Avaliação da genotoxicidade de extratos de boldo (Plectranthus ornatus) e graviola (Annona muricata) através do ensaio cometa e do teste de micronúcleo em linfócitos humanos

Detalhes bibliográficos
Autor(a) principal: Rocha, Rodrigo dos Santos
Data de Publicação: 2016
Tipo de documento: Tese
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UEFS
Texto Completo: http://localhost:8080/tede/handle/tede/392
Resumo: The use of vegetables with medicinal purposes contributes significantly to the care of the primary needs of assistance to health due to difficult access of the population to medical and pharmaceutical assistance, the high cost of manufactured medications and a tendency for people to use natural origin products. However, technical and scientific information about most of them, particularly with respect to the genotoxic potential, are still insufficient so that the use can be considered safe. In this context, the aim of this study was to evaluate the genotoxic effects of boldo extracts (Plectranthus ornatus) and soursop (Annona muricata), through the comet assay and micronucleus test in human lymphocytes with cytokinesis blocking (MNCtB). The tests were performed on human lymphocyte cultures exposed to the extracts in three concentrations: 1.000μg/ml, 500μg/ml and 250μg/ml. Hydrogen peroxide (1mM) was used as a positive control in the comet assay and vincristine sulfate (1mM) in MNCtB. As a negative control, 40% ethanol was used in both tests. The processing of the material for the comet assay and MNCtB was done according respectively to the protocols Tice et al. (2000) and Fenech (1993). The comet assay was performed in five repetitions being computed, in each of them, 100 comets in a total of two blades. For MNCtB three repetitions were performed and 1.000 binucleate lymphocytes were computed per slide. The occurrence of DNA damage (comet assay) did not differ between the cell of the cultures exposed to different concentrations of boldo extracts from each other nor compared to the negative control. Higher occurrence of micronuclei (MNCtB) was observed in the cell of the cultures treated with the extract at concentrations of 1.000μg/ml and 500μg/ml when compared to the negative control (p<0.05) and cultures treated with the extract concentration of 250μg/ml. The soursop extracts at concentrations of 1.000μg/ml to 500μg/ml induced damage occurrence to the DNA significantly higher than that observed in the negative control and cultures treated with extract at a concentration of 250μg/ml (p<0.001). The conditions in which this study was performed, the obtained results allow concluding that: 1) boldo extracts are not effective in inducing DNA damage, but depending on the concentration, they present clastogenic and/or aneugenic effects; 2) soursop extracts induce DNA damage under certain concentrations. The results of this study strongly raise the achievement of other addressing the issue, so that the actual genotoxic potential of vegetables analyzed here can be established.
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spelling Cerqueira, Eneida de Moraes Marcílio01167352521http://lattes.cnpq.br/8962123463058368Rocha, Rodrigo dos Santos2016-09-15T21:49:15Z2016-03-28ROCHA, Rodrigo dos Santos. Avaliação da genotoxicidade de extratos de boldo (Plectranthus ornatus) e graviola (Annona muricata) através do ensaio cometa e do teste de micronúcleo em linfócitos humanos. 2016. 85 f. Tese (Doutorado Acadêmico em Biotecnologia)- Universidade Estadual de Feira de Santana, Feira de Santana, 2016.http://localhost:8080/tede/handle/tede/392The use of vegetables with medicinal purposes contributes significantly to the care of the primary needs of assistance to health due to difficult access of the population to medical and pharmaceutical assistance, the high cost of manufactured medications and a tendency for people to use natural origin products. However, technical and scientific information about most of them, particularly with respect to the genotoxic potential, are still insufficient so that the use can be considered safe. In this context, the aim of this study was to evaluate the genotoxic effects of boldo extracts (Plectranthus ornatus) and soursop (Annona muricata), through the comet assay and micronucleus test in human lymphocytes with cytokinesis blocking (MNCtB). The tests were performed on human lymphocyte cultures exposed to the extracts in three concentrations: 1.000μg/ml, 500μg/ml and 250μg/ml. Hydrogen peroxide (1mM) was used as a positive control in the comet assay and vincristine sulfate (1mM) in MNCtB. As a negative control, 40% ethanol was used in both tests. The processing of the material for the comet assay and MNCtB was done according respectively to the protocols Tice et al. (2000) and Fenech (1993). The comet assay was performed in five repetitions being computed, in each of them, 100 comets in a total of two blades. For MNCtB three repetitions were performed and 1.000 binucleate lymphocytes were computed per slide. The occurrence of DNA damage (comet assay) did not differ between the cell of the cultures exposed to different concentrations of boldo extracts from each other nor compared to the negative control. Higher occurrence of micronuclei (MNCtB) was observed in the cell of the cultures treated with the extract at concentrations of 1.000μg/ml and 500μg/ml when compared to the negative control (p<0.05) and cultures treated with the extract concentration of 250μg/ml. The soursop extracts at concentrations of 1.000μg/ml to 500μg/ml induced damage occurrence to the DNA significantly higher than that observed in the negative control and cultures treated with extract at a concentration of 250μg/ml (p<0.001). The conditions in which this study was performed, the obtained results allow concluding that: 1) boldo extracts are not effective in inducing DNA damage, but depending on the concentration, they present clastogenic and/or aneugenic effects; 2) soursop extracts induce DNA damage under certain concentrations. The results of this study strongly raise the achievement of other addressing the issue, so that the actual genotoxic potential of vegetables analyzed here can be established.O uso de vegetais com fins medicinais contribui de forma significativa para o cuidado das necessidades primárias de assistência à saúde devido ao difícil acesso da população à assistência médica e farmacêutica, aos altos custos dos medicamentos industrializados e à uma tendência das pessoas a utilizarem produtos de origem natural. No entanto, informações técnico-científicas acerca da maioria deles, particularmente no que tange ao potencial genotóxico, são ainda insuficientes para que o uso possa ser considerado seguro. Neste contexto, o objetivo deste estudo foi avaliar os efeitos genotóxicos de extratos do boldo (Plectranthus ornatus) e da graviola (Annona muricata), através do ensaio cometa e do teste de micronúcleo em linfócitos humanos com bloqueio da citocinese (MNCtB). Os testes foram feitos em culturas de linfócitos humanos expostas aos extratos em três concentrações: 1.000µg/ml, 500µg/ml e 250µg/ml. Peróxido de hidrogênio (1mM) foi utilizado como controle positivo no ensaio cometa e sulfato de vincristina (1mM) no MNCtB. Como controle negativo foi usado o etanol 40% em ambos os testes. O processamento do material para o ensaio cometa e MNCtB foi feito de acordo, respectivamente, com os protocolos de Tice et al. (2000) e Fenech (1993). O ensaio cometa foi realizado em cinco repetições, sendo computados, em cada uma delas, 100 cometas em um total de duas lâminas. Para o MNCtB foram realizadas três repetições e computados 1.000 linfócitos binucleados por lâmina. A ocorrência de danos ao DNA (ensaio cometa) não diferiu entre as células das culturas expostas às diferentes concentrações dos extratos do boldo entre si e nem quando comparadas ao controle negativo. Maior ocorrência de micronúcleos (MNCtB) foi observada nas células das culturas tratadas com os extratos nas concentrações de 1.000µg/ml e de 500µg/ml quando comparadas ao controle negativo (p<0,05) e às culturas tratadas com o extrato na concentração de 250µg/ml. Os extratos da graviola nas concentrações de 1.000µg/ml e 500µg/ml induziram ocorrência de danos ao DNA significativamente maior do que o observado no controle negativo e nas culturas tratadas com extrato na concentração de 250µg/ml (p<0,001). Nas condições em que este estudo foi realizado, os resultados obtidos permitem concluir que: 1) extratos do boldo não são efetivos em induzir danos ao DNA, mas na dependência da concentração apresentam efeitos clastogênicos e/ou aneugênicos; 2) extratos da graviola induzem danos ao DNA sob determinadas concentrações. Os resultados deste estudo suscitam fortemente a realização de outros abordando o tema, para que o real potencial genotóxico dos vegetais aqui analisados possa ser estabelecido.Submitted by Ricardo Cedraz Duque Moliterno (ricardo.moliterno@uefs.br) on 2016-09-15T21:49:15Z No. of bitstreams: 1 Rodrigo Tese Final PDF.pdf: 7598532 bytes, checksum: 876426158d6c8f445be96952d8f7c27b (MD5)Made available in DSpace on 2016-09-15T21:49:15Z (GMT). No. of bitstreams: 1 Rodrigo Tese Final PDF.pdf: 7598532 bytes, checksum: 876426158d6c8f445be96952d8f7c27b (MD5) Previous issue date: 2016-03-28application/pdfporUniversidade Estadual de Feira de SantanaDoutorado Acadêmico em BiotecnologiaUEFSBrasilDEPARTAMENTO DE CIÊNCIAS BIOLÓGICASBoldoPlectranthus ornatusGraviolaAnnona muricataGenotoxicidadeEnsaio cometaTeste de micronúcleoGenotoxicityComet assayMicronucleus testCIENCIAS BIOLOGICAS::BIOLOGIA GERALAvaliação da genotoxicidade de extratos de boldo (Plectranthus ornatus) e graviola (Annona muricata) através do ensaio cometa e do teste de micronúcleo em linfócitos humanosinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis-68152692297897915436006006005026123383450589282-1634559385931244697info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UEFSinstname:Universidade Estadual de Feira de Santana (UEFS)instacron:UEFSORIGINALRodrigo Tese Final PDF.pdfRodrigo Tese Final PDF.pdfapplication/pdf7598532http://tede2.uefs.br:8080/bitstream/tede/392/2/Rodrigo+Tese++Final+PDF.pdf876426158d6c8f445be96952d8f7c27bMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-82089http://tede2.uefs.br:8080/bitstream/tede/392/1/license.txt7b5ba3d2445355f386edab96125d42b7MD51tede/3922016-09-15 18:49:15.513oai:tede2.uefs.br:8080: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Biblioteca Digital de Teses e Dissertaçõeshttp://tede2.uefs.br:8080/PUBhttp://tede2.uefs.br:8080/oai/requestbcuefs@uefs.br|| bcref@uefs.br||bcuefs@uefs.bropendoar:2016-09-15T21:49:15Biblioteca Digital de Teses e Dissertações da UEFS - Universidade Estadual de Feira de Santana (UEFS)false
dc.title.por.fl_str_mv Avaliação da genotoxicidade de extratos de boldo (Plectranthus ornatus) e graviola (Annona muricata) através do ensaio cometa e do teste de micronúcleo em linfócitos humanos
title Avaliação da genotoxicidade de extratos de boldo (Plectranthus ornatus) e graviola (Annona muricata) através do ensaio cometa e do teste de micronúcleo em linfócitos humanos
spellingShingle Avaliação da genotoxicidade de extratos de boldo (Plectranthus ornatus) e graviola (Annona muricata) através do ensaio cometa e do teste de micronúcleo em linfócitos humanos
Rocha, Rodrigo dos Santos
Boldo
Plectranthus ornatus
Graviola
Annona muricata
Genotoxicidade
Ensaio cometa
Teste de micronúcleo
Genotoxicity
Comet assay
Micronucleus test
CIENCIAS BIOLOGICAS::BIOLOGIA GERAL
title_short Avaliação da genotoxicidade de extratos de boldo (Plectranthus ornatus) e graviola (Annona muricata) através do ensaio cometa e do teste de micronúcleo em linfócitos humanos
title_full Avaliação da genotoxicidade de extratos de boldo (Plectranthus ornatus) e graviola (Annona muricata) através do ensaio cometa e do teste de micronúcleo em linfócitos humanos
title_fullStr Avaliação da genotoxicidade de extratos de boldo (Plectranthus ornatus) e graviola (Annona muricata) através do ensaio cometa e do teste de micronúcleo em linfócitos humanos
title_full_unstemmed Avaliação da genotoxicidade de extratos de boldo (Plectranthus ornatus) e graviola (Annona muricata) através do ensaio cometa e do teste de micronúcleo em linfócitos humanos
title_sort Avaliação da genotoxicidade de extratos de boldo (Plectranthus ornatus) e graviola (Annona muricata) através do ensaio cometa e do teste de micronúcleo em linfócitos humanos
author Rocha, Rodrigo dos Santos
author_facet Rocha, Rodrigo dos Santos
author_role author
dc.contributor.advisor1.fl_str_mv Cerqueira, Eneida de Moraes Marcílio
dc.contributor.authorID.fl_str_mv 01167352521
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/8962123463058368
dc.contributor.author.fl_str_mv Rocha, Rodrigo dos Santos
contributor_str_mv Cerqueira, Eneida de Moraes Marcílio
dc.subject.por.fl_str_mv Boldo
Plectranthus ornatus
Graviola
Annona muricata
Genotoxicidade
Ensaio cometa
Teste de micronúcleo
topic Boldo
Plectranthus ornatus
Graviola
Annona muricata
Genotoxicidade
Ensaio cometa
Teste de micronúcleo
Genotoxicity
Comet assay
Micronucleus test
CIENCIAS BIOLOGICAS::BIOLOGIA GERAL
dc.subject.eng.fl_str_mv Genotoxicity
Comet assay
Micronucleus test
dc.subject.cnpq.fl_str_mv CIENCIAS BIOLOGICAS::BIOLOGIA GERAL
description The use of vegetables with medicinal purposes contributes significantly to the care of the primary needs of assistance to health due to difficult access of the population to medical and pharmaceutical assistance, the high cost of manufactured medications and a tendency for people to use natural origin products. However, technical and scientific information about most of them, particularly with respect to the genotoxic potential, are still insufficient so that the use can be considered safe. In this context, the aim of this study was to evaluate the genotoxic effects of boldo extracts (Plectranthus ornatus) and soursop (Annona muricata), through the comet assay and micronucleus test in human lymphocytes with cytokinesis blocking (MNCtB). The tests were performed on human lymphocyte cultures exposed to the extracts in three concentrations: 1.000μg/ml, 500μg/ml and 250μg/ml. Hydrogen peroxide (1mM) was used as a positive control in the comet assay and vincristine sulfate (1mM) in MNCtB. As a negative control, 40% ethanol was used in both tests. The processing of the material for the comet assay and MNCtB was done according respectively to the protocols Tice et al. (2000) and Fenech (1993). The comet assay was performed in five repetitions being computed, in each of them, 100 comets in a total of two blades. For MNCtB three repetitions were performed and 1.000 binucleate lymphocytes were computed per slide. The occurrence of DNA damage (comet assay) did not differ between the cell of the cultures exposed to different concentrations of boldo extracts from each other nor compared to the negative control. Higher occurrence of micronuclei (MNCtB) was observed in the cell of the cultures treated with the extract at concentrations of 1.000μg/ml and 500μg/ml when compared to the negative control (p<0.05) and cultures treated with the extract concentration of 250μg/ml. The soursop extracts at concentrations of 1.000μg/ml to 500μg/ml induced damage occurrence to the DNA significantly higher than that observed in the negative control and cultures treated with extract at a concentration of 250μg/ml (p<0.001). The conditions in which this study was performed, the obtained results allow concluding that: 1) boldo extracts are not effective in inducing DNA damage, but depending on the concentration, they present clastogenic and/or aneugenic effects; 2) soursop extracts induce DNA damage under certain concentrations. The results of this study strongly raise the achievement of other addressing the issue, so that the actual genotoxic potential of vegetables analyzed here can be established.
publishDate 2016
dc.date.accessioned.fl_str_mv 2016-09-15T21:49:15Z
dc.date.issued.fl_str_mv 2016-03-28
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dc.identifier.citation.fl_str_mv ROCHA, Rodrigo dos Santos. Avaliação da genotoxicidade de extratos de boldo (Plectranthus ornatus) e graviola (Annona muricata) através do ensaio cometa e do teste de micronúcleo em linfócitos humanos. 2016. 85 f. Tese (Doutorado Acadêmico em Biotecnologia)- Universidade Estadual de Feira de Santana, Feira de Santana, 2016.
dc.identifier.uri.fl_str_mv http://localhost:8080/tede/handle/tede/392
identifier_str_mv ROCHA, Rodrigo dos Santos. Avaliação da genotoxicidade de extratos de boldo (Plectranthus ornatus) e graviola (Annona muricata) através do ensaio cometa e do teste de micronúcleo em linfócitos humanos. 2016. 85 f. Tese (Doutorado Acadêmico em Biotecnologia)- Universidade Estadual de Feira de Santana, Feira de Santana, 2016.
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