J Cell Physiol. 1989;  140 305-10
  • 4 Berteau O, Mulloy B. Sulfated fucans, fresh perspectives: structures, functions, and biological properties of sulfated fucans and an overview of enzymes active toward this class of polysaccharide.  Glycobiology. 2003;  13 29R-40R
  • 5 Vilela-Silva A, Castro M O, Valente A P, Biermann C H, Mourao P AS. Sulfated fucans from the egg jellies of the closely related sea urchins Strongylocentrotus droebachiensis and Strongylocentrotus pallidus ensure species-specific fertilization.  J Biol Chem. 2002;  277 379-87
  • 6 Rocha H AO, Leite E L, Medeiros V P, Lopes C C, Nascimento F D, Tersariol I LS. et al . Natural sulfated polysaccharides as antithrombotic compounds. Structural characteristcs and effects on the coagulation cascade. In: Verli H, editor. Insights into carbohydrate structure and biological function.  Kerala: Transworld Research. Network;  2006 51-67
  • 7 Millet J, Jouault S C, Mauray S, Theveniaux J, Sternberg C, Boisson Vidal C. et al . Antithrombotic and anticoagulant activities of a low molecular weight fucoidan by the subcutaneous route.  Thromb Haemostasis. 1999;  81 391-5
  • 8 Mauray S, Sternberg C, Theveniaux J. et al . Venous antithrombotic and anticoagulant activities of a fucoïdan fraction.  Thromb Haemostasis. 1995;  74 1280-5
  • 9 Soeda S, Ohmagari Y, Shimeno H, Nagamatsu A. Preparation of aminated fucoidan and its evaluation as an antithrombotic and antilipemic agent.  Biol Pharm Bull. 1994;  17 784-8
  • 10 Leite E L, Medeiros M GL, Rocha H AO, Farias G GM, da Silva L F, Chavante S F. et al . Structure and pharmacological activities of a sulfated xylofucoglucuronan from the alga Spatoglossum schroederi. .  Plant Sci. 1998;  132 215-28
  • 11 Rocha H AO, Bezerra L C, de Albuquerque I RL, Costa L S, Guerra C MP, de Abreu L RD. et al . A xylogalactofucan from the brown seaweed Spatoglossum schröederi stimulates the synthesis of an antithrombotic heparan sulfate from endothelial cells.  Planta Med. 2005;  71 379-81
  • 12 Rocha H AO, Moraes F A, Trindade E S, Franco C RC, Torquato R JS, Veiga S S. et al . Structural and hemostatic activities of a sulfated galactofucan from the brown alga Spatoglossum schröederi - An ideal antithrombotic agent?.  J Biol Chem. 2005;  280 41 278-88
  • 13 Buonassisi V, Venter J C. Hormone and neurotransmiter receptors in an established vascular endothelial cell line.  Proc Natl Acad Sci U S A. 1976;  73 612-6
  • 14 Nader H B, Chavante S F, Santos E A, Oliveira F W, Paiva J F, Jeronimo S M B. et al . eparan sulfates and heparins: similar compounds performing the same functions in vertebrates and invertebrates?.  HBraz J Med Biol Res. 1999;  32 529-38
  • 15 Dische Z A. A new specific color reaction of hexuronic acid.  J Biol Chem. 1974;  167 189-98
  • 16 Dische Z. Color reactions of hexuronic acids, Vol. 1. London; Academic Press 1962: 479-81
  • 17 Dische Z. Color reaction of pentoses, Vol. 1. London; Academic Press 1962: 497-501
  • 18 Dubois M, Gilles K A, Hamilton J K, Rebers P A, Smith F. Colorimetric method for determination of sugars and related substances.  Anal Chem. 1956;  28 350-6
  • 19 Nader H B, Dietrich C P. Determination of sulfate after chromatography and toluidine blue complex-formation.  Anal Biochem. 1977;  78 112-8
  • 20 Spector J. Refinement of the coomassie blue method of protein quantification. A simple and linear spectrophotometric assay of 0.5 to 50 μg of protein.  Anal Biochem. 1978;  86 142-3
  • 21 Taylor R L, Conrad H E. Stoichiometric depolymerization of polyuronides and glycosaminoglycuronans to monosaccharides following reduction of their carbodiimide-activated carboxyl groups.  Biochemistry. 1972;  11 1383-8
  • 22 Dietrich C P, Shinjo S T, Moraes F A, Castro R AB, Mendes A, Gouvea T C. et al . Structural features and bleeding activity of commercial low molecular weight heparins: Neutralization by ATP and protamine.  Semin Thromb Hemostasis. 1999;  25 43-50
  • 23 Nader H B, Dietrich C P, Buonassisi V, Colburn P. Heparin sequences in the heparan-sulfate chains of an endothelial-cell proteoglycan.  Proc Natl Acad Sci U S A. 1987;  84 3565-9
  • 24 Rocha H AO, Franco C R, Trindade E S, Carvalho L C, Veiga S S, Leite E L. et al . A fucan from the brown seaweed Spatoglossum schröederi inhibits Chinese hamster ovary cell adhesion to several extracellular matrix proteins.  Braz J Med Biol Res. 2001;  34 621-6
  • 25 Farias W RL, Valente A P, Pereira M S, Mourão P AS. Structure and anticoagulant activity of sulfated galactans. Isolation of a unique sulfated galactan from the red algae Botryocladia occidentalis and comparison of its anticoagulant action with that of sulfated galactans from invertebrates.  J Biol Chem. 2000;  38 29 299-307
  • 26 Buonassisi V, Colburn P. Biological significance of heparan-sulfate proteoglycans.  Ann N Y Acad Sci. 1982;  401 6-84
  • 27 Glabe C G, Harty P K, Rosen S D. Preparation and properties of fluorescent polysaccharides.  Anal Biochem. 1983;  130 287-94

Hugo Alexandre de Oliveira Rocha

Universidade Federal do Rio Grande do Norte

Centro de Biociências

Departamento de Bioquímica

Laboratório de Biotecnologia de Polímeros Naturais-BIOPOL

Av. Salgado Filho S/N

Natal - RN

Brazil

Phone: +55-84-3211-9208

Email: hugo@cb.ufrn.br