Aspectos estruturais, farmacológicos e biológicos de fucanas da alga marrom sargassum vulgare

Detalhes bibliográficos
Autor(a) principal: Dore, Celina Maria Pinto Guerra
Data de Publicação: 2012
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFRN
Texto Completo: https://repositorio.ufrn.br/jspui/handle/123456789/12570
Resumo: The present study examines the chemical composition and their effects on free radicals, inflammation, angiogenesis, coagulation, VEGF effects and cellular proliferation of a polysaccharides from alga Sargassum vulgare. The sulfated polysaccharide was extracted from brown seaweed by proteolysis with enzymes maxataze. The presence of proteins and sugars were observed in crude polysaccharides. Fractionation of this crude extract was made with growing concentration of acetone (0.3-1.5 v) and produced four groups of polysaccharides. Anionic polysaccharides from brown seaweed Sargassum vulgare, SV1and PSV1 were fractionated (SV1) and purified (PSV1), and displayed with high total sugars and sulfate content and very low level of protein. This fucan SV1 contains low levels of protein and high carbohydrate and sulfate content. This polysaccharides prolonged activated partial thromboplastin time (aPTT) at 50 μg (>240 s). SV1 was found to have no effect on prothrombin time (PT), corresponding to the extrinsic pathway of coagulation. SV1 exhibits high antithrombotic action in vivo, with a concentration ten times higher than heparin. Polysaccharides from S. vulgare promoted direct inhibition enzymatic activity of thrombin and stimulated enzymatic activity of FXa. SV1 showed optimal inhibitory activity of thrombin (50.2±0.28%) at a concentration of 25 μg/mL. Its antioxidant action on scavenging radicals by DPPH was (22%), indicating the polymer has no cytotoxic action (hemolytic) on ABO and Rh blood types in different erythrocyte groups and displays strong anti-inflammatory action on all concentrations tested in the carrageenan-induced paw edema model, demonstrated by reduced edema and cellular infiltration. Angiogenesis is a dynamic process of proliferation and differentiation. It requires endothelial proliferation, migration, and tube formation. In this context, endothelial cells are a preferred target for several studies and therapies. The antiangiogenic efficacy of polysaccharides was examined in vivo in the chick chorioallantoic membrane (CAM) model by using fertilized eggs. Decreases in the density of the capillaries were assessed and scored. The results showed that SV1 and PSV1 have an inhibitory effect on angiogenesis. These results were also confirmed by inhibition tubulogenesis in rabbit aorta endothelial cell (RAEC) in matrigel. These compounds were assessed in Apoptosis assay (Annexin V - FITC / PI) and cell viability by MTT assay of RAEC. These polysaccharides do not affect the viability and do not have apoptotic or necrotic action. RAEC cell when incubated with SV1 and PSV1showed inhibition of VEGF secretion, observed when compounds were incubated at 25, 50 and 100 μg/μL. The VEGF secretion with the RAEC cell line for 24 h, was more effective for PSV1 at 50 μg/μL(71.4%) than SV1 100 μg/μL (75.9%). SV1 and PSV1 had an antiproliferative action (47%) against tumor cell line HeLa. Our results indicate that these sulfated polysaccharides have antiangiogenic and antitumoral actions
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spelling Dore, Celina Maria Pinto Guerrahttp://lattes.cnpq.br/2279068301048550http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783254U6&dataRevisao=nullCruz, Ana Katarina Menezes dahttp://lattes.cnpq.br/9810605697247961Farias, Eduardo Henrique Cunha dehttp://lattes.cnpq.br/0933304924768138Filgueira, Luciana Guimarães Alveshttp://lattes.cnpq.br/9951316929526841Carvalho, Maria Goretti Freire dehttp://lattes.cnpq.br/8934375314306198Leite, Edda Lisboa2014-12-17T14:03:35Z2013-04-152014-12-17T14:03:35Z2012-10-15DORE, Celina Maria Pinto Guerra. Aspectos estruturais, farmacológicos e biológicos de fucanas da alga marrom sargassum vulgare. 2012. 112 f. Tese (Doutorado em Bioquímica; Biologia Molecular) - Universidade Federal do Rio Grande do Norte, Natal, 2012.https://repositorio.ufrn.br/jspui/handle/123456789/12570The present study examines the chemical composition and their effects on free radicals, inflammation, angiogenesis, coagulation, VEGF effects and cellular proliferation of a polysaccharides from alga Sargassum vulgare. The sulfated polysaccharide was extracted from brown seaweed by proteolysis with enzymes maxataze. The presence of proteins and sugars were observed in crude polysaccharides. Fractionation of this crude extract was made with growing concentration of acetone (0.3-1.5 v) and produced four groups of polysaccharides. Anionic polysaccharides from brown seaweed Sargassum vulgare, SV1and PSV1 were fractionated (SV1) and purified (PSV1), and displayed with high total sugars and sulfate content and very low level of protein. This fucan SV1 contains low levels of protein and high carbohydrate and sulfate content. This polysaccharides prolonged activated partial thromboplastin time (aPTT) at 50 μg (>240 s). SV1 was found to have no effect on prothrombin time (PT), corresponding to the extrinsic pathway of coagulation. SV1 exhibits high antithrombotic action in vivo, with a concentration ten times higher than heparin. Polysaccharides from S. vulgare promoted direct inhibition enzymatic activity of thrombin and stimulated enzymatic activity of FXa. SV1 showed optimal inhibitory activity of thrombin (50.2±0.28%) at a concentration of 25 μg/mL. Its antioxidant action on scavenging radicals by DPPH was (22%), indicating the polymer has no cytotoxic action (hemolytic) on ABO and Rh blood types in different erythrocyte groups and displays strong anti-inflammatory action on all concentrations tested in the carrageenan-induced paw edema model, demonstrated by reduced edema and cellular infiltration. Angiogenesis is a dynamic process of proliferation and differentiation. It requires endothelial proliferation, migration, and tube formation. In this context, endothelial cells are a preferred target for several studies and therapies. The antiangiogenic efficacy of polysaccharides was examined in vivo in the chick chorioallantoic membrane (CAM) model by using fertilized eggs. Decreases in the density of the capillaries were assessed and scored. The results showed that SV1 and PSV1 have an inhibitory effect on angiogenesis. These results were also confirmed by inhibition tubulogenesis in rabbit aorta endothelial cell (RAEC) in matrigel. These compounds were assessed in Apoptosis assay (Annexin V - FITC / PI) and cell viability by MTT assay of RAEC. These polysaccharides do not affect the viability and do not have apoptotic or necrotic action. RAEC cell when incubated with SV1 and PSV1showed inhibition of VEGF secretion, observed when compounds were incubated at 25, 50 and 100 μg/μL. The VEGF secretion with the RAEC cell line for 24 h, was more effective for PSV1 at 50 μg/μL(71.4%) than SV1 100 μg/μL (75.9%). SV1 and PSV1 had an antiproliferative action (47%) against tumor cell line HeLa. Our results indicate that these sulfated polysaccharides have antiangiogenic and antitumoral actionsO presente estudo analisa a composição química e seus efeitos sobre os radicais livres, inflamação, angiogênese, coagulação, VEGF e proliferação celular dos polissacarídeos de uma alga Sargassum vulgare. O polissacárido sulfatado foi extraído a partir de algas marrons por proteólise com a enzima maxataze. A presença de proteínas e açúcares foram observados no cru de polissacarideos. Fracionamento do o extrato bruto foi feito com concentrações crescente de acetona (0,3-1,5 v), produzindo quatro grupos de polissacarideos. Estes compostos aniônicos da alga S. vulgare, foram fracionados (SV1) e purificados (PSV1) exibindo com alta açúcares totais e sulfatecontent e nível muito baixo de proteínas.A fucana SV1 contém baixos níveis de proteína e de hidratos de carbono e alto teor de sulfato. Este polissacarídeos prolongou o tempo de tromboplastina parcial activada (aPTT) a 50 ug (>240 s). não foi observado qualquer efeito de SV1 sobre o tempo de protrombina (PT), que corresponde a via extrínseca da coagulação. SV1 exibiu alta ação antitrombótica in vivo, com uma concentração 10 vezes maior do que a heparina. SV1 promoveu a actividade de inibição enzimática direta da trombina e estimulou a atividade enzimática do FXa. Mostrou também, atividade inibidora optima de trombina (50,2 ± 0,28%) a uma concentração de 25 ug / mL. A sua acção anti-oxidante de radicais scavenging por DPPH foi de (22%), indicando que o polímero não tem qualquer ação citotóxica (hemolítica) em tipos de sangue ABO e Rh, em diferentes grupos de eritrócitos e exibindo alta ação anti-inflamatória em edema de pata de ratos Wistar em todas as concentrações testadas induzida por carragenina. Tal processo foi demonstrado por edema e infiltração celular. A angiogenese é um processo dinâmico de proliferação e diferenciação. Ele requer proliferação endotelial, migração, e a formação do tubo. Neste contexto, as células endoteliais são um alvo preferido para muitos estudos e terapias. A eficácia antiangiogenico de polissacarídeos foi examinada in vivo na membrana corioalantóica pinto (CAM) usando-se ovos fertilizados. Diminuições na densidade dos capilares foram avaliados e pontuados. Os resultados mostraram que SV1 e PSV1 tem um efeito inibidor da angiogenese. Estes resultados foram também confirmados por tubulogenesis inibição na célula endotelial da aorta de coelho (RAEC) em matrigel. Células RAEC quando foram incubadas com SV1and PSV1 demonstraram inibição da secreção de VEGF, a 25, 50 e 100 ug/mL. A secreção de VEGF com a linha de células RAEC durante 24 h, foi mais eficaz para PSV1 a 50 ug / mL (71,4%) do que SV1 100 ug / mL (75,9%). SV1 e PSV1 posuiram uma acção antiproliferativa (47%) contra as células tumorais tipo HeLa. Estes compostos foram avaliados também, no ensaio de apoptose (anexina V - FITC / PI) e a viabilidade celular pelo ensaio de MTT de RAEC. Estes polissacarídeos não afetaram a viabilidade e não tiveram ação apoptótica ou necrótica. Nossos resultados indicam que estes polissacarídeos sulfatados têm ações antiangiogênica e antitumoral e constituem um importante alvo biológico e farmacológicoConselho Nacional de Desenvolvimento Científico e Tecnológicoapplication/pdfporUniversidade Federal do Rio Grande do NortePrograma de Pós-Graduação em BioquímicaUFRNBRBioquímica; Biologia MolecularAlga marrom. Fucana. Hemostasia. AngiogêneseBrown seaweed. Fucan. Hemostasis. 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dc.title.por.fl_str_mv Aspectos estruturais, farmacológicos e biológicos de fucanas da alga marrom sargassum vulgare
title Aspectos estruturais, farmacológicos e biológicos de fucanas da alga marrom sargassum vulgare
spellingShingle Aspectos estruturais, farmacológicos e biológicos de fucanas da alga marrom sargassum vulgare
Dore, Celina Maria Pinto Guerra
Alga marrom. Fucana. Hemostasia. Angiogênese
Brown seaweed. Fucan. Hemostasis. Angiogenesis
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
title_short Aspectos estruturais, farmacológicos e biológicos de fucanas da alga marrom sargassum vulgare
title_full Aspectos estruturais, farmacológicos e biológicos de fucanas da alga marrom sargassum vulgare
title_fullStr Aspectos estruturais, farmacológicos e biológicos de fucanas da alga marrom sargassum vulgare
title_full_unstemmed Aspectos estruturais, farmacológicos e biológicos de fucanas da alga marrom sargassum vulgare
title_sort Aspectos estruturais, farmacológicos e biológicos de fucanas da alga marrom sargassum vulgare
author Dore, Celina Maria Pinto Guerra
author_facet Dore, Celina Maria Pinto Guerra
author_role author
dc.contributor.authorID.por.fl_str_mv
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/2279068301048550
dc.contributor.advisorID.por.fl_str_mv
dc.contributor.advisorLattes.por.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783254U6&dataRevisao=null
dc.contributor.referees1.pt_BR.fl_str_mv Cruz, Ana Katarina Menezes da
dc.contributor.referees1ID.por.fl_str_mv
dc.contributor.referees1Lattes.por.fl_str_mv http://lattes.cnpq.br/9810605697247961
dc.contributor.referees2.pt_BR.fl_str_mv Farias, Eduardo Henrique Cunha de
dc.contributor.referees2ID.por.fl_str_mv
dc.contributor.referees2Lattes.por.fl_str_mv http://lattes.cnpq.br/0933304924768138
dc.contributor.referees3.pt_BR.fl_str_mv Filgueira, Luciana Guimarães Alves
dc.contributor.referees3ID.por.fl_str_mv
dc.contributor.referees3Lattes.por.fl_str_mv http://lattes.cnpq.br/9951316929526841
dc.contributor.referees4.pt_BR.fl_str_mv Carvalho, Maria Goretti Freire de
dc.contributor.referees4ID.por.fl_str_mv
dc.contributor.referees4Lattes.por.fl_str_mv http://lattes.cnpq.br/8934375314306198
dc.contributor.author.fl_str_mv Dore, Celina Maria Pinto Guerra
dc.contributor.advisor1.fl_str_mv Leite, Edda Lisboa
contributor_str_mv Leite, Edda Lisboa
dc.subject.por.fl_str_mv Alga marrom. Fucana. Hemostasia. Angiogênese
topic Alga marrom. Fucana. Hemostasia. Angiogênese
Brown seaweed. Fucan. Hemostasis. Angiogenesis
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
dc.subject.eng.fl_str_mv Brown seaweed. Fucan. Hemostasis. Angiogenesis
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
description The present study examines the chemical composition and their effects on free radicals, inflammation, angiogenesis, coagulation, VEGF effects and cellular proliferation of a polysaccharides from alga Sargassum vulgare. The sulfated polysaccharide was extracted from brown seaweed by proteolysis with enzymes maxataze. The presence of proteins and sugars were observed in crude polysaccharides. Fractionation of this crude extract was made with growing concentration of acetone (0.3-1.5 v) and produced four groups of polysaccharides. Anionic polysaccharides from brown seaweed Sargassum vulgare, SV1and PSV1 were fractionated (SV1) and purified (PSV1), and displayed with high total sugars and sulfate content and very low level of protein. This fucan SV1 contains low levels of protein and high carbohydrate and sulfate content. This polysaccharides prolonged activated partial thromboplastin time (aPTT) at 50 μg (>240 s). SV1 was found to have no effect on prothrombin time (PT), corresponding to the extrinsic pathway of coagulation. SV1 exhibits high antithrombotic action in vivo, with a concentration ten times higher than heparin. Polysaccharides from S. vulgare promoted direct inhibition enzymatic activity of thrombin and stimulated enzymatic activity of FXa. SV1 showed optimal inhibitory activity of thrombin (50.2±0.28%) at a concentration of 25 μg/mL. Its antioxidant action on scavenging radicals by DPPH was (22%), indicating the polymer has no cytotoxic action (hemolytic) on ABO and Rh blood types in different erythrocyte groups and displays strong anti-inflammatory action on all concentrations tested in the carrageenan-induced paw edema model, demonstrated by reduced edema and cellular infiltration. Angiogenesis is a dynamic process of proliferation and differentiation. It requires endothelial proliferation, migration, and tube formation. In this context, endothelial cells are a preferred target for several studies and therapies. The antiangiogenic efficacy of polysaccharides was examined in vivo in the chick chorioallantoic membrane (CAM) model by using fertilized eggs. Decreases in the density of the capillaries were assessed and scored. The results showed that SV1 and PSV1 have an inhibitory effect on angiogenesis. These results were also confirmed by inhibition tubulogenesis in rabbit aorta endothelial cell (RAEC) in matrigel. These compounds were assessed in Apoptosis assay (Annexin V - FITC / PI) and cell viability by MTT assay of RAEC. These polysaccharides do not affect the viability and do not have apoptotic or necrotic action. RAEC cell when incubated with SV1 and PSV1showed inhibition of VEGF secretion, observed when compounds were incubated at 25, 50 and 100 μg/μL. The VEGF secretion with the RAEC cell line for 24 h, was more effective for PSV1 at 50 μg/μL(71.4%) than SV1 100 μg/μL (75.9%). SV1 and PSV1 had an antiproliferative action (47%) against tumor cell line HeLa. Our results indicate that these sulfated polysaccharides have antiangiogenic and antitumoral actions
publishDate 2012
dc.date.issued.fl_str_mv 2012-10-15
dc.date.available.fl_str_mv 2013-04-15
2014-12-17T14:03:35Z
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dc.identifier.citation.fl_str_mv DORE, Celina Maria Pinto Guerra. Aspectos estruturais, farmacológicos e biológicos de fucanas da alga marrom sargassum vulgare. 2012. 112 f. Tese (Doutorado em Bioquímica; Biologia Molecular) - Universidade Federal do Rio Grande do Norte, Natal, 2012.
dc.identifier.uri.fl_str_mv https://repositorio.ufrn.br/jspui/handle/123456789/12570
identifier_str_mv DORE, Celina Maria Pinto Guerra. Aspectos estruturais, farmacológicos e biológicos de fucanas da alga marrom sargassum vulgare. 2012. 112 f. Tese (Doutorado em Bioquímica; Biologia Molecular) - Universidade Federal do Rio Grande do Norte, Natal, 2012.
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