Atividade citotóxica das frações, subfrações e annonacinona obtidos de sementes de Annona muricata L

Detalhes bibliográficos
Ano de defesa: 2017
Autor(a) principal: Rocha, Gabriel Gusmão Grisi
Orientador(a): Moraes Filho, Manoel Odorico de
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Não Informado pela instituição
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
Link de acesso: http://www.repositorio.ufc.br/handle/riufc/22014
Resumo: The use of medicinal plants has been evidenced since the beginning to relieve and treat diseases and its use has grown over the years, constituting an important source in the therapeutic arsenal. The species Annona muricata, popularly known as soursop, is widely used as a traditional drug and studies conducted with its bioactive secondary metabolites have shown remarkable antitumor activity related to acetogenins. In this context, the present study aimed the bioguided fractionation of the acetonic extract of the A. muricata seeds to prospect for molecules with cytotoxic potential in human tumor cell lines, resulting in the isolation of an acetogenin called annonacinone. The study is a pioneer in the evaluation of the cytotoxic activity of the molecule. All fractions, subfractions and annonacinone presented cytotoxic potential in the tumor lines tested. The annonacinone showed IC50 values ranging from 3.7 μM to 0.19 μM in colon tumor cells (SW620) and glioblastoma (SF-295), respectively after 72 hours of incubation. The cytotoxic profile at different periods of incubation was performed on SF-295 cells and lung cells (NCI-H460). The cytotoxic effect on SF-295 cells was observed only after 72 hours of incubation, whereas in NCI-H460 cells, this effect was observed at 48 hours with IC50 of 0.21 μM and maintained after 72 hours of incubation. The temporal analysis of the cytotoxicity of annonacinone in NCI-H460 cells was evaluated by two different assays, MTT and SRB, which showed differences in the values, suggesting that the SRB assay is more effective for this kind of substance class because it depends on the protein content without interference of the compound. Annonacinone showed a decrease in cell density at all concentrations tested on NCI-H460 cells after 48 hours of incubation, although after this time of treatment it was not possible to evaluate DNA damage, cell cycle progression and cell death pattern. In an real-time analysis of the proliferation and viability, the annonacinone showed inhibition of the cellular growth in the tested concentrations and beginning of decline of the cellular index after 56 hours of treatment. It is concluded that acetogenin annonacinone presents evident cytotoxic activity in vitro, with greater effect in NCI-H460 cells, however, other tests should be performed with longer treatment times for a better evaluation of its biological activity and therapeutic potential of this molecule.
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spelling Rocha, Gabriel Gusmão GrisiMoraes Filho, Manoel Odorico de2017-02-14T15:43:19Z2017-02-14T15:43:19Z2017-01-26ROCHA, G. G. G. Atividade citotóxica das frações, subfrações e annonacinona obtidos de sementes de Annona muricata L. 2017. 91 f. Dissertação (Mestrado em Farmacologia) - Faculdade de Farmácia, Odontologia e Enfermagem, Universidade Federal do Ceará, Fortaleza, 2017.http://www.repositorio.ufc.br/handle/riufc/22014The use of medicinal plants has been evidenced since the beginning to relieve and treat diseases and its use has grown over the years, constituting an important source in the therapeutic arsenal. The species Annona muricata, popularly known as soursop, is widely used as a traditional drug and studies conducted with its bioactive secondary metabolites have shown remarkable antitumor activity related to acetogenins. In this context, the present study aimed the bioguided fractionation of the acetonic extract of the A. muricata seeds to prospect for molecules with cytotoxic potential in human tumor cell lines, resulting in the isolation of an acetogenin called annonacinone. The study is a pioneer in the evaluation of the cytotoxic activity of the molecule. All fractions, subfractions and annonacinone presented cytotoxic potential in the tumor lines tested. The annonacinone showed IC50 values ranging from 3.7 μM to 0.19 μM in colon tumor cells (SW620) and glioblastoma (SF-295), respectively after 72 hours of incubation. The cytotoxic profile at different periods of incubation was performed on SF-295 cells and lung cells (NCI-H460). The cytotoxic effect on SF-295 cells was observed only after 72 hours of incubation, whereas in NCI-H460 cells, this effect was observed at 48 hours with IC50 of 0.21 μM and maintained after 72 hours of incubation. The temporal analysis of the cytotoxicity of annonacinone in NCI-H460 cells was evaluated by two different assays, MTT and SRB, which showed differences in the values, suggesting that the SRB assay is more effective for this kind of substance class because it depends on the protein content without interference of the compound. Annonacinone showed a decrease in cell density at all concentrations tested on NCI-H460 cells after 48 hours of incubation, although after this time of treatment it was not possible to evaluate DNA damage, cell cycle progression and cell death pattern. In an real-time analysis of the proliferation and viability, the annonacinone showed inhibition of the cellular growth in the tested concentrations and beginning of decline of the cellular index after 56 hours of treatment. It is concluded that acetogenin annonacinone presents evident cytotoxic activity in vitro, with greater effect in NCI-H460 cells, however, other tests should be performed with longer treatment times for a better evaluation of its biological activity and therapeutic potential of this molecule.A utilização de plantas medicinais tem sido evidenciada desde os primórdios para aliviar e tratar doenças e sua utilização tem crescido com o passar dos anos, constituindo uma importante fonte no arsenal terapêutico. A espécie Annona muricata, conhecida popularmente como gravioleira, possui ampla utilização como medicamento tradicional e estudos conduzidos com seus metabólitos secundários bioativos vêm apresentando notável atividade antitumoral ligada às acetogeninas. Nesse contexto o presente estudo objetivou o fracionamento bioguiado do extrato acetônico das sementes de A. muricata para prospecção de moléculas com potencial citotóxico em linhagens celulares tumorais humanas, resultando no isolamento de uma acetogenina denominada annonacinona. O estudo é pioneiro na avaliação da atividade citotóxica da molécula. Todas as frações, subfrações e annonacinona apresentaram potencial citotóxico nas linhagens tumorais testadas. A annonacinona apresentou valores de CI50 que variaram de 3,7 µM a 0,19 µM em células tumorais de cólon (SW620) e glioblastoma (SF-295), respectivamente após 72 horas de incubação. O perfil citotóxico em diferentes períodos de incubação foi realizado em células SF-295 e células de pulmão (NCI-H460). O efeito citotóxico em células SF-295 foi observado apenas após 72 horas de incubação, enquanto que em células NCI-H460, esse efeito foi observado em 48 horas com CI50 de 0,21 µM e mantendo-se após 72 horas de incubação. A análise temporal da citotoxicidade da annonacinona em células NCI-H460 foi avaliada por dois ensaios distintos, MTT e SRB que apresentaram divergências nos valores, sugerindo que o ensaio do SRB seja mais efetivo para esse tipo de classe de substâncias por depender do conteúdo de proteínas sem interferência do composto. A annonacinona apresentou diminuição da densidade celular em todas as concentrações testadas em células NCI-H460 após 48 horas de incubação, embora após esse tempo de tratamento não tenha sido possível avaliar dano de DNA, progressão do ciclo celular e padrão de morte celular. Em análise sobre a proliferação e viabilidade em tempo real, a annonacinona apresentou inibição do crescimento celular nas concentrações testadas e início de declínio do índex celular após 56 horas de tratamento. Conclui-se que a acetogenina annonacinona apresenta evidente atividade citotóxica in vitro, com maior efeito em células NCI-H460, no entanto, outros testes devem ser realizados com tempos de tratamentos maiores para melhor avaliação da sua atividade biológica e potencial terapêutico desta molécula.AcetogeninasCitotoxicidade ImunológicaAnnonaAtividade citotóxica das frações, subfrações e annonacinona obtidos de sementes de Annona muricata LCytotoxic activity of fraction, subfractions and annonacinone obtained by seeds of Annona muricata L.info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccessORIGINAL2017_dis_gggrocha.pdf2017_dis_gggrocha.pdfapplication/pdf2091365http://repositorio.ufc.br/bitstream/riufc/22014/1/2017_dis_gggrocha.pdf747a32d7ce7e39158d50adf61b230f08MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufc.br/bitstream/riufc/22014/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52riufc/220142019-10-18 08:54:42.548oai:repositorio.ufc.br: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Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2019-10-18T11:54:42Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.pt_BR.fl_str_mv Atividade citotóxica das frações, subfrações e annonacinona obtidos de sementes de Annona muricata L
dc.title.en.pt_BR.fl_str_mv Cytotoxic activity of fraction, subfractions and annonacinone obtained by seeds of Annona muricata L.
title Atividade citotóxica das frações, subfrações e annonacinona obtidos de sementes de Annona muricata L
spellingShingle Atividade citotóxica das frações, subfrações e annonacinona obtidos de sementes de Annona muricata L
Rocha, Gabriel Gusmão Grisi
Acetogeninas
Citotoxicidade Imunológica
Annona
title_short Atividade citotóxica das frações, subfrações e annonacinona obtidos de sementes de Annona muricata L
title_full Atividade citotóxica das frações, subfrações e annonacinona obtidos de sementes de Annona muricata L
title_fullStr Atividade citotóxica das frações, subfrações e annonacinona obtidos de sementes de Annona muricata L
title_full_unstemmed Atividade citotóxica das frações, subfrações e annonacinona obtidos de sementes de Annona muricata L
title_sort Atividade citotóxica das frações, subfrações e annonacinona obtidos de sementes de Annona muricata L
author Rocha, Gabriel Gusmão Grisi
author_facet Rocha, Gabriel Gusmão Grisi
author_role author
dc.contributor.author.fl_str_mv Rocha, Gabriel Gusmão Grisi
dc.contributor.advisor1.fl_str_mv Moraes Filho, Manoel Odorico de
contributor_str_mv Moraes Filho, Manoel Odorico de
dc.subject.por.fl_str_mv Acetogeninas
Citotoxicidade Imunológica
Annona
topic Acetogeninas
Citotoxicidade Imunológica
Annona
description The use of medicinal plants has been evidenced since the beginning to relieve and treat diseases and its use has grown over the years, constituting an important source in the therapeutic arsenal. The species Annona muricata, popularly known as soursop, is widely used as a traditional drug and studies conducted with its bioactive secondary metabolites have shown remarkable antitumor activity related to acetogenins. In this context, the present study aimed the bioguided fractionation of the acetonic extract of the A. muricata seeds to prospect for molecules with cytotoxic potential in human tumor cell lines, resulting in the isolation of an acetogenin called annonacinone. The study is a pioneer in the evaluation of the cytotoxic activity of the molecule. All fractions, subfractions and annonacinone presented cytotoxic potential in the tumor lines tested. The annonacinone showed IC50 values ranging from 3.7 μM to 0.19 μM in colon tumor cells (SW620) and glioblastoma (SF-295), respectively after 72 hours of incubation. The cytotoxic profile at different periods of incubation was performed on SF-295 cells and lung cells (NCI-H460). The cytotoxic effect on SF-295 cells was observed only after 72 hours of incubation, whereas in NCI-H460 cells, this effect was observed at 48 hours with IC50 of 0.21 μM and maintained after 72 hours of incubation. The temporal analysis of the cytotoxicity of annonacinone in NCI-H460 cells was evaluated by two different assays, MTT and SRB, which showed differences in the values, suggesting that the SRB assay is more effective for this kind of substance class because it depends on the protein content without interference of the compound. Annonacinone showed a decrease in cell density at all concentrations tested on NCI-H460 cells after 48 hours of incubation, although after this time of treatment it was not possible to evaluate DNA damage, cell cycle progression and cell death pattern. In an real-time analysis of the proliferation and viability, the annonacinone showed inhibition of the cellular growth in the tested concentrations and beginning of decline of the cellular index after 56 hours of treatment. It is concluded that acetogenin annonacinone presents evident cytotoxic activity in vitro, with greater effect in NCI-H460 cells, however, other tests should be performed with longer treatment times for a better evaluation of its biological activity and therapeutic potential of this molecule.
publishDate 2017
dc.date.accessioned.fl_str_mv 2017-02-14T15:43:19Z
dc.date.available.fl_str_mv 2017-02-14T15:43:19Z
dc.date.issued.fl_str_mv 2017-01-26
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dc.identifier.citation.fl_str_mv ROCHA, G. G. G. Atividade citotóxica das frações, subfrações e annonacinona obtidos de sementes de Annona muricata L. 2017. 91 f. Dissertação (Mestrado em Farmacologia) - Faculdade de Farmácia, Odontologia e Enfermagem, Universidade Federal do Ceará, Fortaleza, 2017.
dc.identifier.uri.fl_str_mv http://www.repositorio.ufc.br/handle/riufc/22014
identifier_str_mv ROCHA, G. G. G. Atividade citotóxica das frações, subfrações e annonacinona obtidos de sementes de Annona muricata L. 2017. 91 f. Dissertação (Mestrado em Farmacologia) - Faculdade de Farmácia, Odontologia e Enfermagem, Universidade Federal do Ceará, Fortaleza, 2017.
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