Caracterização físico-química e atividade da Curcuma longa L. em células endoteliais sob estresse oxidativo
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFG |
dARK ID: | ark:/38995/0013000001f9s |
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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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/10424ark:/38995/0013000001f9sIschemic 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). 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) Previous issue date: 2020-02-28application/pdfporUniversidade Federal de GoiásPrograma de Pós-graduação em Ciência Animal (EVZ)UFGBrasilEscola de Veterinária e Zootecnia - EVZ (RG)http://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessÁcido gálicoCurcuminaDPPHEstresse oxidativoHUVECGallic acidCurcuminOxidative stressCIENCIAS AGRARIAS::MEDICINA VETERINARIACaracterização físico-química e atividade da Curcuma longa L. em células endoteliais sob estresse oxidativoPhysical-chemical analysis and activity of Curcuma longa L. in endothelial cells under oxidative stressinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis4581960685150189167600600600-6217552114249094582453670264235017319reponame:Repositório Institucional da UFGinstname:Universidade Federal de Goiás (UFG)instacron:UFGLICENSElicense.txtlicense.txttext/plain; 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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 |
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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. ark:/38995/0013000001f9s |
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http://repositorio.bc.ufg.br/tede/handle/tede/10424 |
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por |
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por |
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openAccess |
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Universidade Federal de Goiás |
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Programa de Pós-graduação em Ciência Animal (EVZ) |
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UFG |
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Brasil |
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Escola de Veterinária e Zootecnia - EVZ (RG) |
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Universidade Federal de Goiás |
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