Estudo do potencial antioxidante da espécie Caryocar coriaceum Wittm. em modelos experimentais in vitro e in vivo
Ano de defesa: | 2016 |
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Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | |
Tipo de documento: | Tese |
Tipo de acesso: | Acesso aberto |
Idioma: | por |
Instituição de defesa: |
Universidade Federal de Santa Maria
Brasil Bioquímica UFSM Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica Centro de Ciências Naturais e Exatas |
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://repositorio.ufsm.br/handle/1/17965 |
Resumo: | Despite the diversity of species found in the National Araripe Forest, Ceará Cariri, there are few studies with Caryocar coriaceumWittm (C. coriaceum), a plant that produces a highly consumed fruit in regional cuisine. The oil extracted from the pulp is also widely used in popular medicine for the treatment of respiratory infections and joint diseases. Studies with oil pulp have consolidated therapeutic properties of species. However, this is the first study with plant leaf extracts and oil from the fruit pulp that assesses markers of oxidative stress and gene expression of antioxidant enzymes in vivo using Drosophila melanogaster. The antioxidant activity of leaf and fruit peel extracts was evaluated in vitro. The major phenolic compounds from extracts, identified by HPLC-DAD, were quercetin, chlorogenic acid and ellagic. In in vitro tests, the extracts exhibited similar and significant DPPH● and OH● scavenging activity, were effective in reducing and chelating iron as well as decreasing lipid peroxidation and production of reactive oxygen species induced by iron and calcium, respectively, in liver homogenate. The extracts did not change the levels O2•– in mitochondria treated with antimycin A. In in vivo study, flies were pretreated with a diet containing aqueous leaf extract (AEL) (1 and 5 mg/g diet) and/or pequi pulp oil (PPO) (1, 5 and 10mg/g diet) for 7 days followed by exposure to paraquat (PQ) (10mM) for 24 hours. A third treatment was performed concomitantly with AEL and PQ (1mM) for 5 days. The analysis of the negative geotaxia was performed to evaluate locomotor ability of flies. Flies homogenate was used to analyze oxidative stress markers specifically activity of catalase (CAT) and glutathione-s-transferase (GST), levels of ROS and peroxidation lipid by levels of TBARS. We evaluated the mRNA expression of antioxidant enzyme genes CAT, superoxide dismutase (SOD), Thioredoxin reductase-1 (Trx R-1), Thioredoxin T (Trx T) and Keap 1 protein by qRT-PCR. Flies exposed to PQ showed a significant impairment in locomotor activity, increase in oxidative stress markers and gene expression of CAT, SOD, Trx R-1, Trx T and Keap 1. The concentrations AEL and OPP, in all treatments, reduced significantly lipid peroxidation levels and reversed locomotor changes induced by exposure to PQ. In both AEL treatments, flies exposed to PQ had a reduction in ROS levels and CAT and GST activities. Flies concomitantly treated with AEL and PQ exhibited a significant decrease in expression of Trx R-1 and Trx T genes, which were increased in the PQ group. Pre-treatment with PPO of 10mg/g diet, we verified a decrease the expression of genes CAT, SOD, Keap 1 and Trx T, which were increased in the group exposed to PQ alone. The data obtained in this study show that the treatment with AEL and PPO reduced oxidative stress markers induced by PQ and suggest that this effect may be related, at least in part, to the antioxidant system modulation. Moreover, these results will collaborate to extend the knowledge about the scientific properties of C. coriaceum as well as point it as a promissing functional food. |
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Estudo do potencial antioxidante da espécie Caryocar coriaceum Wittm. em modelos experimentais in vitro e in vivoEvaluation of the antioxidant potential from Caryocar coriaceum Wittm. in vitro and in vivoCaryocar coriaceum Wittm.PequiEstresse oxidativoEnzimas antioxidantesDrosophila melanogasterOxidative stressAntioxidants enzymesCNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICADespite the diversity of species found in the National Araripe Forest, Ceará Cariri, there are few studies with Caryocar coriaceumWittm (C. coriaceum), a plant that produces a highly consumed fruit in regional cuisine. The oil extracted from the pulp is also widely used in popular medicine for the treatment of respiratory infections and joint diseases. Studies with oil pulp have consolidated therapeutic properties of species. However, this is the first study with plant leaf extracts and oil from the fruit pulp that assesses markers of oxidative stress and gene expression of antioxidant enzymes in vivo using Drosophila melanogaster. The antioxidant activity of leaf and fruit peel extracts was evaluated in vitro. The major phenolic compounds from extracts, identified by HPLC-DAD, were quercetin, chlorogenic acid and ellagic. In in vitro tests, the extracts exhibited similar and significant DPPH● and OH● scavenging activity, were effective in reducing and chelating iron as well as decreasing lipid peroxidation and production of reactive oxygen species induced by iron and calcium, respectively, in liver homogenate. The extracts did not change the levels O2•– in mitochondria treated with antimycin A. In in vivo study, flies were pretreated with a diet containing aqueous leaf extract (AEL) (1 and 5 mg/g diet) and/or pequi pulp oil (PPO) (1, 5 and 10mg/g diet) for 7 days followed by exposure to paraquat (PQ) (10mM) for 24 hours. A third treatment was performed concomitantly with AEL and PQ (1mM) for 5 days. The analysis of the negative geotaxia was performed to evaluate locomotor ability of flies. Flies homogenate was used to analyze oxidative stress markers specifically activity of catalase (CAT) and glutathione-s-transferase (GST), levels of ROS and peroxidation lipid by levels of TBARS. We evaluated the mRNA expression of antioxidant enzyme genes CAT, superoxide dismutase (SOD), Thioredoxin reductase-1 (Trx R-1), Thioredoxin T (Trx T) and Keap 1 protein by qRT-PCR. Flies exposed to PQ showed a significant impairment in locomotor activity, increase in oxidative stress markers and gene expression of CAT, SOD, Trx R-1, Trx T and Keap 1. The concentrations AEL and OPP, in all treatments, reduced significantly lipid peroxidation levels and reversed locomotor changes induced by exposure to PQ. In both AEL treatments, flies exposed to PQ had a reduction in ROS levels and CAT and GST activities. Flies concomitantly treated with AEL and PQ exhibited a significant decrease in expression of Trx R-1 and Trx T genes, which were increased in the PQ group. Pre-treatment with PPO of 10mg/g diet, we verified a decrease the expression of genes CAT, SOD, Keap 1 and Trx T, which were increased in the group exposed to PQ alone. The data obtained in this study show that the treatment with AEL and PPO reduced oxidative stress markers induced by PQ and suggest that this effect may be related, at least in part, to the antioxidant system modulation. Moreover, these results will collaborate to extend the knowledge about the scientific properties of C. coriaceum as well as point it as a promissing functional food.Apesar da diversidade das espécies encontradas na Chapada do Araripe, Cariri cearense, são escassos os estudos com Caryocar coriaceum Wittm. (C. coriaceum), planta que origina um fruto altamente consumido na culinária regional. O óleo extraído da polpa também é bastante usado na medicina popular para o tratamento de infecções respiratórias e doenças articulares. Pesquisas com óleo da polpa vêm consolidando propriedades terapêuticas da espécie. No entanto, esse é o primeiro estudo com extratos de folhas da planta e óleo da polpa da fruta que avalia marcadores de estresse oxidativo e a expressão gênica de enzimas antioxidantes in vivo, usando Drosophila melanogaster. A atividade antioxidante dos extratos de folhas e da casca do fruto também foi avaliada em testes in vitro. Os compostos fenólicos majoritários dos extratos identificados por HPLC-DAD foram quercetina, ácido clorogênico e elágico. Nos testes in vitro, os extratos exibiram semelhante e significante atividade sequestradora de radical DPPH● e OH● ; foram efetivos em reduzir e quelar ferro, assim como, diminuíram a peroxidação lipídica induzida pelo ferro e a produção de espécies reativas induzidas por cálcio em homogenato de fígado de ratos. No entanto, os extratos não modificaram os níveis O2•– em mitocôndrias tratadas com antimicina A. Na etapa do estudo in vivo, as moscas foram pré-tratadas com uma dieta contendo extrato aquoso da folha (EAF) (1 e 5 mg/g dieta) e/ou óleo da polpa de pequi (OPP) (1, 5 e 10mg/g dieta) durante 7 dias, seguida pela exposição ao paraquat (PQ) (10mM) por 24 horas. Um terceiro tratamento foi realizado com EAF concomitantemente a exposição ao PQ (1mM) por 5 dias. A análise da geotaxia negativa foi realizada para avaliar a habilidade locomotora das moscas. O homogenato de moscas foi usado para analisar marcadores de estresse oxidativo especificamente, a determinação da atividade da catalase (CAT) e da Glutationa S-Transferase (GST), os níveis de EROs e da peroxidação lipídica. Foram avaliadas as expressões de mRNA de genes de enzimas antioxidantes CAT, superóxido dismutase (SOD), Tioredoxina redutase-1 (TrxR-1), Tioredoxina T (TrxT) e da proteína Keap 1, através de qRT-PCR. As moscas expostas ao PQ apresentaram um significante prejuízo no comportamento locomotor, um aumento expressivo dos marcadores do estresse oxidativo e na expressão de mRNA para CAT, SOD, Trx R-1, Trx T e Keap 1. As concentrações do EAF e do OPP, em todos os tratamentos, reduziram significativamente os níveis de MDA e reverteram o prejuízo locomotor induzido pela exposição ao PQ. Em ambos os tratamentos com EAF, as moscas expostas ao PQ tiveram uma diminuição dos níveis de EROs e da atividade de CAT e GST. As moscas submetidas ao tratamento concomitante com EAF e PQ apresentaram uma diminuição significativa na expressão dos genes Trx R-1 e Trx T, a qual havia sido aumentada no grupo exposto somente ao PQ. No pré-tratamento com OPP (10mg/g dieta), um efeito similar foi verificado na expressão dos genes CAT, SOD, Keap 1 e Trx T. Os dados obtidos mostram que os tratamentos com o EAF de C. coriaceum e o OPP reduziram os marcadores de estresse oxidativo induzidos por PQ e sugerem que este efeito pode estar relacionado, pelo menos em parte com a modulação do sistema antioxidante. Este estudo colaborou para aumentar o conhecimento sobre as propriedades farmacológicas da espécie C. coriaceum bem como para potencializar seu uso como alimento funcional.Universidade Federal de Santa MariaBrasilBioquímicaUFSMPrograma de Pós-Graduação em Ciências Biológicas: Bioquímica ToxicológicaCentro de Ciências Naturais e ExatasBarbosa, Nilda Berenice de Vargashttp://lattes.cnpq.br/5901511067144019Dalla Corte, Cristiane Lenzhttp://lattes.cnpq.br/5296284169605317Rosemberg, Denis Broockhttp://lattes.cnpq.br/7713953979203056Folmer, Vanderleihttp://lattes.cnpq.br/8135232309980269Menezes, Irwin Rose Alencar dehttp://lattes.cnpq.br/6310868104861653Pereira, Sandra Mara Pimentel Duavy2019-08-19T19:45:45Z2019-08-19T19:45:45Z2016-04-29info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/17965porAttribution-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:UFSM2019-08-20T06:02:18Zoai:repositorio.ufsm.br:1/17965Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2024-07-29T10:27:10.498745Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.none.fl_str_mv |
Estudo do potencial antioxidante da espécie Caryocar coriaceum Wittm. em modelos experimentais in vitro e in vivo Evaluation of the antioxidant potential from Caryocar coriaceum Wittm. in vitro and in vivo |
title |
Estudo do potencial antioxidante da espécie Caryocar coriaceum Wittm. em modelos experimentais in vitro e in vivo |
spellingShingle |
Estudo do potencial antioxidante da espécie Caryocar coriaceum Wittm. em modelos experimentais in vitro e in vivo Pereira, Sandra Mara Pimentel Duavy Caryocar coriaceum Wittm. Pequi Estresse oxidativo Enzimas antioxidantes Drosophila melanogaster Oxidative stress Antioxidants enzymes CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA |
title_short |
Estudo do potencial antioxidante da espécie Caryocar coriaceum Wittm. em modelos experimentais in vitro e in vivo |
title_full |
Estudo do potencial antioxidante da espécie Caryocar coriaceum Wittm. em modelos experimentais in vitro e in vivo |
title_fullStr |
Estudo do potencial antioxidante da espécie Caryocar coriaceum Wittm. em modelos experimentais in vitro e in vivo |
title_full_unstemmed |
Estudo do potencial antioxidante da espécie Caryocar coriaceum Wittm. em modelos experimentais in vitro e in vivo |
title_sort |
Estudo do potencial antioxidante da espécie Caryocar coriaceum Wittm. em modelos experimentais in vitro e in vivo |
author |
Pereira, Sandra Mara Pimentel Duavy |
author_facet |
Pereira, Sandra Mara Pimentel Duavy |
author_role |
author |
dc.contributor.none.fl_str_mv |
Barbosa, Nilda Berenice de Vargas http://lattes.cnpq.br/5901511067144019 Dalla Corte, Cristiane Lenz http://lattes.cnpq.br/5296284169605317 Rosemberg, Denis Broock http://lattes.cnpq.br/7713953979203056 Folmer, Vanderlei http://lattes.cnpq.br/8135232309980269 Menezes, Irwin Rose Alencar de http://lattes.cnpq.br/6310868104861653 |
dc.contributor.author.fl_str_mv |
Pereira, Sandra Mara Pimentel Duavy |
dc.subject.por.fl_str_mv |
Caryocar coriaceum Wittm. Pequi Estresse oxidativo Enzimas antioxidantes Drosophila melanogaster Oxidative stress Antioxidants enzymes CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA |
topic |
Caryocar coriaceum Wittm. Pequi Estresse oxidativo Enzimas antioxidantes Drosophila melanogaster Oxidative stress Antioxidants enzymes CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA |
description |
Despite the diversity of species found in the National Araripe Forest, Ceará Cariri, there are few studies with Caryocar coriaceumWittm (C. coriaceum), a plant that produces a highly consumed fruit in regional cuisine. The oil extracted from the pulp is also widely used in popular medicine for the treatment of respiratory infections and joint diseases. Studies with oil pulp have consolidated therapeutic properties of species. However, this is the first study with plant leaf extracts and oil from the fruit pulp that assesses markers of oxidative stress and gene expression of antioxidant enzymes in vivo using Drosophila melanogaster. The antioxidant activity of leaf and fruit peel extracts was evaluated in vitro. The major phenolic compounds from extracts, identified by HPLC-DAD, were quercetin, chlorogenic acid and ellagic. In in vitro tests, the extracts exhibited similar and significant DPPH● and OH● scavenging activity, were effective in reducing and chelating iron as well as decreasing lipid peroxidation and production of reactive oxygen species induced by iron and calcium, respectively, in liver homogenate. The extracts did not change the levels O2•– in mitochondria treated with antimycin A. In in vivo study, flies were pretreated with a diet containing aqueous leaf extract (AEL) (1 and 5 mg/g diet) and/or pequi pulp oil (PPO) (1, 5 and 10mg/g diet) for 7 days followed by exposure to paraquat (PQ) (10mM) for 24 hours. A third treatment was performed concomitantly with AEL and PQ (1mM) for 5 days. The analysis of the negative geotaxia was performed to evaluate locomotor ability of flies. Flies homogenate was used to analyze oxidative stress markers specifically activity of catalase (CAT) and glutathione-s-transferase (GST), levels of ROS and peroxidation lipid by levels of TBARS. We evaluated the mRNA expression of antioxidant enzyme genes CAT, superoxide dismutase (SOD), Thioredoxin reductase-1 (Trx R-1), Thioredoxin T (Trx T) and Keap 1 protein by qRT-PCR. Flies exposed to PQ showed a significant impairment in locomotor activity, increase in oxidative stress markers and gene expression of CAT, SOD, Trx R-1, Trx T and Keap 1. The concentrations AEL and OPP, in all treatments, reduced significantly lipid peroxidation levels and reversed locomotor changes induced by exposure to PQ. In both AEL treatments, flies exposed to PQ had a reduction in ROS levels and CAT and GST activities. Flies concomitantly treated with AEL and PQ exhibited a significant decrease in expression of Trx R-1 and Trx T genes, which were increased in the PQ group. Pre-treatment with PPO of 10mg/g diet, we verified a decrease the expression of genes CAT, SOD, Keap 1 and Trx T, which were increased in the group exposed to PQ alone. The data obtained in this study show that the treatment with AEL and PPO reduced oxidative stress markers induced by PQ and suggest that this effect may be related, at least in part, to the antioxidant system modulation. Moreover, these results will collaborate to extend the knowledge about the scientific properties of C. coriaceum as well as point it as a promissing functional food. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-04-29 2019-08-19T19:45:45Z 2019-08-19T19:45:45Z |
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/1/17965 |
url |
http://repositorio.ufsm.br/handle/1/17965 |
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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1814349575794720768 |