Efeito cito-genômico do peróxido de hidrogênio e do guaraná (Paullinia cupana) em células tronco mesenquimais
Ano de defesa: | 2014 |
---|---|
Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | , |
Tipo de documento: | Dissertação |
Tipo de acesso: | Acesso aberto |
Idioma: | por |
Instituição de defesa: |
Universidade Federal de Santa Maria
|
Programa de Pós-Graduação: |
Programa de Pós-Graduação em Farmacologia
|
Departamento: |
Farmacologia
|
País: |
BR
|
Palavras-chave em Português: | |
Palavras-chave em Inglês: | |
Área do conhecimento CNPq: | |
Link de acesso: | http://repositorio.ufsm.br/handle/1/8989 |
Resumo: | With advances in techniques of cell culture, stem cells (SCs) have to be used as an alternative therapy, firming the called regenerative medicine. An type of SC that has been studied is the Adipose-derived stem cell (ASCs) having identical characteristics of Mesenchymal Stem Cells (MSCs). To realize a transplanting of SCs, is necessary a large amount of cells, which makes such cultures are maintained in vitro for long periods of time, consequently these cells, with the repeated passages, enter cellular senescence, which prevents their use. Thus, some studies showed that the hydrogen peroxide (H2O2) can act to potentiate the proliferation of these cells. However, it is know that the oxidative metabolism can be associated with the senescence process. From this, the aim of this study was available the cito-genomic effect of the exposition of ASCs to H2O2 and to guaraná extract. Was realized isolation and cultivation of ASCs, that were, in forth passage, exposed to concentrations of 1-1000 μM of H2O2 and realized assays to available the cell viability, oxidative stress parameters, DNA damage and the apoptotic way of caspases. Moreover, senecents ASCs were treated with concentrations of 1-20 mg/mL of guaraná extract, being tested to the senescence reversion, cell viability, oxidative parameters, DNA damage and activity and expression of antioxidants enzymes. The results showed that the H2O2 cause cito-genomic damages to ASCs, with decrease in the cell viability mainly at the concentration of 200 μM, as well as increase the total reactive oxygen species (ROS) rate and lipidic peroxidation effect. The H2O2 also decreased the catalase activity in dose-dependent form e acted in the SOD activity. The treatments also conducted to a progressive increase of DNA damage. Moreover, in the eighth passage was observed the senescence of ASCs and the treatment with guaraná showed capacity to revert this phenotype in the concentration of 5 mg/mL, with reduction in the total ROS rate, lipoperoxidation, protein carbonyl and DNA damage, when compared with senescent ASCs untreated. The guaraná also was able to modulate the activity and expression of antioxidants enzymes, increasing the catalase, decreasing the total SOD and a light decrease in the glutathione peroxidase was observed. Therefore, it can be said that the acute exposition of no senescent ASCs to H2O2 was highly cytotoxic, demonstrating the high sensibility of these cells to this stressor agent. However, senescent ASCs treated with guaraná showed the potential effect of this extract in the reversion of the cellular senescence. |
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2014-09-232014-09-232014-01-22MACHADO, Alencar Kolinski. Cito-genomic effect of hydrogen peroxide and guaraná (Paullinia cupana) in mesenchymal stem cells. 2014. 97 f. Dissertação (Mestrado em Farmácia) - Universidade Federal de Santa Maria, Santa Maria, 2014.http://repositorio.ufsm.br/handle/1/8989With advances in techniques of cell culture, stem cells (SCs) have to be used as an alternative therapy, firming the called regenerative medicine. An type of SC that has been studied is the Adipose-derived stem cell (ASCs) having identical characteristics of Mesenchymal Stem Cells (MSCs). To realize a transplanting of SCs, is necessary a large amount of cells, which makes such cultures are maintained in vitro for long periods of time, consequently these cells, with the repeated passages, enter cellular senescence, which prevents their use. Thus, some studies showed that the hydrogen peroxide (H2O2) can act to potentiate the proliferation of these cells. However, it is know that the oxidative metabolism can be associated with the senescence process. From this, the aim of this study was available the cito-genomic effect of the exposition of ASCs to H2O2 and to guaraná extract. Was realized isolation and cultivation of ASCs, that were, in forth passage, exposed to concentrations of 1-1000 μM of H2O2 and realized assays to available the cell viability, oxidative stress parameters, DNA damage and the apoptotic way of caspases. Moreover, senecents ASCs were treated with concentrations of 1-20 mg/mL of guaraná extract, being tested to the senescence reversion, cell viability, oxidative parameters, DNA damage and activity and expression of antioxidants enzymes. The results showed that the H2O2 cause cito-genomic damages to ASCs, with decrease in the cell viability mainly at the concentration of 200 μM, as well as increase the total reactive oxygen species (ROS) rate and lipidic peroxidation effect. The H2O2 also decreased the catalase activity in dose-dependent form e acted in the SOD activity. The treatments also conducted to a progressive increase of DNA damage. Moreover, in the eighth passage was observed the senescence of ASCs and the treatment with guaraná showed capacity to revert this phenotype in the concentration of 5 mg/mL, with reduction in the total ROS rate, lipoperoxidation, protein carbonyl and DNA damage, when compared with senescent ASCs untreated. The guaraná also was able to modulate the activity and expression of antioxidants enzymes, increasing the catalase, decreasing the total SOD and a light decrease in the glutathione peroxidase was observed. Therefore, it can be said that the acute exposition of no senescent ASCs to H2O2 was highly cytotoxic, demonstrating the high sensibility of these cells to this stressor agent. However, senescent ASCs treated with guaraná showed the potential effect of this extract in the reversion of the cellular senescence.Com os avanços das técnicas de cultivo celular, as células tronco (CTs) passaram a ser utilizadas como forma de terapia alternativa, firmando a chamada medicina regenerativa. Um tipo de CT que vem sendo estudada são as Células Tronco derivada do tecido adiposo (ASCs) que possuem características idênticas as Células Tronco Mesenquimais (CTMs). Para realizar um transplante de CTs, se faz necessária uma grande quantidade de células, o que faz com que tais cultivos sejam mantidos in vitro por longos períodos de tempo, consequentemente tais células, com as repetidas passagens, entram em senescência celular, o que inviabiliza a sua utilização. Sendo assim, algumas pesquisas demonstraram que o peróxido de hidrogênio (H2O2) pode atuar potencializando a proliferação destas células. Contudo, sabe-se que o metabolismo oxidativo está intimamente associado ao processo de senescência celular. A partir disto, o objetivo deste trabalho foi avaliar o efeito cito-genômico da exposição de ASCs ao H2O2 e ao extrato de guaraná. Foi realizado isolamento e cultivo de ASCs, as quais em quarta passagem foram expostas as concentrações de 1-1000 μM de H2O2 e realizados ensaios para a avaliação da viabilidade celular, parâmetros de estresse oxidativo, danos ao DNA e rota apoptótica das caspases. Além disso, ASCs senescentes foram tratadas com concentrações de 1-20mg/mL de extrato de guaraná, sendo testadas para a reversão da senescência, viabilidade celular, parâmetros oxidativos, de danos ao DNA e atividade e expressão de enzimas antioxidantes. Os resultados mostraram que o H2O2 causa danos cito-genômicos as ASCs, havendo redução da viabilidade celular principalmente a partir da concentração de 200 μM, assim como aumento da taxa total de espécies reativas de oxigênio (EROs) e peroxidação lipídica. O H2O2 também diminuiu a atividade da catalase de forma dose-dependente e atuou sobre a atividade da SOD. Os tratamentos também conduziram a um aumento progressivo dos danos ao DNA. Além disso, na oitava passagem foi observada a senescência das ASCs e o tratamento com guaraná mostrou capacidade de reversão deste fenótipo na concentração de 5 mg/mL, de forma a reduzir a taxa total de EROS, a lipoperoxidação, a carbonilação de proteínas e os danos ao DNA, quando comparado às ASCs não tratadas. O guaraná também foi capaz de modular a atividade e expressão de enzimas antioxidantes, aumentando a catalase, reduzindo a SOD total e uma leve redução na expressão da glutationa peroxidase. Logo, pode-se dizer que a exposição aguda de ASCs não senescentes ao H2O2 foi altamente citotóxica, demonstrando a alta sensibilidade destas células a este agente estressor. Todavia, ASCs senescentes tratadas com guaraná demonstraram o potencial efeito deste extrato na reversão da senescência celular.Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal de Santa MariaPrograma de Pós-Graduação em FarmacologiaUFSMBRFarmacologiaCélulas tronco derivadas do tecido adiposoMetabolismo oxidativoDanos celularesAdipose-derived stem cellsOxidative metabolismCellular damagesCNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIAEfeito cito-genômico do peróxido de hidrogênio e do guaraná (Paullinia cupana) em células tronco mesenquimaisCito-genomic effect of hydrogen peroxide and guaraná (Paullinia cupana) in mesenchymal stem cellsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisCruz, Ivana Beatrice Mânica dahttp://lattes.cnpq.br/3426369324110716Moresco, Rafael Noalhttp://lattes.cnpq.br/2269922709577261Unfer, Taís Cristinahttp://lattes.cnpq.br/6828277686285810http://lattes.cnpq.br/7339960872140748Machado, Alencar Kolinski2010000000004005003005003000b5fcfe0-b017-4163-80e2-17c110fbcf01ae6beeac-a53a-42d1-958d-a4151ee1378b05b536c5-19fb-4cae-9d69-46730a16ed098dca0fc1-540f-4d05-b047-8a344e8f08f0info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALMACHADO, ALENCAR KOLINSKI.pdfapplication/pdf1706014http://repositorio.ufsm.br/bitstream/1/8989/1/MACHADO%2c%20ALENCAR%20KOLINSKI.pdf407e365c94059e3f73e319b46e3b0f03MD51TEXTMACHADO, ALENCAR KOLINSKI.pdf.txtMACHADO, ALENCAR KOLINSKI.pdf.txtExtracted texttext/plain157362http://repositorio.ufsm.br/bitstream/1/8989/2/MACHADO%2c%20ALENCAR%20KOLINSKI.pdf.txtbf6baebfbfe3aacc24d52965927df9d7MD52THUMBNAILMACHADO, ALENCAR KOLINSKI.pdf.jpgMACHADO, ALENCAR KOLINSKI.pdf.jpgIM Thumbnailimage/jpeg4932http://repositorio.ufsm.br/bitstream/1/8989/3/MACHADO%2c%20ALENCAR%20KOLINSKI.pdf.jpg6d9f182c64e917bbfc654464bb549db9MD531/89892022-01-07 16:11:28.381oai:repositorio.ufsm.br:1/8989Repositório Institucionalhttp://repositorio.ufsm.br/PUBhttp://repositorio.ufsm.br/oai/requestopendoar:39132022-01-07T19:11:28Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.por.fl_str_mv |
Efeito cito-genômico do peróxido de hidrogênio e do guaraná (Paullinia cupana) em células tronco mesenquimais |
dc.title.alternative.eng.fl_str_mv |
Cito-genomic effect of hydrogen peroxide and guaraná (Paullinia cupana) in mesenchymal stem cells |
title |
Efeito cito-genômico do peróxido de hidrogênio e do guaraná (Paullinia cupana) em células tronco mesenquimais |
spellingShingle |
Efeito cito-genômico do peróxido de hidrogênio e do guaraná (Paullinia cupana) em células tronco mesenquimais Machado, Alencar Kolinski Células tronco derivadas do tecido adiposo Metabolismo oxidativo Danos celulares Adipose-derived stem cells Oxidative metabolism Cellular damages CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
title_short |
Efeito cito-genômico do peróxido de hidrogênio e do guaraná (Paullinia cupana) em células tronco mesenquimais |
title_full |
Efeito cito-genômico do peróxido de hidrogênio e do guaraná (Paullinia cupana) em células tronco mesenquimais |
title_fullStr |
Efeito cito-genômico do peróxido de hidrogênio e do guaraná (Paullinia cupana) em células tronco mesenquimais |
title_full_unstemmed |
Efeito cito-genômico do peróxido de hidrogênio e do guaraná (Paullinia cupana) em células tronco mesenquimais |
title_sort |
Efeito cito-genômico do peróxido de hidrogênio e do guaraná (Paullinia cupana) em células tronco mesenquimais |
author |
Machado, Alencar Kolinski |
author_facet |
Machado, Alencar Kolinski |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Cruz, Ivana Beatrice Mânica da |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/3426369324110716 |
dc.contributor.referee1.fl_str_mv |
Moresco, Rafael Noal |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/2269922709577261 |
dc.contributor.referee2.fl_str_mv |
Unfer, Taís Cristina |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/6828277686285810 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/7339960872140748 |
dc.contributor.author.fl_str_mv |
Machado, Alencar Kolinski |
contributor_str_mv |
Cruz, Ivana Beatrice Mânica da Moresco, Rafael Noal Unfer, Taís Cristina |
dc.subject.por.fl_str_mv |
Células tronco derivadas do tecido adiposo Metabolismo oxidativo Danos celulares |
topic |
Células tronco derivadas do tecido adiposo Metabolismo oxidativo Danos celulares Adipose-derived stem cells Oxidative metabolism Cellular damages CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
dc.subject.eng.fl_str_mv |
Adipose-derived stem cells Oxidative metabolism Cellular damages |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
description |
With advances in techniques of cell culture, stem cells (SCs) have to be used as an alternative therapy, firming the called regenerative medicine. An type of SC that has been studied is the Adipose-derived stem cell (ASCs) having identical characteristics of Mesenchymal Stem Cells (MSCs). To realize a transplanting of SCs, is necessary a large amount of cells, which makes such cultures are maintained in vitro for long periods of time, consequently these cells, with the repeated passages, enter cellular senescence, which prevents their use. Thus, some studies showed that the hydrogen peroxide (H2O2) can act to potentiate the proliferation of these cells. However, it is know that the oxidative metabolism can be associated with the senescence process. From this, the aim of this study was available the cito-genomic effect of the exposition of ASCs to H2O2 and to guaraná extract. Was realized isolation and cultivation of ASCs, that were, in forth passage, exposed to concentrations of 1-1000 μM of H2O2 and realized assays to available the cell viability, oxidative stress parameters, DNA damage and the apoptotic way of caspases. Moreover, senecents ASCs were treated with concentrations of 1-20 mg/mL of guaraná extract, being tested to the senescence reversion, cell viability, oxidative parameters, DNA damage and activity and expression of antioxidants enzymes. The results showed that the H2O2 cause cito-genomic damages to ASCs, with decrease in the cell viability mainly at the concentration of 200 μM, as well as increase the total reactive oxygen species (ROS) rate and lipidic peroxidation effect. The H2O2 also decreased the catalase activity in dose-dependent form e acted in the SOD activity. The treatments also conducted to a progressive increase of DNA damage. Moreover, in the eighth passage was observed the senescence of ASCs and the treatment with guaraná showed capacity to revert this phenotype in the concentration of 5 mg/mL, with reduction in the total ROS rate, lipoperoxidation, protein carbonyl and DNA damage, when compared with senescent ASCs untreated. The guaraná also was able to modulate the activity and expression of antioxidants enzymes, increasing the catalase, decreasing the total SOD and a light decrease in the glutathione peroxidase was observed. Therefore, it can be said that the acute exposition of no senescent ASCs to H2O2 was highly cytotoxic, demonstrating the high sensibility of these cells to this stressor agent. However, senescent ASCs treated with guaraná showed the potential effect of this extract in the reversion of the cellular senescence. |
publishDate |
2014 |
dc.date.accessioned.fl_str_mv |
2014-09-23 |
dc.date.available.fl_str_mv |
2014-09-23 |
dc.date.issued.fl_str_mv |
2014-01-22 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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masterThesis |
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publishedVersion |
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MACHADO, Alencar Kolinski. Cito-genomic effect of hydrogen peroxide and guaraná (Paullinia cupana) in mesenchymal stem cells. 2014. 97 f. Dissertação (Mestrado em Farmácia) - Universidade Federal de Santa Maria, Santa Maria, 2014. |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/8989 |
identifier_str_mv |
MACHADO, Alencar Kolinski. Cito-genomic effect of hydrogen peroxide and guaraná (Paullinia cupana) in mesenchymal stem cells. 2014. 97 f. Dissertação (Mestrado em Farmácia) - Universidade Federal de Santa Maria, Santa Maria, 2014. |
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http://repositorio.ufsm.br/handle/1/8989 |
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Universidade Federal de Santa Maria |
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UFSM |
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Farmacologia |
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