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Universidade Federal De Pernambuco Centro De Ciências Da Saúde Departamento De Ciências Farmacêuticas Programa De Pós-Graduação Em Ciências Farmacêuticas

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Universidade Federal De Pernambuco Centro De Ciências Da Saúde Departamento De Ciências Farmacêuticas Programa De Pós-Graduação Em Ciências Farmacêuticas UNIVERSIDADE FEDERAL DE PERNAMBUCO CENTRO DE CIÊNCIAS DA SAÚDE DEPARTAMENTO DE CIÊNCIAS FARMACÊUTICAS PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS FARMACÊUTICAS RAÍRA JUSTINO OLIVEIRA COSTA Perfil fitoquímico do extrato etanólico das folhas de Jatropha mollissima (Pohl) Baill. e avaliação da sua atividade toxicológica, antitumoral e antibacteriana Recife, 2018 RAÍRA JUSTINO OLIVEIRA COSTA Perfil fitoquímico do extrato etanólico das folhas de Jatropha mollissima (Pohl) Baill. e avaliação da sua atividade toxicológica, antitumoral e antibacteriana Dissertação apresentada ao Programa de Pós-graduação em Ciências Farmacêuticas, da Universidade Federal de Pernambuco, como parte dos requisitos necessários à obtenção do título de Mestre em Ciências Farmacêuticas. Área de concentração: Produtos Naturais e seus compostos bioativos. Orientadora: Prof.ª Dr. ª Ivone Antônia de Souza Coorientadora: Prof.ª Dr.ª Maria Aparecida Barreto Lopes Seabra Recife, 2018 Catalogação na fonte: bibliotecário: Aécio Oberdam, CRB4:1895 C837p Costa, Raíra Justino Oliveira. Perfil fitoquímico do extrato etanólico das folhas de Jatropha mollissima (Pohl) Baill. e avaliação da sua atividade toxicológica, antitumoral e antibacteriana / Raíra Justino Oliveira Costa. – Recife: o autor, 2018. 81 f.; il; 30 cm. Orientadora: Ivone Antônia de Souza. Dissertação (mestrado) – Universidade Federal de Pernambuco. Centro de Ciências da Saúde. Programa de pós-graduação em Ciências Farmacêuticas. Inclui referências, figuras, tabelas e gráficos. 1. Extratos Vegetais. 2. Histologia. 3. Resistência Microbiana a Medicamentos. 4. Sinergia de Substâncias Bioativas. 5. Testes de Drogas Anticâncer. I. Souza, Ivone Antônia. (orientadora). II. Título. 615.1 CDD (23.ed.) UFPE (CCS 2018 - 104) RAÍRA JUSTINO OLIVEIRA COSTA Perfil fitoquímico do extrato etanólico das folhas de Jatropha mollissima (Pohl) Baill. e avaliação da sua atividade toxicológica, antitumoral e antibacteriana Dissertação apresentada ao Programa de Pós-graduação em Ciências Farmacêuticas, da Universidade Federal de Pernambuco, como parte dos requisitos necessários à obtenção do título de Mestre em Ciências Farmacêuticas. Dissertação aprovada em: 27 / 02 / 2018. BANCA EXAMINADORA _____________________________________________________________ Prof.ª Dr.ª Ivone Antônia de Souza (Orientadora) Universidade Federal de Pernambuco _____________________________________________________________ Prof.ª Dr.ª Rosali Maria Ferreira da Silva (Examinador Interno) Universidade Federal de Pernambuco _____________________________________________________________ Prof.ª Dr.ª Jane Sheila Higino (Examinador Externo) Universidade Federal de Pernambuco AGRADECIMENTOS A Deus inicialmente, por sempre se apresentar como um apoio nas horas que só a sua presença me confortaria. Aos meus pais, Francisca e Lairton, por todos esforços realizados para que eu alcance meus objetivos, por todos os valores e educação transmitidos e por ser a minha fortaleza nos momentos mais difíceis. Tudo que faço é por vocês! A minha irmã, Raiane, por me ensinar o quão bom é ter alguém para dividir os momentos e por ser uma certeza de porto seguro independente do que aconteça. A minha orientadora, Prof.ª Dr.ª Ivone Antônia de Souza, por ter me recebido em seu laboratório, investido e acreditado no meu trabalho durante todo trajeto percorrido. Pela paciência, boa vontade, carinho e amizade no decorrer desse tempo, meu imensurável agradecimento. A minha Coorientadora Dr.ª Maria Aparecida Barreto Lopes Seabra, por toda preocupação, esclarecimentos e boa vontade que me auxiliaram na conclusão desse trabalho. Ao Programa de Pós-Graduação em Ciências Farmacêuticas (UFPE), pela oportunidade de realização do mestrado. Aos professores do Programa, que compartilharam comigo o seu conhecimento e a toda equipe de secretária por terem sido sempre tão solícitos em sanar minhas dúvidas e facilitar o percurso. À Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), pelo auxílio financeiro concedido. Aos meus amigos do Laboratório de Farmacologia e Cancerologia Experimental do Departamento de Antibióticos da UFPE que sofreram e sorriram comigo nesta caminhada. Em especial ao grande amigo e companheiro que fiz no decorrer desse trabalho, Rômulo Carlos. À Unileão, à Coordenadora do curso de biomedicina, Ana Ruth Sampaio Granjeiro, à aluna do curso de Biomedicina Ana Karoline e à professora Maria Karollyna do Nascimento, por todo o auxílio oferecido. A colaboração de vocês foi imprescindível para a realização dos testes de atividade antibacteriana. À professora Dr. ª Tassia Campos, pela ajuda e orientação na análise dos dados. Ao professor Fernando Leandro, coordenador no curso de Medicina Veterinária da Universidade Federal Rural de Pernambuco, pela boa vontade e paciência em me auxiliar na leitura e interpretação das lâminas histológicas. Aos técnicos Sandrine Maria e Alex Benício, pela paciência e boa vontade em me transmitir seus conhecimentos e me auxiliar na confecção das lâminas histológicas. A toda minha família que torce diariamente pelo meu sucesso. Que sempre foram grandes incentivadores dos meus sonhos, obrigada pela paciência, pelos incansáveis telefonemas e todas as orações intencionadas a mim ao longo desta caminhada. Amo vocês! Aos amigos que sempre estiveram presentes e a aqueles que fiz no decorrer desse percurso, obrigada por todos os risos e momentos de descontração. Ao meu grande amor, companheira, confidente e apoio de todas as horas e momentos, Maira Rodrigues. Obrigada, por ser sempre a minha grande motivadora, meu aconchego e minha calma. RESUMO Jatropha mollissima (Pohl) Baill. da família Euphorbiaceae é encontrada na Caatinga Brasileira e amplamente distribuída na região nordeste, porém com poucos estudos de seu potencial terapêutico. O objetivo dessa pesquisa foi realizar o perfil fitoquímico do extrato etanólico bruto das folhas de Jatropha mollíssima (EEJM) e analisar as suas atividades toxicológicas, antitumoral e antibacteriana. A coleta do vegetal foi realizada na cidade de Caruaru-PE e encaminhada a Universidade Federal de Pernambuco (UFPE) para identificação e obtenção do extrato etanólico. O perfil fitoquímico foi realizado através de Prospecção preliminar e Cromatografia de Camada Delgada (CCD). O protocolo para realização da toxicidade aguda foi baseado na Metodologia da Organisation for Economic Co-operation and Development (OECD) 423. Avaliação da Atividade antitumoral foi realizada utilizando-se camundongos albinos Swiss fêmeas divididas em 4 grupos com 5 animais (Controle negativo, padrão, e tratamentos 100 mg/kg e 300 mg/kg, respectivamente). Protocolo da avaliação da atividade antitumoral foi realizado de acordo com Geran e colaboradores (1972) com algumas modificações. Nos testes de atividade antibacteriana e modulatória foi empregada a metodologia de microdiluição de acordo com National Comittee For Clinical Laboratory Standards (NCCLS) com finalidade de determinar a Concentração Inibitória Mínima (CIM). Para atividade modulatória foram utilizados os antibióticos Gentamicina e a Amicacina. Em relação ao preparo do inóculo foram utilizadas as linhagens padrões e resistentes de Staphylococcus aureus e Escherichia coli. Os resultados obtidos através das análises fitoquímicas demonstraram a presença de alguns metabólitos secundários tais como, catequinas, cumarinas, esteróides livres, flavonas, flavonóis, xantonas, flavanonas, glicosideos, taninos condensados ou catéquicos e terpenos. De acordo com os resultados do ensaio de toxicidade aguda, o extrato foi classificado na categoria 5 (como referência o Globally Harmonised System – GHS) e sua Dose Letal50 (DL50) foi estipulada como ≥5000 mg, entretanto, histologicamente foi verificado sinais de hepatotoxicidade e nefrotoxicidade. Quanto à atividade antitumoral, as doses de 100 mg/kg e 300 mg/kg demonstraram resultados promissores, com percentual de inibição tumoral de 69,11% e 73,26%, respectivamente, e redução significativa das médias dos pesos dos tumores, histologicamente demostrou sinais de alterações morfológicas nos mesmos. Os resultados da CIM com EEJM revelaram que o mesmo não possui atividade antimicrobiana contra as cepas padrões e resistentes de E.coli e S. aureus mostrando um valor ≥1024 µg/mL, porém quanto à atividade modulatória, os resultados demonstraram sinergismo diante de uma interação física com os antibióticos utilizados, com diminuição significativa da CIM com a gentamicina e amicacina frente a S. aureus e da amicacina frente a E.coli. Todos os resultados obtidos sugerem que o extrato etanólico das folhas de Jatropha mollissima apresentam uma ação promissora na redução de células tumorais e no combate à resistência bacteriana. Mais estudos são necessários para um maior entendimento dos mecanismos de ação das atividades antitumoral e antibacteriana. Palavras-chave: Extratos vegetais. Histologia. Resistência Microbiana a Medicamentos. Sinergia de Substâncias Bioativas. Testes de Drogas Anticâncer. Testes de Toxicidade Aguda. ABSTRACT Jatropha mollissima (Pohl) Baill. of the family Euphorbiaceae is found in the Brazilian Caatinga and widely distributed in the northeast region, but with few studies of its therapeutic potential. The objective of this research was to perform the phytochemical profile of the Ethanol Extract of Jatropha mollissima leaves (EEJM) and to analyze its toxicological, antitumoral and antibacterial activities. The collection of the vegetable was carried out in the city of Caruaru- PE and sent to the Federal University of Pernambuco (UFPE) to
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