Estudo do potencial antioxidante da espécie Caryocar coriaceum Wittm. em modelos experimentais in vitro e in vivo

Detalhes bibliográficos
Ano de defesa: 2016
Autor(a) principal: Pereira, Sandra Mara Pimentel Duavy
Orientador(a): Não Informado pela instituição
Banca de defesa: Não Informado pela instituição
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.
id UFSM_16738dc5515204a9cb03ad8787ea663d
oai_identifier_str oai:repositorio.ufsm.br:1/17965
network_acronym_str UFSM
network_name_str Manancial - Repositório Digital da UFSM
repository_id_str
spelling 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
_version_ 1814349575794720768