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Juliana Silva Nascimento De Novais Caracterização Da Matéria Prima Universidade Federal do Rio de Janeiro Programa de Pós-graduação em Produtos Bioativos e Biociências Juliana Silva Nascimento De Novais Caracterização da matéria prima vegetal e atividade biológica das folhas de Senna alata (L.) Roxb. Em acessos na caatinga (BA) e restinga (RJ), Brasil MACAÉ-RJ, 2017 Universidade Federal do Rio de Janeiro Programa de Pós-graduação em Produtos Bioativos e Biociências Juliana Silva Nascimento De Novais Caracterização da matéria prima vegetal e atividade biológica das folhas de Senna alata (L.) Roxb. Em acessos na caatinga (BA) e restinga (RJ), Brasil Dissertação a ser apresentada pela aluna Juliana Silva Nascimento de Novais ao Programa de Pós Graduação em Produtos Bioativos e Biociencias, da Universidade Federal do Rio de Janeiro, Campus UFRJ Macaé, como requisito parcial para a obtenção do título de Mestre em Ciências. Orientador: Prof Dr Gilberto Dolejal Zanetti Co-orientação: Prof. Drª Elaine dos Anjos da Cruz da Rocha Macaé, RJ 2017 CARACTERIZAÇÃO DA MATÉRIA PRIMA VEGETAL E ATIVIDADE BIOLÓGICA DAS FOLHAS DE SENNA ALATA (L.) ROXB. EM ACESSOS NA CAATINGA (BA) E RESTINGA (RJ), BRASIL Juliana Silva Nascimento de Novais Dissertação apresentada ao Programa de Pós Graduação em Produtos Bioativos e Biociencias da Universidade Federal do Rio de Janeiro, como parte das exigências para a obtenção do título de Mestre em Ciências. Aprovada em 30 de Outubro de 2017. _______________________________________________ Prof Dr Arídio Mattos Junior _______________________________________________ Prof Dr Edison Luis Santana de Carvalho _______________________________________________ Prof Dra Michelle Frazão Muzitano ________________________________________________ Orientador: Prof Dr Gilberto Dolejal Zanetti _______________________________________________ Coorientadora: Profa Dra Elaine dos Anjos da Cruz da Rocha Macaé, RJ 2017 AGRADECIMENTOS Agradeço a Deus por ter me presenteado com o dom da vida. Gostaria de agradecer a diversas pessoas pela ajuda, apoio, contribuição, atenção e paciência, sem os quais tornaria a realização desse trabalho imensamente árduo. Estas são: Meu esposo, por apoiar, participar e está sempre disposto a me ajudar em uma participação direta na concretização dos meus sonhos. Pela paciência durante incontáveis horas em que tive que está no laboratório ou escrevendo durante madrugadas. Pelo amor que ameniza a dor e torna tudo possível. À minha mãe e minha irmã pelo amor incondicional, pelas palavras doces e duras de amor e de encorajamento, por sempre se fazerem tão presentes mesmo a distância sendo um abismo. Sou grata ao meu orientador Gilberto Zanetti por nunca escolher o caminho mais fácil ou conveniente, mas por sempre escolher o caminho certo. Por ter acreditado em mim quando eu própria não podia. Pelo carinho e amizade dispensados a mim. Por cumprir de maneira graciosa e exemplar o sentido da palavra “mentor” na sua definição mais completa. Pela sua brilhante capacidade científica. Pela sua incansável dedicação no desenvolvimento da carreira de cada um dos seus alunos. Pelas suas aulas que iam além do assunto proposto, de forma didática e interessante. Pelo exemplo de inspiração. Agradeço também a minha coorientadora Elaine da Rocha, por generosamente abrir as portas do seu laboratório e compartilhar seu conhecimento e recurso no desenvolvimento desse trabalho. Por ter me abraçado e por não ter medido esforços em me ajudar e apoiar. Por não olhar simplesmente naquilo que se investiga, mas se preocupar de forma generosa com quem está investigando. Pelo exemplo de paciência, sabedoria e liderança. Aos técnicos e amigos do Instituto de ciência e tecnologia de Macaé, em especial, Jobert e Jéssyca, que foram sempre presentes durante esses anos e me ajudaram no desenvolvimento desse trabalho. Aos docentes, Denise, Edison e Michelle, por me ensinarem que além de cérebros brilhantes haviam corações generosos e dispostos a ajudar. Por toda auxílio, conselhos e apoio no desenvolvimento desse estudo. Ao professor Arídio que aceitou contribuir com esse trabalho. Agradeço universidade de Taubaté em especial a colaboração da professora Josseara Beralto, que mesmo sem me conhecer me apoiou no que foi preciso para concluir os experimentos. A equipe da secretaria de coordenação do PRODGRADBIO, principalmente, a Andreia e o Leonardo pela atenção, paciência e carinho. Enfim a todas as pessoas que contribuíram direta e indiretamente na realização desse trabalho e que porventura tenham sido omitidas, meus sinceros agradecimentos. RESUMO NOVAIS, Juliana Silva Nascimento. Caracterização da Matéria prima vegetal e atividade biológica das folhas de Senna alata (L.) Roxb. em acessos na caatinga (BA) e restinga (RJ), Brasil. Macaé, RJ, 2017. Dissertação (Mestrado em ciências) – Universidade Federal do Rio de Janeiro, Macaé, 2017. Senna alata (L.) Roxb. é uma leguminosa arbustiva da família Fabaceae introduzida na caatinga e na restinga do Brasil. Essa espécie possui utilização tradicional para fins medicinais o que motivou este estudo que visa caracterizar as folhas e identificar, comparativamente, as atividades biológicas de Senna alata (L.) Roxb. em acessos na caatinga (BA) e restinga (RJ). Após a coleta, o material foi desidratado e levado a pó. Obtiveram-se extratos etanólicos a 40 % por maceração e, este extrato bruto seco, foi resuspenso em diferentes solventes de acordo com o ensaio a ser realizado. Optou-se pelo extrato etanólico a 40 % uma vez que os acessos apresentaram maior teor extrativo nesta concentração de solventes. As folhas nos dois acessos apresentaram morfo-anatomia semelhante pelas técnicas farmacopeias, destacando-se como marcador os tricomas tectores e a presença de estômatos nas duas faces da epiderme. Também se obteve resultados de pureza satisfatórios aos padrões farmacopeicos. Detectou-se flavonoides, saponinas hemolíticas, antraquinonas e mucilagem para os dois acessos e taninos apenas no acesso da restinga. O teor total de polifenois e de flavonoides foi semelhante nos dois acessos. O perfil cromatográfico foi obtido em cromatoplacas de sílica gel com sistema eluente constituído por hexano: acetato de etila (1:1) e vanilina sulfúrica mantida a 100º C por 5 minutos como agente cromogênico, revelando 2 bandas semelhantes para os acessos. A determinação da atividade antibacteriana por bioautografia demonstrou que os dois acessos inibiram o crescimento de Staphylococcus aureus e Escherichia coli. A atividade alelopática frente a diásporos de Lactuca sativa demonstrou potencial alelopático em extratos nas concentrações a partir de 0,5 %, sujerindo atividade inseticida e herbicida. O extrato dos acessos não demonstrou citotoxidade nas concentrações 50 e 100 μg/mL pelo método de {brometo de [3-(4,5-dimetiltiazol-2-il)-2,5-difenil tetrazólio]}, nas concentrações testadas. A atividade anti-inflamatória avaliada pela dosagem de óxido nítrico e fator de necrose tumoral alfa por células intraperitoneais de camundongo Swiss, induzido por lipopolissacarídeo e o polissacarídeo Zymosan. Observou-se um potencial efeito anti- inflamatório sobre macrófagos peritoneais quando estimulados com lipopolissacarídeo e tratados com os 2 acessos na concentração 100 µg/mL, e em macrófagos estimulados por Zymosan, após tratamento a partir de 50 µg/mL do acesso caatinga e 100 µg/mL do acesso restinga. Os extratos apresentam baixa toxidez aguda via intraperitoneal em camundongos Swiss, com concentração letal de 40 e 60 g/kg nos acessos da caatinga e restinga, respectivamente e os níveis de lactato desidrogenase mantiveram-se normais. Os resultados obtidos foram semelhantes para os acessos de Senna alata na caatinga e restinga, sendo que os extratos estudados revelaram baixa toxidez e resultados promissores nas atividades biológicas testadas, conferindo para as folhas de à Senna alata potencialidade real no desenvolvimento de um fitoterápico. Palavras chaves: Fabaceae; Senna alata; Farmacobotânica; Plantas medicinais; Atividades biológicas; Anti-inflamatório; Toxidez; Citotoxidade. ABSTRACT NOVAIS, Juliana Silva Nascimento. Characterization of vegetable raw materials and biological activity of the leaves of Senna alata (L.) Roxb. in access in the caatinga (BA) and restinga (RJ), Brazil. Macaé, RJ, 2017. Dissertation (master of science)-Federal University of Rio de Janeiro, Macaé, 2017. Senna alata (L.) Roxb. is a shrubby legume of the Fabaceae family introduced in the caatinga and the restinga areas of Brazil. This species is traditionally used for medicinal purposes. Therefore this study aims to compare the characterization and identification of leaves, the biological activities of Senna alata (L.) Roxb. in access from the caatinga (Bahia) and restinga (Rio de Janeiro). After collection, the material was dehydrated and taken to dust. Ethanolic extracts were obtained at concentration of 40% by maceration. This crude extract, was ressuspended in various solvents according to the test being performed. We opted for the 40% ethanolic extract once the hits presentedthe most extractive content in this concentration of solvents. The leaves showed morpho-anatomy by using Pharmacopoeias techniques, including tectores and estomatic trichomes types. It was also observed satisfactory results of purity according to Pharmacopeia standards. It were detected flavonoids, haemolytic saponins, anthraquinones and mucilage for both access and tannins only in resting access. The total content of polyphenols and flavonoids was similar both accesses. The chromatographic profile was retrieved from cromatoplates of silica gel with
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