Avaliação do caráter oxidante da violaceína

Detalhes bibliográficos
Ano de defesa: 2011
Autor(a) principal: Leal, Angélica Maria de Sousa
Orientador(a): Não Informado pela instituição
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal do Rio Grande do Norte
BR
UFRN
Programa de Pós-Graduação em Bioquímica
Bioquímica; Biologia Molecular
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: https://repositorio.ufrn.br/jspui/handle/123456789/18533
Resumo: Violacein is a violet pigment isolated from many gram-negative bacteria, especially from Chromobacterium violaceum, a betaproteobacterium found in the Amazon River in Brazil. It has potential medical applications as an antibacterial, fungicide, anti-tryptanocidal, anti-ulcerogenic and anti-cancer drug, among others. Furthermore, its pro-oxidant activity has been suggested, but only in two specific tumor lineages. Thus, in the present study, the prooxidant effects of violacein were investigated in both normal and tumor cells, seeking to evaluate the cell responses. The evaluation of violacein cytotoxicity using the Trypan blue dye exclusion method indicated that CHO-K1 cells were more resistant than tumor HeLa cells. The oxidative stress induced by violacein was manifested as an increase in intracellular SOD activity in CHO-K1 and MRC-5 cells at a specific concentration range. Nevertheless, a decrease was detected specifically at 6-12 μM in HeLa and MRC-5 cells. Interestingly, the increase in SOD activity was not followed by a concomitant increase in catalase activity. Regarding to oxidative stress biomarkers, increased protein carbonylation and lipid hydroperoxides levels were detected respectively in CHO-K1 and MRC-5 cells treated with violacein at 1.5-3 μM and 3 μM, which may be an evidence that this compound causes oxidative stress specifically in these conditions. Additionally, it is believed that the decline in cell viability observed in MRC-5 cells and HeLa treated with violacein at 6-12 M is due to mechanisms not related to oxidative stress. Moreover, the results suggested that violacein might cause oxidative stress by increasing endogenous levels of O2 -, since the occurrence of an expressive change in SOD activity. In addition, in order to evaluate the antioxidant activity of violacein in the absence of a biological system, the total antioxidant and iron chelating activity were evaluated, so that antioxidant activities were detected at 30 and 60 μM of violacein. Altogether, the results indicate that although oxidative stress is triggered by incubation with violacein, it did not seem to be high enough to cause serious damage to cell biomolecules in HeLa cells and only at specific concentrations in CHOK-1 and MRC-5 cells. Comparing the results obtained in cell culture and the in vitro antioxidant activity evaluation, the results confirmed that violacein presents opposing oxidant features when in presence or absence of a biological system and the antioxidant character only occurs at high concentrations of the pigment.
id UFRN_72c88da503e2966f5e95f186a809a797
oai_identifier_str oai:repositorio.ufrn.br:123456789/18533
network_acronym_str UFRN
network_name_str Repositório Institucional da UFRN
repository_id_str
spelling Avaliação do caráter oxidante da violaceínaAvaliação do caráter oxidante da violaceínaViolaceínaPotencial oxidante da violaceínaAtividade das enzimas do aparato antioxidanteBiomarcadores de estresse oxidativo.ViolaceinOxidant potential of violaceinActivity of enzymatic antioxidant apparatusBiomarkers of oxidative stress.CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICAViolacein is a violet pigment isolated from many gram-negative bacteria, especially from Chromobacterium violaceum, a betaproteobacterium found in the Amazon River in Brazil. It has potential medical applications as an antibacterial, fungicide, anti-tryptanocidal, anti-ulcerogenic and anti-cancer drug, among others. Furthermore, its pro-oxidant activity has been suggested, but only in two specific tumor lineages. Thus, in the present study, the prooxidant effects of violacein were investigated in both normal and tumor cells, seeking to evaluate the cell responses. The evaluation of violacein cytotoxicity using the Trypan blue dye exclusion method indicated that CHO-K1 cells were more resistant than tumor HeLa cells. The oxidative stress induced by violacein was manifested as an increase in intracellular SOD activity in CHO-K1 and MRC-5 cells at a specific concentration range. Nevertheless, a decrease was detected specifically at 6-12 μM in HeLa and MRC-5 cells. Interestingly, the increase in SOD activity was not followed by a concomitant increase in catalase activity. Regarding to oxidative stress biomarkers, increased protein carbonylation and lipid hydroperoxides levels were detected respectively in CHO-K1 and MRC-5 cells treated with violacein at 1.5-3 μM and 3 μM, which may be an evidence that this compound causes oxidative stress specifically in these conditions. Additionally, it is believed that the decline in cell viability observed in MRC-5 cells and HeLa treated with violacein at 6-12 M is due to mechanisms not related to oxidative stress. Moreover, the results suggested that violacein might cause oxidative stress by increasing endogenous levels of O2 -, since the occurrence of an expressive change in SOD activity. In addition, in order to evaluate the antioxidant activity of violacein in the absence of a biological system, the total antioxidant and iron chelating activity were evaluated, so that antioxidant activities were detected at 30 and 60 μM of violacein. Altogether, the results indicate that although oxidative stress is triggered by incubation with violacein, it did not seem to be high enough to cause serious damage to cell biomolecules in HeLa cells and only at specific concentrations in CHOK-1 and MRC-5 cells. Comparing the results obtained in cell culture and the in vitro antioxidant activity evaluation, the results confirmed that violacein presents opposing oxidant features when in presence or absence of a biological system and the antioxidant character only occurs at high concentrations of the pigment.Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorA violaceína é um pigmento violeta isolado de várias espécies de bactérias gram-negativas, especialmente da Chromobacterium violaceum, uma betaproteobactéria encontrada no rio Amazonas, no Brasil. Diversas atividades biológicas já foram descritas para este pigmento e dentre elas destacam-se a antibacteriana, antifúngica, tripanocida, antileishmaniose, anti-úlcerogênica, antiviral e antitumoral. Apesar de uma atividade antioxidante in vitro ter sido sugerida, a atividade pró-oxidante também já foi observada especificamente em duas linhagens tumorais e parece ser dependente de mecanismos específicos para cada linhagem. Nesse sentido, os efeitos citotóxicos e próoxidantes da violaceína foram investigados em células normais e tumorais buscando-se avaliar a ocorrência de diferentes respostas celulares. A análise da citotoxicidade da violaceína indicou que células CHO-K1 foram mais resistentes ao composto em relação às tumorais HeLa. Quanto às enzimas do aparato antioxidante, observou-se um aumento significativo na atividade da SOD intracelular nas linhagens CHO-K1 e MRC-5. Porém, houve uma diminuição na atividade enzimática especificamente nos tratamentos com 6 e 12 M nas linhagens MRC-5 e HeLa. Interessantemente, o aumento na atividade da SOD não foi acompanhado pelo aumento concomitante na atividade da catalase. Em relação aos biomarcadores de estresse oxidativo, níveis elevados de proteínas carboniladas e hidroperóxidos de lipídio foram observados em células CHO-K1 e MRC-5 quando tratadas respectivamente com 1,5-3 μM e 3 μM de violaceína, indicando que o pigmento apresenta efeitos pró-oxidantes especificamente nessas concentrações. Adicionalmente, acredita-se que a acentuada queda na viabilidade celular observada em células MRC-5 e HeLa tratadas com 6-12 μM de violaceína se deve a outros mecanismos não relacionados à geração de estresse oxidativo propriamente dita. Os resultados obtidos em cultura de células sugerem também que a violaceína induz estresse oxidativo por elevação dos níveis endógenos de O2 •–, visto a ocorrência de uma significativa alteração nos níveis de atividade de SOD. Em adição, com o objetivo de avaliar o caráter antioxidante in vitro da violaceína na ausência de um sistema biológico celular, a capacidade antioxidante total e a atividade de quelação férrica do pigmento foram avaliadas, de forma que atividades antioxidantes foram detectadas a 30 e 60 μM de violaceína. Frente aos resultados obtidos, apesar do desencadeamento do estresse oxidativo após a incubação com violaceína, este parece não ser suficiente para causar danos significativos aos componentes e estruturas celulares em células HeLa e apenas em concentrações específicas de pigmento para CHO-K1 e MRC-5, dentro das condições avaliadas. Por fim, os resultados confirmam que violaceína apresenta caráteres oxidantes opostos quando na presença ou ausência de um sistema biológico, além de que o caráter antioxidante só se dá em concentrações elevadas do pigmentoUniversidade Federal do Rio Grande do NorteBRUFRNPrograma de Pós-Graduação em BioquímicaBioquímica; Biologia Molecularhttp://lattes.cnpq.br/2028668396962984http://lattes.cnpq.br/1083882171718362Bonatto, Diegohttp://lattes.cnpq.br/2915446717051640Vieira, Valéria Karla de Britohttp://lattes.cnpq.br/2840407921428106Leal, Angélica Maria de Sousa2015-03-03T13:58:19Z2015-02-252015-03-03T13:58:19Z2011-09-28info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfapplication/pdfLEAL, Angélica Maria de Sousa. Avaliação do caráter oxidante da violaceína. 2011. 81 f. Dissertação (Mestrado em Bioquímica; Biologia Molecular) - Universidade Federal do Rio Grande do Norte, Natal, 2011.https://repositorio.ufrn.br/jspui/handle/123456789/18533porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFRNinstname:Universidade Federal do Rio Grande do Norte (UFRN)instacron:UFRN2019-01-30T01:33:43Zoai:repositorio.ufrn.br:123456789/18533Repositório InstitucionalPUBhttp://repositorio.ufrn.br/oai/repositorio@bczm.ufrn.bropendoar:2019-01-30T01:33:43Repositório Institucional da UFRN - Universidade Federal do Rio Grande do Norte (UFRN)false
dc.title.none.fl_str_mv Avaliação do caráter oxidante da violaceína
Avaliação do caráter oxidante da violaceína
title Avaliação do caráter oxidante da violaceína
spellingShingle Avaliação do caráter oxidante da violaceína
Leal, Angélica Maria de Sousa
Violaceína
Potencial oxidante da violaceína
Atividade das enzimas do aparato antioxidante
Biomarcadores de estresse oxidativo.
Violacein
Oxidant potential of violacein
Activity of enzymatic antioxidant apparatus
Biomarkers of oxidative stress.
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
title_short Avaliação do caráter oxidante da violaceína
title_full Avaliação do caráter oxidante da violaceína
title_fullStr Avaliação do caráter oxidante da violaceína
title_full_unstemmed Avaliação do caráter oxidante da violaceína
title_sort Avaliação do caráter oxidante da violaceína
author Leal, Angélica Maria de Sousa
author_facet Leal, Angélica Maria de Sousa
author_role author
dc.contributor.none.fl_str_mv
http://lattes.cnpq.br/2028668396962984

http://lattes.cnpq.br/1083882171718362
Bonatto, Diego

http://lattes.cnpq.br/2915446717051640
Vieira, Valéria Karla de Brito

http://lattes.cnpq.br/2840407921428106
dc.contributor.author.fl_str_mv Leal, Angélica Maria de Sousa
dc.subject.por.fl_str_mv Violaceína
Potencial oxidante da violaceína
Atividade das enzimas do aparato antioxidante
Biomarcadores de estresse oxidativo.
Violacein
Oxidant potential of violacein
Activity of enzymatic antioxidant apparatus
Biomarkers of oxidative stress.
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
topic Violaceína
Potencial oxidante da violaceína
Atividade das enzimas do aparato antioxidante
Biomarcadores de estresse oxidativo.
Violacein
Oxidant potential of violacein
Activity of enzymatic antioxidant apparatus
Biomarkers of oxidative stress.
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
description Violacein is a violet pigment isolated from many gram-negative bacteria, especially from Chromobacterium violaceum, a betaproteobacterium found in the Amazon River in Brazil. It has potential medical applications as an antibacterial, fungicide, anti-tryptanocidal, anti-ulcerogenic and anti-cancer drug, among others. Furthermore, its pro-oxidant activity has been suggested, but only in two specific tumor lineages. Thus, in the present study, the prooxidant effects of violacein were investigated in both normal and tumor cells, seeking to evaluate the cell responses. The evaluation of violacein cytotoxicity using the Trypan blue dye exclusion method indicated that CHO-K1 cells were more resistant than tumor HeLa cells. The oxidative stress induced by violacein was manifested as an increase in intracellular SOD activity in CHO-K1 and MRC-5 cells at a specific concentration range. Nevertheless, a decrease was detected specifically at 6-12 μM in HeLa and MRC-5 cells. Interestingly, the increase in SOD activity was not followed by a concomitant increase in catalase activity. Regarding to oxidative stress biomarkers, increased protein carbonylation and lipid hydroperoxides levels were detected respectively in CHO-K1 and MRC-5 cells treated with violacein at 1.5-3 μM and 3 μM, which may be an evidence that this compound causes oxidative stress specifically in these conditions. Additionally, it is believed that the decline in cell viability observed in MRC-5 cells and HeLa treated with violacein at 6-12 M is due to mechanisms not related to oxidative stress. Moreover, the results suggested that violacein might cause oxidative stress by increasing endogenous levels of O2 -, since the occurrence of an expressive change in SOD activity. In addition, in order to evaluate the antioxidant activity of violacein in the absence of a biological system, the total antioxidant and iron chelating activity were evaluated, so that antioxidant activities were detected at 30 and 60 μM of violacein. Altogether, the results indicate that although oxidative stress is triggered by incubation with violacein, it did not seem to be high enough to cause serious damage to cell biomolecules in HeLa cells and only at specific concentrations in CHOK-1 and MRC-5 cells. Comparing the results obtained in cell culture and the in vitro antioxidant activity evaluation, the results confirmed that violacein presents opposing oxidant features when in presence or absence of a biological system and the antioxidant character only occurs at high concentrations of the pigment.
publishDate 2011
dc.date.none.fl_str_mv 2011-09-28
2015-03-03T13:58:19Z
2015-02-25
2015-03-03T13:58:19Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv LEAL, Angélica Maria de Sousa. Avaliação do caráter oxidante da violaceína. 2011. 81 f. Dissertação (Mestrado em Bioquímica; Biologia Molecular) - Universidade Federal do Rio Grande do Norte, Natal, 2011.
https://repositorio.ufrn.br/jspui/handle/123456789/18533
identifier_str_mv LEAL, Angélica Maria de Sousa. Avaliação do caráter oxidante da violaceína. 2011. 81 f. Dissertação (Mestrado em Bioquímica; Biologia Molecular) - Universidade Federal do Rio Grande do Norte, Natal, 2011.
url https://repositorio.ufrn.br/jspui/handle/123456789/18533
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Universidade Federal do Rio Grande do Norte
BR
UFRN
Programa de Pós-Graduação em Bioquímica
Bioquímica; Biologia Molecular
publisher.none.fl_str_mv Universidade Federal do Rio Grande do Norte
BR
UFRN
Programa de Pós-Graduação em Bioquímica
Bioquímica; Biologia Molecular
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFRN
instname:Universidade Federal do Rio Grande do Norte (UFRN)
instacron:UFRN
instname_str Universidade Federal do Rio Grande do Norte (UFRN)
instacron_str UFRN
institution UFRN
reponame_str Repositório Institucional da UFRN
collection Repositório Institucional da UFRN
repository.name.fl_str_mv Repositório Institucional da UFRN - Universidade Federal do Rio Grande do Norte (UFRN)
repository.mail.fl_str_mv repositorio@bczm.ufrn.br
_version_ 1855758722461073408