Extraction, physical-chemical characterization and in vitro inhibitory potential of thrombin generation of crude sulfated polysaccharides from Brazilian tropical seaweeds
Document title: Extraction, physical-chemical characterization and in vitro inhibitory potential of thrombin generation of crude sulfated polysaccharides from Brazilian tropical seaweeds
Journal: Acta scientiarum. Biological sciences
Database: PERIÓDICA
System number: 000433637
ISSN: 1679-9283
Authors: 1
1
1
2
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Institutions: 1Universidade Federal do Rio de Janeiro, Hospital Universitario Clementino Fraga Filho, Rio de Janeiro. Brasil
2Universidade Federal do Ceara, Departamento de Bioquimica e Biologia Molecular, Fortaleza, Ceara. Brasil
Year:
Volumen: 40
Country: Brasil
Language: Inglés
Document type: Artículo
Approach: Analítico, descriptivo
English abstract The biotechnological value of macroalgae for screening assays of thrombin generation-TG using sulfated polysaccharides-SPs as substitutes to heparin has been poorly explored. Five Brazilian species of macroalgae (Gracilaria birdiae, Acanthophora muscoides, Halymenia sp., Caulerpa cupressoides and C. racemosa) were analyzed and compared for their abundance, physical-chemical characteristics and in vitro anticoagulant assays of activated partial thromboplastin time-APTT, prothrombin time-PT and TG. Papain extraction yielded (p < 0.001) from 0.66±0.03% (C. racemosa) to 41.60±1.10% (Halymenia sp.) of crude SPs varying sulfate (8.41-42.60%) and total sugars (47.80-70.53%). Crude SPs showed difference in mobility and resolution pattern by agarose electrophoresis, while polyacrylamide analysis revealed SPs of > 100 kDa. These procedures, combined with the use of Stains-All, also indicated nonSPs. APTTs ranged from 2.81 (A. muscoides) to 21.30 IU (Halymenia sp.) vs. heparin (193 IU), and were dependent on sulfation of the crude SPs. PT was not altered. With respect to TG assay, crude SPs modified concentration-dependent and independently from molecular mass TG by both intrinsic/extrinsic pathways in 60-fold diluted human plasma, with total intrinsic inactivation using crude SPs from A. muscoides in parallel to heparin (p < 0.05). Thrombosis in vitro is differentially modulated by distinct crude SPs from Brazilian seaweeds
Portuguese abstract O valor biotecnológico das macroalgas para ensaios de varredura de geração de trombina-GT pouco tem sido explorado usando polissacarídeos sulfatados-PSs como substitutos à heparina. Foram analisadas e comparadas cinco espécies brasileiras de macroalgas (Gracilaria birdiae, Acanthophora muscoides, Halymenia sp., Caulerpa cupressoides e C. racemosa) quanto à abundância, às características físico-químicas e os ensaios anticoagulantes in vitro de tempo de tromboplastina parcial ativada-TTPA, ao tempo de protrombina-TP e a GT. A extração com papaína rendeu (p < 0,001) de 0,66±0,03% (C. racemosa) a 41,60±1,10% (Halymenia sp.) de PSs brutos variando sulfato (8,41-42,60%) e açúcares totais (47,80-70,53%). Os PSs brutos mostraram diferenças na mobilidade e resolução por eletroforese em agarose, enquanto pela análise em poliacrilamida revelou PSs brutos de >100 kDa. Esses procedimentos, combinados ao uso de azul de toluidina/Stains-All, indicaram também polissacarídeos-não sulfatados. Os TTPAs foram dependentes da sulfatação dos PSs brutos e variaram de 2,81 (A. muscoides) a 21,30 UI (Halymenia sp.) vs. heparina (193 UI). O TP não foi alterado. Com respeito ao ensaio de GT, os PSs brutos modificaram, dependente de concentração e independentemente de massa molecular, GT pelas vias intrínseca/extrínseca no plasma humano diluído 60 vezes, com inativação intrínseca total usando PSs brutos de A. muscoides em paralelo à heparina (p < 0,05). A trombose in vitro é modulada diferencialmente por PSs brutos distintos de algas marinhas brasileiras
Disciplines: Biología,
Medicina
Keyword: Algas,
Fitoquímica,
Hematología,
Diagnóstico,
Algas marinas,
Polisacáridos sulfatados,
Glucanos,
Pruebas de coagulación,
Trombina
Keyword: Algae,
Phytochemistry,
Hematology,
Diagnosis,
Marine algae,
Sulfated polysaccharides,
Glycans,
Coagulation tests,
Thrombin
Full text: http://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/34463/pdf