Toxicidade de Eugenia uniflora L. (myrtaceae) em modelos de Drosophila melanogaster e células sanguíneas humanas

Detalhes bibliográficos
Autor(a) principal: Cunha, Francisco Assis Bezerra da
Data de Publicação: 2015
Tipo de documento: Tese
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
dARK ID: ark:/26339/001300000rgbn
Texto Completo: http://repositorio.ufsm.br/handle/1/17593
Resumo: The increasing use of natural products has led to the imperative need of investigating the toxicological potential of secondary plant metabolites. In this context, toxicological studies were carried on Eugenia uniflora (family: Myrtaceae), a species used in Ceara state, Brazil, for medicinal purpose. Particularly, this study investigated the toxicity of E. uniflora leaf essential oil in Drosophila melanogaster as well as the toxicity of ethanolic extract of E. uniflora in human cells. The phytochemical profile of the essential oil analyzed by GC-MS and GC-FID showed curzerene (48.06%), γ-elemene (13.49%) atractilone (11.78%) and trans- β-elemenone (8.94%) as the major constituents. Polyphenolic constituents of E. uniflora ethanolic extract analysed by high performance liquid chromatography (HPLC) revealed the presence of ellagic acid (1.19%), cyanidin (0.56%), quercetin (1.58%), quercitrin (1.34%), isoquercitrin (1.01%) and Luteolin (1.01%) as the major components. The extract at the concentrations tested (1-480 μg/mL) showed no cytotoxic effect to leukocytes and erythrocytes. In addition, the extract did not have any genotoxic effect, suggesting that the extract can be consumed safely at relatively high concentrations. The extract exhibited lower DPPH radical scavenging activity in comparison to ascorbic acid, but strongly inhibited (30- 480 μg/mL) lipid Fe2+ (10 μM)-induced lipid peroxidation (LPO) in rat brain and liver homogenates The results obtained with E. uniflora leaf essential oil indicate that it induces mortality in D. melanogaster (LC50 = 5.56 μg/mL) and locomotor deficit after 12 h. Based on this observation, the flies were exposed to 3 μg/mL of essential oil for 3, 6 and 12 h. Exposure of flies to 3 h caused a significant increase in reactive species production which remained stable from 6 to 12 h. There was an increase in TBARS formation following exposure of flies to E. uniflora leaf essential oil after 6 and 12 h, and this was associated with a significant increase in GST and SOD activities. A significant increase in the expression of NQO-1 was noted after 3 h of exposure, and this was associated with a significant increase in the protein level of HSP70 after 12 h. Basal levels of Nrf2 transcription factor remained unchanged. Co-exposure of the essential oil with Paraquat (20 mM) and Fe 2+ (10 mM) increased the mortality (104.4% and 98.82%) compared to the control. Similarly, Paraquat and Fe2+ increased the locomotor deficit (35.94% and 103.1%) respectively. These results indicate the toxic effects of E. uniflora leaf essential oil on D. melanogaster and points oxidative stress as an importante mechanism of toxicity. Nevertheless, further studies should be conducted to better understand the mechanism(s) underlying its toxicity.
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spelling Toxicidade de Eugenia uniflora L. (myrtaceae) em modelos de Drosophila melanogaster e células sanguíneas humanasToxicity of Eugenia uniflora L. (myrtaceae) in Drosophila melanogaster and human blood cellsSinalização celularGeotaxia negativaCromatografiaCell signalingNegative geotaxiaChromatographyCNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICAThe increasing use of natural products has led to the imperative need of investigating the toxicological potential of secondary plant metabolites. In this context, toxicological studies were carried on Eugenia uniflora (family: Myrtaceae), a species used in Ceara state, Brazil, for medicinal purpose. Particularly, this study investigated the toxicity of E. uniflora leaf essential oil in Drosophila melanogaster as well as the toxicity of ethanolic extract of E. uniflora in human cells. The phytochemical profile of the essential oil analyzed by GC-MS and GC-FID showed curzerene (48.06%), γ-elemene (13.49%) atractilone (11.78%) and trans- β-elemenone (8.94%) as the major constituents. Polyphenolic constituents of E. uniflora ethanolic extract analysed by high performance liquid chromatography (HPLC) revealed the presence of ellagic acid (1.19%), cyanidin (0.56%), quercetin (1.58%), quercitrin (1.34%), isoquercitrin (1.01%) and Luteolin (1.01%) as the major components. The extract at the concentrations tested (1-480 μg/mL) showed no cytotoxic effect to leukocytes and erythrocytes. In addition, the extract did not have any genotoxic effect, suggesting that the extract can be consumed safely at relatively high concentrations. The extract exhibited lower DPPH radical scavenging activity in comparison to ascorbic acid, but strongly inhibited (30- 480 μg/mL) lipid Fe2+ (10 μM)-induced lipid peroxidation (LPO) in rat brain and liver homogenates The results obtained with E. uniflora leaf essential oil indicate that it induces mortality in D. melanogaster (LC50 = 5.56 μg/mL) and locomotor deficit after 12 h. Based on this observation, the flies were exposed to 3 μg/mL of essential oil for 3, 6 and 12 h. Exposure of flies to 3 h caused a significant increase in reactive species production which remained stable from 6 to 12 h. There was an increase in TBARS formation following exposure of flies to E. uniflora leaf essential oil after 6 and 12 h, and this was associated with a significant increase in GST and SOD activities. A significant increase in the expression of NQO-1 was noted after 3 h of exposure, and this was associated with a significant increase in the protein level of HSP70 after 12 h. Basal levels of Nrf2 transcription factor remained unchanged. Co-exposure of the essential oil with Paraquat (20 mM) and Fe 2+ (10 mM) increased the mortality (104.4% and 98.82%) compared to the control. Similarly, Paraquat and Fe2+ increased the locomotor deficit (35.94% and 103.1%) respectively. These results indicate the toxic effects of E. uniflora leaf essential oil on D. melanogaster and points oxidative stress as an importante mechanism of toxicity. Nevertheless, further studies should be conducted to better understand the mechanism(s) underlying its toxicity.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESA crescente utilização de produtos naturais torna imperativa a investigação do potencial toxicológico dos metabólitos secundários de plantas. Neste contexto insere-se o estudo toxicológico de Eugenia uniflora, espécie pertencente à família Myrtaceae, a maior família das Angiospermas. Este trabalho investigou a toxicidade do óleo essencial desta planta em modelo in vivo de Drosophila melanogaster e do extrato etanólico em células sanguíneas humanas. O perfil fitoquímico do óleo essencial foi analisado por GC-MS e GC-FID e apresentou como constituintes majoritários: curzereno (48,06%), γ-elemeno (13,49%), atractilone (11,78%) e trans-β-elemenone (8,94%). Os fitoconstituintes majoritários do extrato etanólico, identificados por HPLC, foram: ácido elágico (1,19%), cianidina (0,56%), quercetina (1,58%), quercitrina (1,34%), isoquercitrina (1,01%) e luteolina (1,01%). O extrato, nas concentrações (1-480 μg∕mL) não apresentou citotoxicidade pelos modelos de viabilidade celular em leucócitos humanos e de fragilidade osmótica em eritrócitos, nem genotoxicidade avaliada pelo Ensaio Cometa. Tendo apresentado uma atividade concentração dependente em modelo de DPPH, quando comparado ao ácido ascórbico. A sua atividade antioxidante em condições basais, nas concentrações de (30-480 μg/mL) inibiu a formação de TBARS. Com inibição máxima (30 μg/mL) no cérebro e (120 μg/mL) no fígado. O extrato também atenuou a formação de TBARS induzida por Fe2+ (10 μM) nas concentrações de (120 μg/mL) para o cérebro e (240 μg/mL) para o fígado. O óleo essencial induziu mortalidade em D. melanogaster com CL50 (5,56 μg∕mL) em 12 h e déficit na capacidade locomotora. Foi escolhida uma concentração subletal de (3 μg∕mL) em intervalos de tempo de 3, 6 e 12 horas para realização dos testes. Em paralelo, as moscas também apresentaram sinais de estresse oxidativo, incluindo formação de ERO’s em 3 h de exposição o que se manteve em 6 e 12 h. Apresentando aumento nos níveis de TBARS em 6 e 12 h de exposição. Um aumento na atividade da enzima GST às 6 e 12 h e da SOD às 12 h foram significantes. Apresentou, também, um aumento significativo na expressão de NQO-1 a 3 h de exposição. O nível da proteína HSP70 apresentou um aumento significativo as 12 h. Os níveis do fator de transcrição Nrf2 permaneceram inalterados. A co-exposição do óleo essencial com o Paraquat (20 mM) e Ferro (10 mM), aumentou a toxicidade em (104,4% e 98,82%) respectivamente. De forma similar, Paraquat e Ferro aumentou, respectivamente, o déficit locomotor em (35,94% e 103,1%) em relação ao controle. Em resumo, o extrato etanólico não apresentou toxicidade nas concentrações e modelos testados e o óleo essencial apresentou toxicidade em D. melanogaster, tendo o estresse oxidativo como importante mecanismo de ação. A partir dos estudos realizados, sugere-se uma potencial aplicação do óleo essencial como inseticida de origem natural. Novos estudos necessitam ser realizados para melhor compreender os mecanismos toxicológicos envolvidos.Universidade Federal de Santa MariaBrasilBioquímicaUFSMPrograma de Pós-Graduação em Ciências Biológicas: Bioquímica ToxicológicaCentro de Ciências Naturais e ExatasFranco, Jeferson Luishttp://lattes.cnpq.br/1680065573338339Rubin, Maribel Antonellohttp://lattes.cnpq.br/7237734243628134Oliveira, Sara Marchesan dehttp://lattes.cnpq.br/6574555059806902Barros, Adriana Rolim Camposhttp://lattes.cnpq.br/3791336333658295Menezes, Irwin Rose Alencar dehttp://lattes.cnpq.br/6310868104861653Cunha, Francisco Assis Bezerra da2019-07-29T19:19:27Z2019-07-29T19:19:27Z2015-12-17info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/17593ark:/26339/001300000rgbnporAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2019-07-30T06:02:33Zoai:repositorio.ufsm.br:1/17593Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2019-07-30T06:02:33Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv Toxicidade de Eugenia uniflora L. (myrtaceae) em modelos de Drosophila melanogaster e células sanguíneas humanas
Toxicity of Eugenia uniflora L. (myrtaceae) in Drosophila melanogaster and human blood cells
title Toxicidade de Eugenia uniflora L. (myrtaceae) em modelos de Drosophila melanogaster e células sanguíneas humanas
spellingShingle Toxicidade de Eugenia uniflora L. (myrtaceae) em modelos de Drosophila melanogaster e células sanguíneas humanas
Cunha, Francisco Assis Bezerra da
Sinalização celular
Geotaxia negativa
Cromatografia
Cell signaling
Negative geotaxia
Chromatography
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
title_short Toxicidade de Eugenia uniflora L. (myrtaceae) em modelos de Drosophila melanogaster e células sanguíneas humanas
title_full Toxicidade de Eugenia uniflora L. (myrtaceae) em modelos de Drosophila melanogaster e células sanguíneas humanas
title_fullStr Toxicidade de Eugenia uniflora L. (myrtaceae) em modelos de Drosophila melanogaster e células sanguíneas humanas
title_full_unstemmed Toxicidade de Eugenia uniflora L. (myrtaceae) em modelos de Drosophila melanogaster e células sanguíneas humanas
title_sort Toxicidade de Eugenia uniflora L. (myrtaceae) em modelos de Drosophila melanogaster e células sanguíneas humanas
author Cunha, Francisco Assis Bezerra da
author_facet Cunha, Francisco Assis Bezerra da
author_role author
dc.contributor.none.fl_str_mv Franco, Jeferson Luis
http://lattes.cnpq.br/1680065573338339
Rubin, Maribel Antonello
http://lattes.cnpq.br/7237734243628134
Oliveira, Sara Marchesan de
http://lattes.cnpq.br/6574555059806902
Barros, Adriana Rolim Campos
http://lattes.cnpq.br/3791336333658295
Menezes, Irwin Rose Alencar de
http://lattes.cnpq.br/6310868104861653
dc.contributor.author.fl_str_mv Cunha, Francisco Assis Bezerra da
dc.subject.por.fl_str_mv Sinalização celular
Geotaxia negativa
Cromatografia
Cell signaling
Negative geotaxia
Chromatography
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
topic Sinalização celular
Geotaxia negativa
Cromatografia
Cell signaling
Negative geotaxia
Chromatography
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
description The increasing use of natural products has led to the imperative need of investigating the toxicological potential of secondary plant metabolites. In this context, toxicological studies were carried on Eugenia uniflora (family: Myrtaceae), a species used in Ceara state, Brazil, for medicinal purpose. Particularly, this study investigated the toxicity of E. uniflora leaf essential oil in Drosophila melanogaster as well as the toxicity of ethanolic extract of E. uniflora in human cells. The phytochemical profile of the essential oil analyzed by GC-MS and GC-FID showed curzerene (48.06%), γ-elemene (13.49%) atractilone (11.78%) and trans- β-elemenone (8.94%) as the major constituents. Polyphenolic constituents of E. uniflora ethanolic extract analysed by high performance liquid chromatography (HPLC) revealed the presence of ellagic acid (1.19%), cyanidin (0.56%), quercetin (1.58%), quercitrin (1.34%), isoquercitrin (1.01%) and Luteolin (1.01%) as the major components. The extract at the concentrations tested (1-480 μg/mL) showed no cytotoxic effect to leukocytes and erythrocytes. In addition, the extract did not have any genotoxic effect, suggesting that the extract can be consumed safely at relatively high concentrations. The extract exhibited lower DPPH radical scavenging activity in comparison to ascorbic acid, but strongly inhibited (30- 480 μg/mL) lipid Fe2+ (10 μM)-induced lipid peroxidation (LPO) in rat brain and liver homogenates The results obtained with E. uniflora leaf essential oil indicate that it induces mortality in D. melanogaster (LC50 = 5.56 μg/mL) and locomotor deficit after 12 h. Based on this observation, the flies were exposed to 3 μg/mL of essential oil for 3, 6 and 12 h. Exposure of flies to 3 h caused a significant increase in reactive species production which remained stable from 6 to 12 h. There was an increase in TBARS formation following exposure of flies to E. uniflora leaf essential oil after 6 and 12 h, and this was associated with a significant increase in GST and SOD activities. A significant increase in the expression of NQO-1 was noted after 3 h of exposure, and this was associated with a significant increase in the protein level of HSP70 after 12 h. Basal levels of Nrf2 transcription factor remained unchanged. Co-exposure of the essential oil with Paraquat (20 mM) and Fe 2+ (10 mM) increased the mortality (104.4% and 98.82%) compared to the control. Similarly, Paraquat and Fe2+ increased the locomotor deficit (35.94% and 103.1%) respectively. These results indicate the toxic effects of E. uniflora leaf essential oil on D. melanogaster and points oxidative stress as an importante mechanism of toxicity. Nevertheless, further studies should be conducted to better understand the mechanism(s) underlying its toxicity.
publishDate 2015
dc.date.none.fl_str_mv 2015-12-17
2019-07-29T19:19:27Z
2019-07-29T19:19:27Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://repositorio.ufsm.br/handle/1/17593
dc.identifier.dark.fl_str_mv ark:/26339/001300000rgbn
url http://repositorio.ufsm.br/handle/1/17593
identifier_str_mv ark:/26339/001300000rgbn
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Bioquímica
UFSM
Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica
Centro de Ciências Naturais e Exatas
publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Bioquímica
UFSM
Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica
Centro de Ciências Naturais e Exatas
dc.source.none.fl_str_mv reponame:Manancial - Repositório Digital da UFSM
instname:Universidade Federal de Santa Maria (UFSM)
instacron:UFSM
instname_str Universidade Federal de Santa Maria (UFSM)
instacron_str UFSM
institution UFSM
reponame_str Manancial - Repositório Digital da UFSM
collection Manancial - Repositório Digital da UFSM
repository.name.fl_str_mv Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)
repository.mail.fl_str_mv atendimento.sib@ufsm.br||tedebc@gmail.com
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