Homogalactanas sulfatadas da alga Codium isthmocladum com atividade anticoagulante
Since the first description of sulfated polysaccharides from seaweeds, the biological activities of these compounds have been evaluated under different aspects and experimental procedures. Among the broad biological activities presented by seaweed polysaccharides, anticoagulant action appears as a p...
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Alga - (Codium isthmocladum) Atividade anticoagulante Galactanas sulfatadas Codium isthmocladum Anticoagulant activity Sulfated galactans CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA |
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Alga - (Codium isthmocladum) Atividade anticoagulante Galactanas sulfatadas Codium isthmocladum Anticoagulant activity Sulfated galactans CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA Farias, Eduardo Henrique Cunha de Homogalactanas sulfatadas da alga Codium isthmocladum com atividade anticoagulante |
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Since the first description of sulfated polysaccharides from seaweeds, the biological activities of these compounds have been evaluated under different aspects and experimental procedures. Among the broad biological activities presented by seaweed polysaccharides, anticoagulant action appears as a promising function. In this present study we have obtained sulfated polysaccharides from the green seaweed Codium isthmocladium by proteolytic digestion, followed by separation into five fractions (0.3, 0.5, 0.7, 0.9 and 1.2) by sequential acetone precipitation. The chemical analyses have demonstrated that all fractions are composed mainly by sulfated polysaccharides. The anticoagulant activity of these fractions was determined by activated partial thromboplastin time (aPTT) and prothrombin time test (PT) using citrate normal human plasma. None fraction has shown anticoagulant activity by PT test. Furthermore, all of them have shown anticoagulant activity by aPTT test. These results indicated that the molecular targets of these sulfated polysaccharides are mainly in the intrinsic via of the coagulation cascade. Agarose gel electrophoresis in 1,3-diaminopropane acetate buffer, pH 9.0, stained with 0.1% toluidine blue showed the presence of two or three bands in several fractions while the fraction 0.9 showed a single spot. By anion exchange chromatography, the acid polysaccharides from the 0.9 acetone fraction were separated into two new fractions eluted respectively with 2.0 and 3.0 M NaCl. These compounds showed a molecular weight of 6.4 and 7.4 kDa respectively. Chemical analyses and infrared spectroscopy showed that Gal 1 and Gal 2 are sulfated homogalactans and differ one from the other in degree and localization of sulfate groups. aPPT test demonstrated that fractions 2,0 and 3,0M (Gal1 and Gal 2, respectively) have anticoagulant activity. This is the first time that anticoagulant sulfated homogalatans have been isolated from green algae. To prolong the coagulation time to double the baseline value in the aPTT, the required amount of sulfated galactan 1 (6,3mg) was similar to low molecular heparin Clexane®, whereas only 0,7mg of sulfated galactan 2 was needed to obtain the same effect. Sulfated galactan 2 in high doses (250mg) induces platelet aggregation. These results suggest that these galactans from C. isthmocladum have a potential application as an anticoagulant drug |
| author2 |
Rocha, Hugo Alexandre de Oliveira |
| author_facet |
Rocha, Hugo Alexandre de Oliveira Farias, Eduardo Henrique Cunha de |
| format |
masterThesis |
| author |
Farias, Eduardo Henrique Cunha de |
| author_sort |
Farias, Eduardo Henrique Cunha de |
| title |
Homogalactanas sulfatadas da alga Codium isthmocladum com atividade anticoagulante |
| title_short |
Homogalactanas sulfatadas da alga Codium isthmocladum com atividade anticoagulante |
| title_full |
Homogalactanas sulfatadas da alga Codium isthmocladum com atividade anticoagulante |
| title_fullStr |
Homogalactanas sulfatadas da alga Codium isthmocladum com atividade anticoagulante |
| title_full_unstemmed |
Homogalactanas sulfatadas da alga Codium isthmocladum com atividade anticoagulante |
| title_sort |
homogalactanas sulfatadas da alga codium isthmocladum com atividade anticoagulante |
| publisher |
Universidade Federal do Rio Grande do Norte |
| publishDate |
2014 |
| url |
https://repositorio.ufrn.br/jspui/handle/123456789/12526 |
| work_keys_str_mv |
AT fariaseduardohenriquecunhade homogalactanassulfatadasdaalgacodiumisthmocladumcomatividadeanticoagulante |
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1773963259344322560 |
| spelling |
ri-123456789-125262019-01-29T19:46:59Z Homogalactanas sulfatadas da alga Codium isthmocladum com atividade anticoagulante Farias, Eduardo Henrique Cunha de Rocha, Hugo Alexandre de Oliveira http://lattes.cnpq.br/0933304924768138 http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4799567J8&dataRevisao=null Ferreira, Carmem Veríssima Leite, Edda Lisboa http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783254U6&dataRevisao=null Alga - (Codium isthmocladum) Atividade anticoagulante Galactanas sulfatadas Codium isthmocladum Anticoagulant activity Sulfated galactans CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA Since the first description of sulfated polysaccharides from seaweeds, the biological activities of these compounds have been evaluated under different aspects and experimental procedures. Among the broad biological activities presented by seaweed polysaccharides, anticoagulant action appears as a promising function. In this present study we have obtained sulfated polysaccharides from the green seaweed Codium isthmocladium by proteolytic digestion, followed by separation into five fractions (0.3, 0.5, 0.7, 0.9 and 1.2) by sequential acetone precipitation. The chemical analyses have demonstrated that all fractions are composed mainly by sulfated polysaccharides. The anticoagulant activity of these fractions was determined by activated partial thromboplastin time (aPTT) and prothrombin time test (PT) using citrate normal human plasma. None fraction has shown anticoagulant activity by PT test. Furthermore, all of them have shown anticoagulant activity by aPTT test. These results indicated that the molecular targets of these sulfated polysaccharides are mainly in the intrinsic via of the coagulation cascade. Agarose gel electrophoresis in 1,3-diaminopropane acetate buffer, pH 9.0, stained with 0.1% toluidine blue showed the presence of two or three bands in several fractions while the fraction 0.9 showed a single spot. By anion exchange chromatography, the acid polysaccharides from the 0.9 acetone fraction were separated into two new fractions eluted respectively with 2.0 and 3.0 M NaCl. These compounds showed a molecular weight of 6.4 and 7.4 kDa respectively. Chemical analyses and infrared spectroscopy showed that Gal 1 and Gal 2 are sulfated homogalactans and differ one from the other in degree and localization of sulfate groups. aPPT test demonstrated that fractions 2,0 and 3,0M (Gal1 and Gal 2, respectively) have anticoagulant activity. This is the first time that anticoagulant sulfated homogalatans have been isolated from green algae. To prolong the coagulation time to double the baseline value in the aPTT, the required amount of sulfated galactan 1 (6,3mg) was similar to low molecular heparin Clexane®, whereas only 0,7mg of sulfated galactan 2 was needed to obtain the same effect. Sulfated galactan 2 in high doses (250mg) induces platelet aggregation. These results suggest that these galactans from C. isthmocladum have a potential application as an anticoagulant drug Desde a primeira descrição de polissacarídeos sulfatados em algas marinhas, as atividades biológicas destes compostos foram avaliadas sob diferentes aspectos e procedimentos experimentais. Dentre as diversas atividades biológicas apresentadas pelos polissacarídeos de algas marinhas, a ação do anticoagulante aparece como sendo uma função promissora. Neste presente estudo foram obtidas cinco frações de polissacarídeos sulfatados da alga Codium isthmocladium (F0.3, F0.5, F0.7, F0.9 e F1.2) , através de proteólise seguida de fracionamento com volumes crescentes de acetona. Análises químicas demonstraram que todas as frações são compostas por polissacarídeos sulfatados. A atividade anticoagulante destas frações cetônicas foram determinadas por testes de tempo de tromboplastina parcial ativada (aPTT) e tempo de protrombina (PT), utilizando plasma humano normal citratado. Nenhuma fração apresentou atividade via teste de PT. No entanto todas as frações demonstraram atividade anticoagulante para teste de aPPT. Estes resultados indicam que o(s) seus alvo(s) molecular(es) esta(ão) localizado(s) principalmente na via intrínseca da cascata de coagulação. Eletroforese em gel de agarose demonstrou a presença de 2 ou 3 bandas em varias frações cetônicas, enquanto a F0.9, somente uma banda. Os polissacarídeos da F 0.9, quando submetidos a cromatografia da troca iônica, foram separados em duas novas frações eluídas com 2,0 e 3,0 M de NaCl. Estes compostos apresentaram um peso molecular de 6.4 e 7.4 kDa respectivamente. Analises químicas e espectroscopia de infravermelho das Gal 1 e Gal 2 demonstraram que essas são homogalactanas sulfatadas e que diferenciavam uma da outra apenas no grau e localização dos grupamentos sulfato. Teste de aPTT mostrou que as frações 2,0 e 3,0 M (Gal 1 e Gal 2, respectivamente) possuem atividade anticoagulante. Esta é a primeira vez que homogalactanas sulfatadas anticoagulantes são extraídas de macroalgas verdes. A massa necessária de Gal 1 (6,3mg) para prolongar o tempo normal de coagulação em duas vezes, via teste de aPTT, foi semelhante a Clexane®, no entanto somente 0,7mg da Gal 2 são necessários para se obter o mesmo efeito. A Gal 2 em altas doses (250mg) induziu a agregação plaquetária. Estes resultados sugerem que as galactanas da alga C. isthmocladum tem potencial aplicação como fármacos anticoagulantes 2014-12-17T14:03:26Z 2006-11-20 2014-12-17T14:03:26Z 2006-07-07 masterThesis FARIAS, Eduardo Henrique Cunha de. Homogalactanas sulfatadas da alga Codium isthmocladum com atividade anticoagulante. 2006. 75 f. Dissertação (Mestrado em Bioquímica; Biologia Molecular) - Universidade Federal do Rio Grande do Norte, Natal, 2006. https://repositorio.ufrn.br/jspui/handle/123456789/12526 por Acesso Aberto application/pdf application/pdf Universidade Federal do Rio Grande do Norte BR UFRN Programa de Pós-Graduação em Bioquímica Bioquímica; Biologia Molecular |