Purificação e caracterização parcial de uma Fucana C de Dictyota mentrualis e estudo do seu efeito antiinflamatório e nociceptivo

In recent years, sulfated polysaccharides from marine algae have emerged as an important class of natural biopolymers with potential application in human and veterinary health care, while taking advantage of the absence of potential risk of contamination by animal viruses. Among these, fucans isolated from the cell walls of marine brown alga have been study due to their anticoagulant, antithrombotic, anti-inflammatory and antiviral activities. These biological effects of fucans have been found to depend on the degree of sulfation and molecular size of the polysaccharide chains. In the present study, we examined structural features of a fucan extracted from brown alga Dictyota menstrualis and its effect on the leukocyte migration to the peritoneum. The sulfated polysaccharides were extracted from the brown seaweed by proteolytic digestion, followed by sequential acetone precipitation producing 5 fractions. Gel lectrophoresis using 0.05 M 1,3-diaminopropane-acetate buffer, pH 9.0, stained with 0.1% toluidine blue, showed the presence of sulfated polysaccharides in all fractions. The chemical analyses demonstrated that all fractions are composed mainly of fucose, xylose, galactose, uronic acid, and sulfate. Electrophoresis in agarose gel in three different buffers demonstrated that the fraction 2.0v have only one population of fucan. This compound was purify by exclusion molecular. It has shown composition of fucose, xilose, sulfate and uronic acid in molar ration of 1.0: 1.7: 1.1: 0.5 respectively. The effect of this heterofucan on the leukocyte migration was observed 6h after zymozan (mg/g) administration into the peritoneum. The heterofucan showed higher antimigratory activity, it decrease the migration of leukocyte in 83.77% to peritoneum. The results suggest that this fucan is a new antimigratory compound with potential pharmacological appicationsEm anos recentes, os polissacarídeos sulfatatos de algas marinhas tem emergido como importante classe natural de biopolimeros que podem ser aplicado na assistencia medica humana e veterinaria, tendoi a vantagem da alsência de potencial risco de contaminação por virus animal. Algumas amostras de fucanas foram isoladas de alga marinha maron tensido estudadas quanto a sua atividader anticoagulante, antitrombótica, antiinflamatória e antiviral. O efeito biológico das fucanas tem sido dependente do grau de sulfatação e do tamanho da molecula do polissacarídeo. Neste estudo foi realizado uma analise da composição da fucana extraída da alga maron Dictyota menstrualis e o efeito na migração de leucócitos no peritonio com sua molulação influencia na modulação da sensação dolorosa periférica. O polissacarídeos sulfatado foi extraído da alga maron por digestão proteolítica sequido por precipitação com acetona produzindo 05 frações. A eletroforese em gel usando tampão 0,05 M 1,3-diaminopropano-acetato (PDA) pH 9,0 revelado com 1,0% de azul de toluidina demonstrou a presença de polissacarídeos em todas as frações. A analise química demonstrou que todas as frações são compostas principalmente de fucose, xilose, galactose, ácido uronico e sulfato. As eletroforeses em três sistemas de tampões demonstraram que a fração 2,0v possui apenas uma população de fucana. Este componente foi purificado por exclusão molecular e demonstrou uma composição de fucose, xilose, sulfato e ácido urônico de 1,0:1,7:1,1:0,5 respectivamente. O efeito desda heterofucana na migração de leucócito observado 06 horas após a aplicação zymozan no peritonio. A heterofucana demonstrou alta atividade antimigratória diminuindo a migração em 100% na concentração de 20,0mg/kg, também apresentamdo um efeito modulado da sensação dolorosa periférica de modo dose dependente. Estes resultados sugerem que este fucana possa ser utilizada como novo antiinflamatório na industria farmaceutic

Evaluation of Anti-Nociceptive and Anti-Inflammatory Activities of a Heterofucan from Dictyota menstrualis

Fucan is a term that defines a family of homo-and hetero-polysaccharides containing sulfated L-fucose in its structure. in this work, a heterofucan (F2.0v) from the seaweed, Dictyota menstrualis, was evaluated as an antinociceptive and anti-inflammatory agent. F2.0v (20.0 mg/kg) inhibits 100% of leukocyte migration into the peritoneal cavity after chemical stimulation. However, F2.0v does not alter the expression of interleukin-1 beta (IL-1 beta) and interleukin-6 (IL-6), as well as tumor necrosis factor alpha (TNF-alpha). F2.0v (20.0 mg/kg) has peripheral antinociceptive activity with potency similar to dipyrone. On the other hand, it had no effect on pain response on the hot plate test. Confocal microscopy analysis and flow cytometry showed that F2.0v binds to the surface of leucocytes, which leads us to suggest that the mechanism of action of anti-inflammatory and antinociceptive F2.0v is related to its ability to inhibit the migration of leukocytes to the site of tissue injury. in summary, the data show that F2.0v compound has great potential as an antinociceptive and anti-inflammatory, and future studies will be performed to further characterize the mechanism of action of F2.0v.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundacao de Apoio a Pesquisa do Estado do Rio Grande do Norte-FAPERNCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Ministerio de Ciencia, Tecnologia e Inovacao-MCTIFed Univ Rio Grande Norte UFRN, Dept Biochem, Lab Biotechnol Nat Polymers BIOPOL, BR-59078970 Natal, RN, BrazilFed Univ Rio Grande Norte UFRN, Grad Program Hlth Sci, BR-59078970 Natal, RN, BrazilFed Univ São Paulo UNIFESP, Dept Biochem, BR-04044020 São Paulo, BrazilFed Univ São Paulo UNIFESP, Dept Biochem, BR-04044020 São Paulo, BrazilWeb of Scienc

Heterofucan from Sargassum filipendula Induces Apoptosis in HeLa Cells

Fucan is a term used to denominate a family of sulfated polysaccharides rich in sulfated l-fucose. Heterofucan SF-1.5v was extracted from the brown seaweed Sargassum filipendula by proteolytic digestion followed by sequential acetone precipitation. This fucan showed antiproliferative activity on Hela cells and induced apoptosis. However, SF-1.5v was not able to activate caspases. Moreover, SF-1.5v induced glycogen synthase kinase (GSK) activation, but this protein is not involved in the heterofucan SF-1.5v induced apoptosis mechanism. In addition, ERK, p38, p53, pAKT and NFκB were not affected by the presence of SF-1.5v. We determined that SF-1.5v induces apoptosis in HeLa mainly by mitochondrial release of apoptosis-inducing factor (AIF) into cytosol. In addition, SF-1.5v decreases the expression of anti-apoptotic protein Bcl-2 and increased expression of apoptogenic protein Bax. These results are significant in that they provide a mechanistic framework for further exploring the use of SF-1.5v as a novel chemotherapeutics against human cervical cancer

Antioxidant and Antiproliferative Activities of Heterofucans from the Seaweed Sargassum filipendula

Fucan is a term used to denominate a type of polysaccharide which contains substantial percentages of l-fucose and sulfate ester groups. We obtained five heterofucans from Sargassum filipendula by proteolytic digestion followed by sequential acetone precipitation. These heterofucans are composed mainly of fucose, glucose, glucuronic acid, galactose and sulfate. These fucans did not show anticoagulant activity in PT and aPTT tests. Their antioxidant activity was evaluated using the follow tests; total antioxidant capacity, scavenging hydroxyl and superoxide radicals, reducing power and ferrous ion [Fe(II)] chelating. All heterofucans displayed considerable activity, especially SF-1.0v which showed the most significant antioxidant potential with 90.7 ascorbic acid equivalents in a total antioxidant capacity test and similar activity when compared with vitamin C in a reducing power assay. The fucan antiproliferative activity was performed with HeLa, PC3 and HepG2 cells using MTT test. In all tested conditions the heterofucans exhibited a dose-dependent effect. The strongest inhibition was observed in HeLa cells, where SF-1.0 and SF-1.5 exhibited considerable activity with an IC50 value of 15.69 and 13.83 μM, respectively. These results clearly indicate the beneficial effect of S. filipendula polysaccharides as antiproliferative and antioxidant. Further purification steps and additional studies on structural features as well as in vivo experiments are needed to test the viability of their use as therapeutic agents

Anticoagulant, Antioxidant and Antitumor Activities of Heterofucans from the Seaweed Dictyopteris delicatula

In the present study, six families of sulfated polysaccharides were obtained from seaweed Dictyopteris delicatula by proteolytic digestion, followed by acetone fractionation and molecular sieving on Sephadex G-100. Chemical analyses demonstrated that all polysaccharides contain heterofucans composed mainly of fucose, xylose, glucose, galactose, uronic acid, and sulfate. The fucans F0.5v and F0.7v at 1.0 mg/mL showed high ferric chelating activity (~45%), whereas fucans F1.3v (0.5 mg/mL) showed considerable reducing power, about 53.2% of the activity of vitamin C. The fucan F1.5v presented the most prominent anticoagulant activity. The best antiproliferative activity was found with fucans F1.3v and F0.7v. However, F1.3v activity was much higher than F0.7v inhibiting almost 100% of HeLa cell proliferation. These fucans have been selected for further studies on structural characterization as well as in vivo experiments, which are already in progress