The results indicate that the analgesic and anti-inflammatory effects of Cc-SP2 could be of biomedical applicability as a new, natural tool in pain and acute inflammatory conditions.
RESUMO Este estudo teve como objetivo isolar, fracionar e avaliar o potencial anticoagulante de iota-carragenanas (iCARs) da rodofícea Solieria filiformis, quando obtidas por dois métodos de extração (M I e M II). As i-CARs foram isoladas ABSTRACT This study aimed to isolate, fractionate and evaluate the anticoagulant potential of iota-carrageenans (i-CARs) from Solieria filiformis when two extraction methods (M I and M II) were used. i-CARs were isolated with papain in 0.1M sodium acetate (pH 5.0) containing 5mM cystein and 5mM EDTA (M I) or water (80°C) (M II), and then their chemical composition of total carbohydrates, free sulfate (FS) and contaminant proteins were determined. i-CARs were submitted to anionexchange chromatography (DEAE-cellulose) using a sodium chloride gradient,being evaluated the activated partial thromboplastin time (APTT) and prothrombin time of obtained fractions and compared to heparin (
ABSTRACT. Biochemical analyses are important tools for discovering new bioactive compounds for medical clinic. This study aimed at isolating iota-carrageenans (ι-CARs) from Solieria filiformis (Rhodophyta) by enzymatic extraction (EE), refined hot-water extraction (RHWE) and hot-water extraction (HWE), and test (EE, s.c.) their anti-inflammatory effects in the peritonitis model using the Lambda-carrageenan (700 μg cavity -1 , i.p.) as an inflammatory stimuli in rats. The activated partial thromboplastin time (APTT) was also evaluated in ι-CARs fractions, obtained by ion-exchange chromatography (DEAE-cellulose), using rabbit plasma and compared to heparin ). The results showed that the ι-CARs (EE) (3 or 9 mg kg -1 ) containing 89.92% total sugars, 29.02% sulfate and absence of contaminant proteins inhibited (p < 0.05) the cellular infiltrate in the peritoneal cavity of the animals, but with 27 mg kg Iota-carragenanas da rodofícea Solieria filiformis e seus efeitos na inflamação e coagulação RESUMO. Análises bioquímicas são ferramentas importantes para a descoberta de novos compostos bioativos para clínica médica. Neste estudo, iota-carragenanas (ι-CARs) da rodofícea Solieria filiformis foram isoladas (extração enzimática (EE), extração aquosa a quente refinada (EAQR) ou extração aquosa a quente (EAQ)) e testadas (EE; s.c.) no modelo de peritonite, usando-se a Lambda-carragenana (700 μg cavidade -1 ; i.p.) como um estímulo flogístico, para averiguar seus efeitos anti-inflamatórios em ratos. O tempo de tromboplastina parcial ativada (TTPA) também foi avaliado em frações de ι-CARs, obtidas por cromatografia de troca iônica (DEAE-celulose), usando plasma de coelho e comparadas à heparina ). Verificaram-se que ι-CARs (EE) (3 ou 9 mg kg -1 ) contendo 89,92% de açúcares totais, 29,02% de sulfato e destituídas de contaminação proteica, inibiram (p < 0,05) o infiltrado celular na cavidade peritoneal dos animais, mas com 27 mg kg -1 não apresentaram efeito anti-inflamatório (p > 0,05). Os perfis cromatográficos mostraram-se semelhantes entre os métodos de extração, porém revelando, por eletroforese, frações com diferenças em termos de densidade de carga. As frações praticamente não alteraram o TTPA (1,16; 1,73 e 1,59 UI mg -1 para EE, EAQR e EAQ, respectivamente). Investigações são sugeridas para melhor compreender as ações das ι-CARs (EE) da S. filiformis na resposta inflamatória.Palavras-chave: alga vermelha, galactanas sulfatadas, métodos de extração, peritonite, teste do TTPA.
RESUMO.O cultivo da rodofícea Solieria filiformis vem sendo desenvolvido na praia de Flecheiras, Estado do Ceará, como perspectiva de utilização sustentável do recurso para biotecnologia, mas o crescimento de epífitas sobre a espécie compromete seu bom crescimento. Desta forma, avaliou-se a eficiência de duas metodologias (M I e M II) no isolamento da Kappa-carragenana (κ-CAR) da alga marinha vermelha Hypnea musciformis. A κ-CAR foi obtida mediante digestão proteolítica (M I) ou água quente (80°C) (M II). Os extratos foram analisados quanto à sua composição química, seguido por cromatografia de troca iônica em coluna de DEAE-celulose usando um gradiente de NaCl. A atividade anticoagulante foi avaliada pelo teste do tempo de tromboplastina parcial ativada (TTPA), usando plasma humano normal e heparina (HEP) padrão (193 UI mg ABSTRACT. Carragennan from Hypnea musciformis epiphyte obtained fromSolieria filiformis experimental culture at Flecheiras, Ceará State, Brazil. Solieria filiformis (Rhodophyceae) culture is being developed at Flecheiras Beach, state of Ceará, Brazil, for its future sustainable use in biotechnology. However, epiphyte growth on the species jeopardizes suitable development. The efficiency of two methods (M I and M II) in the isolation of kappa-carrageenan (κ-CAR) from red marine alga Hypnea musciformis has been evaluated. κ-CAR was obtained by proteolytic digestion (M I) or by hot water (80°C) (M II) and extracts were analyzed with regard to their chemical composition and then by ion-exchange chromatography on DEAE-cellulose column with NaCl gradient. Anticoagulant activity was evaluated by the activated partial thromboplastine time (APTT) test with common human plasma and standard heparin (HEP) ). Yield difference reached 4.28 ± 0.26%% (p < 0.05), and the lowest sulfate content was obtained with M I (17.00%) when compared to that with M II (26.30%). Further, different chromatographic profiles were reported between the two methods. Whereas M I was efficacious in obtaining κ-CAR anticoagulants (2.67 IU mg -1 ), alternative M II was efficient for the food industry. Results suggest that κ-CAR extraction method from H. musciformis depends on the industry's usage criterion.
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