Unique extracellular matrix heparan sulfate from the bivalve Nodipecten nodosus (Linnaeus, 1758) safely inhibits arterial thrombosis after photochemically induced endothelial lesion.
Heparin-like glycans with diverse disaccharide composition and high anticoagulant activity have been described in several families of marine mollusks. The present work focused on the structural characterization of a new heparan sulfate (HS)-like polymer isolated from the mollusk Nodipecten nodosus (Linnaeus, 1758) and on its anticoagulant and antithrombotic properties. Total glycans were extracted from the mollusk and fractionated by ethanol precipitation. The main component (>90%) was identified as HS-like glycosaminoglycan, representing approximately 4.6 mg g(-1) of dry tissue. The mollusk HS resists degradation with heparinase I but is cleaved by nitrous acid. Analysis of the mollusk glycan by one-dimensional (1)H, two-dimensional correlated spectroscopy, and heteronuclear single quantum coherence nuclear magnetic resonance revealed characteristic signals of glucuronic acid and glucosamine residues. Signals corresponding to anomeric protons of nonsulfated, 3- or 2-sulfated glucuronic acid as well as N-sulfated and/or 6-sulfated glucosamine were also observed. The mollusk HS has an anticoagulant activity of 36 IU mg(-1), 5-fold lower than porcine heparin (180 IU mg(-1)), as measured by the activated partial thromboplastin time assay. It also inhibits factor Xa (IC(50) = 0.835 microg ml(-1)) and thrombin (IC(50) = 9.3 microg ml(-1)) in the presence of antithrombin. In vivo assays demonstrated that at the dose of 1 mg kg(-1), the mollusk HS inhibited thrombus growth in photochemically injured arteries. No bleeding effect, factor XIIa-mediated kallikrein activity, or toxic effect on fibroblast cells was induced by the invertebrate HS at the antithrombotic dose.
Gomes
Angélica M
AM
Laboratório de Tecido Conjuntivo, Instituto de Bioquímica Médica, Programa de Glicobiologia and Hospital Universitário Clementino Fraga Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro CEP 21941-590, Brasil.
Kozlowski
Eliene O
EO
Pomin
Vitor H
VH
de Barros
Cintia Monteiro
CM
Zaganeli
José L
JL
Pavão
Mauro S G
MS
D000818:Animals; D000925:Anticoagulants / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism* / Q000627:therapeutic use; D000991:Antithrombins / Q000378:metabolism; D001158:Arteries / Q000187:drug effects / Q000473:pathology / Q000528:radiation effects; D049872:Bivalvia / Q000378:metabolism; D002236:Carbohydrate Conformation; D002341:Carotid Artery Thrombosis / Q000517:prevention & control*; D002460:Cell Line; D004730:Endothelium, Vascular / Q000187:drug effects / Q000473:pathology / Q000528:radiation effects; D005109:Extracellular Matrix / Q000737:chemistry* / Q000378:metabolism; D015951:Factor Xa / Q000378:metabolism; D065427:Factor Xa Inhibitors; D005260:Female; D019812:Heparan Sulfate Proteoglycans / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism* / Q000627:therapeutic use*; D006493:Heparin / Q000378:metabolism / Q000627:therapeutic use; D015844:Heparin Cofactor II / Q000378:metabolism; D019763:Heparin Lyase / Q000378:metabolism; D006801:Humans; D008297:Male; D009608:Nitrous Acid / Q000378:metabolism; D051381:Rats; D013057:Spectrum Analysis / Q000379:methods; D013552:Swine; D013917:Thrombin / Q000037:antagonists & inhibitors / Q000378:metabolism
D000925:Anticoagulants; D000991:Antithrombins; D065427:Factor Xa Inhibitors; D019812:Heparan Sulfate Proteoglycans; D015844:Heparin Cofactor II; D006493:Heparin; D013917:Thrombin; D015951:Factor Xa; D019763:Heparin Lyase; D009608:Nitrous Acid