Toxicidade de Eugenia uniflora L. (myrtaceae) em modelos de Drosophila melanogaster e células sanguíneas humanas
Autor(a) principal: | |
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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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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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1815172384818200576 |