Caracterização físico-química e atividade da Curcuma longa L. em células endoteliais sob estresse oxidativo

Detalhes bibliográficos
Autor(a) principal: Teles, Juliana Carvalho de Almeida Borges
Data de Publicação: 2020
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFG
Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/10424
Resumo: Ischemic tissue injury is quite harmful, however, reperfusion causes an exacerbated level of reactive oxygen species (ROS), which directly interfere in the cell signaling cascade, with cell deleterious effects such as lipid peroxidation, protein oxidation and DNA damage. Researches had shown the beneficial effects of antioxidants present in vegetables and among them is curcumin, an antioxidant pigment found in Curcuma longa L. rhizomes, known as turmeric or saffron. This plant has demonstrated pharmacological activities, which have been reported since antiquity, as antioxidant, anti-inflammatory, antibacterial, antifungal, antitumor, antiparasitic, antiviral, healing, hypoglycemic, neuroprotective and immunomodulatory. Therefore, the aim of this research was to evaluate the possible antioxidant action of curcumin on human umbilical vein endothelial cells (HUVEC), in culture, submitted and not submitted to oxidative stress (EO). This study was divided into three phases. In the first time, the action of standard curcumin (Sigma Aldrich, Reference C1386) was evaluated at different concentrations (0 μmol, 20 μmol, 50 μmol, 100 μmol and 1000 μmol) and for three different times of continuous exposure (24 h, 48 h h and 72 h). The EO assay was induced through cellular exposure to 1% hydrogen peroxide and through the MTT (methyl-thiazolyl-tetrazolium) reduction assay, cell viability and cytotoxicity could be assessed. In the second phase, we determinated the presence and content of antioxidant compounds in the ethanol extract of Curcuma longa L. (EEC) from Mara Rosa-GO. EEC samples were subjected to Folin-Ciocalteau tests, capture of the DPPH radical (2,2-diphenyl-1-picryl-hydrazil) and the HPLC-Fluorescence method. In the third phase of this research, the effect of EEC in HUVEC cultures was evaluated under the same experimental conditions as the first phase, in addition with the possible signaling pathways of both survival and cell death. The results showed that the cells treated with 20 μmol of standard curcumin and at 24 h and 48 h showed better cell viability and, consequently, less cytotoxicity. The occurrence of oxidative stress was validated by the decrease in cell viability within the three times variables used, but an antioxidative protection was found at 50 μmol of curcumin. In EEC, we identified gallic acid, curcumin (16.7%) and the amount of 18.1 μg / mL of extract, to reduce the DPPH radical by 50%. Results from the third phase indicated that the EEC possibly alleviated cell damage by oxidative stress at the dosage of 100 μg / ml, by decreasing the immunoexpression of caspase3 and by increasing the signaling of the SIRT1 survival route. It was concluded that both pure curcumin and EEC can improve the cell viability of HUVEC under oxidative stress.
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spelling Araújo , Eugênio Gonçalves dehttp://lattes.cnpq.br/3919777570059928Saturnino, Klaus Casarohttp://lattes.cnpq.br/6894735942112278Cruz, Vanessa de Sousahttp://lattes.cnpq.br/8788967925940484Araújo, Eugênio Gonçalves deIglesias, Ana PaulaDamasceno, Adilson DonizeteBeretta, Daniel CortêsArantes , Diego Antônio Costahttp://lattes.cnpq.br/6946398230607133Teles, Juliana Carvalho de Almeida Borges2020-03-16T13:46:23Z2020-02-28TELES, Juliana Carvalho de Almeida Borges. Caracterização físico-química e atividade da Curcuma longa L. em células endoteliais sob estresse oxidativo. 2020. 64 f. Tese (Doutorado em Ciência Animal) - Universidade Federal de Goiás, Goiânia, 2020.http://repositorio.bc.ufg.br/tede/handle/tede/10424Ischemic tissue injury is quite harmful, however, reperfusion causes an exacerbated level of reactive oxygen species (ROS), which directly interfere in the cell signaling cascade, with cell deleterious effects such as lipid peroxidation, protein oxidation and DNA damage. Researches had shown the beneficial effects of antioxidants present in vegetables and among them is curcumin, an antioxidant pigment found in Curcuma longa L. rhizomes, known as turmeric or saffron. This plant has demonstrated pharmacological activities, which have been reported since antiquity, as antioxidant, anti-inflammatory, antibacterial, antifungal, antitumor, antiparasitic, antiviral, healing, hypoglycemic, neuroprotective and immunomodulatory. Therefore, the aim of this research was to evaluate the possible antioxidant action of curcumin on human umbilical vein endothelial cells (HUVEC), in culture, submitted and not submitted to oxidative stress (EO). This study was divided into three phases. In the first time, the action of standard curcumin (Sigma Aldrich, Reference C1386) was evaluated at different concentrations (0 μmol, 20 μmol, 50 μmol, 100 μmol and 1000 μmol) and for three different times of continuous exposure (24 h, 48 h h and 72 h). The EO assay was induced through cellular exposure to 1% hydrogen peroxide and through the MTT (methyl-thiazolyl-tetrazolium) reduction assay, cell viability and cytotoxicity could be assessed. In the second phase, we determinated the presence and content of antioxidant compounds in the ethanol extract of Curcuma longa L. (EEC) from Mara Rosa-GO. EEC samples were subjected to Folin-Ciocalteau tests, capture of the DPPH radical (2,2-diphenyl-1-picryl-hydrazil) and the HPLC-Fluorescence method. In the third phase of this research, the effect of EEC in HUVEC cultures was evaluated under the same experimental conditions as the first phase, in addition with the possible signaling pathways of both survival and cell death. The results showed that the cells treated with 20 μmol of standard curcumin and at 24 h and 48 h showed better cell viability and, consequently, less cytotoxicity. The occurrence of oxidative stress was validated by the decrease in cell viability within the three times variables used, but an antioxidative protection was found at 50 μmol of curcumin. In EEC, we identified gallic acid, curcumin (16.7%) and the amount of 18.1 μg / mL of extract, to reduce the DPPH radical by 50%. Results from the third phase indicated that the EEC possibly alleviated cell damage by oxidative stress at the dosage of 100 μg / ml, by decreasing the immunoexpression of caspase3 and by increasing the signaling of the SIRT1 survival route. It was concluded that both pure curcumin and EEC can improve the cell viability of HUVEC under oxidative stress.A injúria tecidual por isquemia é bastante nociva e, em situações de reperfusão, pode provocar a formação exacerbada de espécies reativas do oxigênio (ERO), que interferem diretamente na cascata de sinalização celular, com efeitos deletérios como a peroxidação lipídica da membrana celular, a oxidação de proteínas e a lesão ao DNA. As pesquisas têm mostrado os efeitos benéficos dos antioxidantes presentes nos vegetais e, entre eles encontra-se a curcumina, um pigmento antioxidante do rizoma da Curcuma longa L., conhecida como açafrão ou cúrcuma. Essa planta demonstra atividades farmacológicas, que são relatadas desde a antiguidade, como antioxidante, anti-inflamatório, antibacteriano, antifúngico, antitumoral, antiparasitário, antiviral, cicatrizante, hipoglicemiante, neuroprotetor e imunomodulador. Diante disso, objetivou-se com essa pesquisa avaliar a possível ação antioxidante da curcumina sobre as células endoteliais de veia umbilical humana (HUVEC), em cultivo, submetidas e não submetidas ao estresse oxidativo (EO). Este estudo foi dividido em três fases. Na primeira fase avaliou-se a ação da curcumina padrão (Sigma Aldrich, Referência C1386) em diferentes concentrações (0 μmol, 20 μmol, 50 μmol, 100 μmol e 1000 μmol) e por três tempos diferentes de exposição contínua (24 h, 48 h e 72 h). O ensaio de EO foi induzido por meio da exposição celular ao peróxido de hidrogênio a 1% e por meio do ensaio de redução do MTT (metil-tiazolil-tetrazólio), pôde-se avaliar a viabilidade celular e a citotoxicidade. Na segunda fase do estudo determinou-se a presença e o teor de compostos antioxidantes no extrato etanólico da Curcuma longa L. (EEC) proveniente de Mara Rosa-GO. As amostras do EEC foram submetidas aos testes de Folin-Ciocalteau, à captura do radical DPPH (2,2-difenil-1- picril-hidrazil) e ao método de HPLC-Fluorescence. Na terceira fase da pesquisa avaliou-se o efeito do EEC em cultivos de HUVEC sob as mesmas condições experimentais da primeira fase, além disso, pesquisou-se as vias de sinalização tanto de sobrevivência, quanto de morte celular possivelmente ocorridas. Os resultados demonstraram que as células tratadas com 20 μmol de curcumina padrão e nos tempos de 24 h e 48 h apresentaram melhor viabilidade celular e consequentemente, menor citotoxicidade. A ocorrência do estresse oxidativo foi validada pelo decréscimo na viabilidade celular dentro das três variáveis de tempo utilizadas, com uma proteção antioxidativa na dosagem de 50 μmol de curcumina. No EEC pôde-se identificar o ácido gálico, a curcumina (16,7%) e a quantidade de 18,1 μg/mL de extrato, para se reduzir em 50% o radical DPPH. Os resultados da terceira fase indicaram que o EEC possivelmente amenizou a injúria celular por estresse oxidativo, na dosagem de 100 μg/ml, por diminuição da imunoexpressão de caspase3 e pelo aumento da sinalização da rota de sobrevivência SIRT1. Concluiu-se que tanto a curcumina pura, quanto o EEC pode aumentar a viabilidade celular de células endoteliais sob estresse oxidativo.Submitted by Liliane Ferreira (ljuvencia30@gmail.com) on 2020-03-13T13:04:19Z No. of bitstreams: 2 Tese - Juliana Carvalho de Almeida Borges Teles - 2020.pdf: 5270611 bytes, checksum: a9c07cc022f1ada8d5c267a863421667 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2020-03-16T13:46:23Z (GMT) No. of bitstreams: 2 Tese - Juliana Carvalho de Almeida Borges Teles - 2020.pdf: 5270611 bytes, checksum: a9c07cc022f1ada8d5c267a863421667 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2020-03-16T13:46:23Z (GMT). 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dc.title.eng.fl_str_mv Caracterização físico-química e atividade da Curcuma longa L. em células endoteliais sob estresse oxidativo
dc.title.alternative.eng.fl_str_mv Physical-chemical analysis and activity of Curcuma longa L. in endothelial cells under oxidative stress
title Caracterização físico-química e atividade da Curcuma longa L. em células endoteliais sob estresse oxidativo
spellingShingle Caracterização físico-química e atividade da Curcuma longa L. em células endoteliais sob estresse oxidativo
Teles, Juliana Carvalho de Almeida Borges
Ácido gálico
Curcumina
DPPH
Estresse oxidativo
HUVEC
Gallic acid
Curcumin
Oxidative stress
CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Caracterização físico-química e atividade da Curcuma longa L. em células endoteliais sob estresse oxidativo
title_full Caracterização físico-química e atividade da Curcuma longa L. em células endoteliais sob estresse oxidativo
title_fullStr Caracterização físico-química e atividade da Curcuma longa L. em células endoteliais sob estresse oxidativo
title_full_unstemmed Caracterização físico-química e atividade da Curcuma longa L. em células endoteliais sob estresse oxidativo
title_sort Caracterização físico-química e atividade da Curcuma longa L. em células endoteliais sob estresse oxidativo
author Teles, Juliana Carvalho de Almeida Borges
author_facet Teles, Juliana Carvalho de Almeida Borges
author_role author
dc.contributor.advisor1.fl_str_mv Araújo , Eugênio Gonçalves de
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/3919777570059928
dc.contributor.advisor-co1.fl_str_mv Saturnino, Klaus Casaro
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/6894735942112278
dc.contributor.advisor-co2.fl_str_mv Cruz, Vanessa de Sousa
dc.contributor.advisor-co2Lattes.fl_str_mv http://lattes.cnpq.br/8788967925940484
dc.contributor.referee1.fl_str_mv Araújo, Eugênio Gonçalves de
dc.contributor.referee2.fl_str_mv Iglesias, Ana Paula
dc.contributor.referee3.fl_str_mv Damasceno, Adilson Donizete
dc.contributor.referee4.fl_str_mv Beretta, Daniel Cortês
dc.contributor.referee5.fl_str_mv Arantes , Diego Antônio Costa
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/6946398230607133
dc.contributor.author.fl_str_mv Teles, Juliana Carvalho de Almeida Borges
contributor_str_mv Araújo , Eugênio Gonçalves de
Saturnino, Klaus Casaro
Cruz, Vanessa de Sousa
Araújo, Eugênio Gonçalves de
Iglesias, Ana Paula
Damasceno, Adilson Donizete
Beretta, Daniel Cortês
Arantes , Diego Antônio Costa
dc.subject.por.fl_str_mv Ácido gálico
Curcumina
DPPH
Estresse oxidativo
HUVEC
topic Ácido gálico
Curcumina
DPPH
Estresse oxidativo
HUVEC
Gallic acid
Curcumin
Oxidative stress
CIENCIAS AGRARIAS::MEDICINA VETERINARIA
dc.subject.eng.fl_str_mv Gallic acid
Curcumin
Oxidative stress
dc.subject.cnpq.fl_str_mv CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description Ischemic tissue injury is quite harmful, however, reperfusion causes an exacerbated level of reactive oxygen species (ROS), which directly interfere in the cell signaling cascade, with cell deleterious effects such as lipid peroxidation, protein oxidation and DNA damage. Researches had shown the beneficial effects of antioxidants present in vegetables and among them is curcumin, an antioxidant pigment found in Curcuma longa L. rhizomes, known as turmeric or saffron. This plant has demonstrated pharmacological activities, which have been reported since antiquity, as antioxidant, anti-inflammatory, antibacterial, antifungal, antitumor, antiparasitic, antiviral, healing, hypoglycemic, neuroprotective and immunomodulatory. Therefore, the aim of this research was to evaluate the possible antioxidant action of curcumin on human umbilical vein endothelial cells (HUVEC), in culture, submitted and not submitted to oxidative stress (EO). This study was divided into three phases. In the first time, the action of standard curcumin (Sigma Aldrich, Reference C1386) was evaluated at different concentrations (0 μmol, 20 μmol, 50 μmol, 100 μmol and 1000 μmol) and for three different times of continuous exposure (24 h, 48 h h and 72 h). The EO assay was induced through cellular exposure to 1% hydrogen peroxide and through the MTT (methyl-thiazolyl-tetrazolium) reduction assay, cell viability and cytotoxicity could be assessed. In the second phase, we determinated the presence and content of antioxidant compounds in the ethanol extract of Curcuma longa L. (EEC) from Mara Rosa-GO. EEC samples were subjected to Folin-Ciocalteau tests, capture of the DPPH radical (2,2-diphenyl-1-picryl-hydrazil) and the HPLC-Fluorescence method. In the third phase of this research, the effect of EEC in HUVEC cultures was evaluated under the same experimental conditions as the first phase, in addition with the possible signaling pathways of both survival and cell death. The results showed that the cells treated with 20 μmol of standard curcumin and at 24 h and 48 h showed better cell viability and, consequently, less cytotoxicity. The occurrence of oxidative stress was validated by the decrease in cell viability within the three times variables used, but an antioxidative protection was found at 50 μmol of curcumin. In EEC, we identified gallic acid, curcumin (16.7%) and the amount of 18.1 μg / mL of extract, to reduce the DPPH radical by 50%. Results from the third phase indicated that the EEC possibly alleviated cell damage by oxidative stress at the dosage of 100 μg / ml, by decreasing the immunoexpression of caspase3 and by increasing the signaling of the SIRT1 survival route. It was concluded that both pure curcumin and EEC can improve the cell viability of HUVEC under oxidative stress.
publishDate 2020
dc.date.accessioned.fl_str_mv 2020-03-16T13:46:23Z
dc.date.issued.fl_str_mv 2020-02-28
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.citation.fl_str_mv TELES, Juliana Carvalho de Almeida Borges. Caracterização físico-química e atividade da Curcuma longa L. em células endoteliais sob estresse oxidativo. 2020. 64 f. Tese (Doutorado em Ciência Animal) - Universidade Federal de Goiás, Goiânia, 2020.
dc.identifier.uri.fl_str_mv http://repositorio.bc.ufg.br/tede/handle/tede/10424
identifier_str_mv TELES, Juliana Carvalho de Almeida Borges. Caracterização físico-química e atividade da Curcuma longa L. em células endoteliais sob estresse oxidativo. 2020. 64 f. Tese (Doutorado em Ciência Animal) - Universidade Federal de Goiás, Goiânia, 2020.
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dc.publisher.none.fl_str_mv Universidade Federal de Goiás
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dc.publisher.department.fl_str_mv Escola de Veterinária e Zootecnia - EVZ (RG)
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