Envolvimento da enzima naphoxidase no efeito nefroprotetor do (-)-α-Bisabolol em um modelo de lesão renal aguda por isquemia-reperfusão In vitro
Ano de defesa: | 2018 |
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Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | |
Tipo de documento: | Tese |
Tipo de acesso: | Acesso aberto |
Idioma: | por |
Instituição de defesa: |
Não Informado pela instituição
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Programa de Pós-Graduação: |
Não Informado pela instituição
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Departamento: |
Não Informado pela instituição
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País: |
Não Informado pela instituição
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Palavras-chave em Português: | |
Link de acesso: | http://www.repositorio.ufc.br/handle/riufc/35867 |
Resumo: | Acute Renal Injury (AKI) is associated with high morbidity and mortality, generating public health costs. Ischemia and reperfusion (I/R) are events that frequently lead to AKI due to tissue hypoxia, reactive oxygen species (ROS) production, oxidative stress and programmed cell death by apoptosis. Previously, we have demonstrated the nephroprotective potential of (-)-α-Bisabolol, an unsaturated monocyclic sesquiterpene of chamomile flowers. The objective of the present study was to evaluate the mechanism of nephroprotection mediated by Bisabolol in human tubular renal cells after induction of I/R in an in vitro model. Additionally, this work intends to analyze the involvement of ROS generated by NOX4 in the lesion and cellular recovery. HK2 cells were exposed to I/R conditions and treated with Bisabolol at various concentrations. Cell viability was performed by the MTT assay. Treated and untreated cells were submitted to flow cytometry to evaluate necrotic/apoptotic cells, ROS production and mitochondrial transmembrane potential. TBARS and GSH were used as redox balance parameters. In addition, the levels of KIM-1 released in the supernatant were measured by ELISA. In order to identify an interaction between Bisabolol and NOX4, molecular docking and enzymatic assays in vitro were performed. Expression of the NOX4 isoform in treated cells was assessed by Western blotting. Finally, the cells were visualized by scanning electron microscopy. Bisabolol improved cell viability and prevented cell death by apoptosis, also indicated by decreased levels of KIM-1 in the cell supernatant. A decrease in ROS production and mitochondrial depolarization was observed, with antioxidant regulation by the increase of GSH and decrease of lipid peroxidation. It has also been shown that treatment with Bisabolol can inhibit NOX4. Scanning electron microscopy images showed that Bisabolol reduced cell injury induced by I/R. Therefore, treatment with Bisabolol protects renal tubular epithelial HK2 cells against the oxidative damage caused by I/R. This effect is related to inhibition of apoptosis, decreased release of KIM-1, and establishment of REDOX balance through inhibition of NOX4 activity in tubular cells. |
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Envolvimento da enzima naphoxidase no efeito nefroprotetor do (-)-α-Bisabolol em um modelo de lesão renal aguda por isquemia-reperfusão In vitroInvolvement of the enzyme NADPH oxidase in the nephroprotective effect of (-) - alpha-Bisabolol in a model of acute renal injury by ischemia-reperfusion in vitroIsquemiaReperfusãoNADPH OxidasesNADPH Oxidase 4Acute Renal Injury (AKI) is associated with high morbidity and mortality, generating public health costs. Ischemia and reperfusion (I/R) are events that frequently lead to AKI due to tissue hypoxia, reactive oxygen species (ROS) production, oxidative stress and programmed cell death by apoptosis. Previously, we have demonstrated the nephroprotective potential of (-)-α-Bisabolol, an unsaturated monocyclic sesquiterpene of chamomile flowers. The objective of the present study was to evaluate the mechanism of nephroprotection mediated by Bisabolol in human tubular renal cells after induction of I/R in an in vitro model. Additionally, this work intends to analyze the involvement of ROS generated by NOX4 in the lesion and cellular recovery. HK2 cells were exposed to I/R conditions and treated with Bisabolol at various concentrations. Cell viability was performed by the MTT assay. Treated and untreated cells were submitted to flow cytometry to evaluate necrotic/apoptotic cells, ROS production and mitochondrial transmembrane potential. TBARS and GSH were used as redox balance parameters. In addition, the levels of KIM-1 released in the supernatant were measured by ELISA. In order to identify an interaction between Bisabolol and NOX4, molecular docking and enzymatic assays in vitro were performed. Expression of the NOX4 isoform in treated cells was assessed by Western blotting. Finally, the cells were visualized by scanning electron microscopy. Bisabolol improved cell viability and prevented cell death by apoptosis, also indicated by decreased levels of KIM-1 in the cell supernatant. A decrease in ROS production and mitochondrial depolarization was observed, with antioxidant regulation by the increase of GSH and decrease of lipid peroxidation. It has also been shown that treatment with Bisabolol can inhibit NOX4. Scanning electron microscopy images showed that Bisabolol reduced cell injury induced by I/R. Therefore, treatment with Bisabolol protects renal tubular epithelial HK2 cells against the oxidative damage caused by I/R. This effect is related to inhibition of apoptosis, decreased release of KIM-1, and establishment of REDOX balance through inhibition of NOX4 activity in tubular cells.A Lesão Renal Aguda (LRA) está associada a alta morbidade e mortalidade, gerando custos de saúde pública. Isquemia e reperfusão (I/R) são eventos que frequentemente levam à LRA por hipóxia tecidual, produção de espécies reativas de oxigênio (ERO), estresse oxidativo e morte celular programada por apoptose. Anteriormente, demonstramos o potencial nefroprotetor do (-)-α-Bisabolol, um sesquiterpeno monocíclico insaturado de flores de camomila. O objetivo do presente trabalho foi avaliar o mecanismo de nefroproteção mediada pelo Bisabolol em células renais tubulares humanas após indução de I/R em um modelo in vitro. Adicionalmente, este trabalho pretende analisar o envolvimento de ERO gerados por NOX4 na lesão e recuperação celular. Células HK2 foram expostas a condições de I/R e tratadas com Bisabolol em várias concentrações. A viabilidade celular foi realizada pelo ensaio MTT. Células tratadas e não tratadas foram submetidas à citometria de fluxo para avaliar células necróticas/apoptóticas, produção de ERO e o potencial transmembrânico mitocondrial. TBARS e GSH foram utilizados como parâmetros do balanço redox. Além disso, os níveis de KIM-1 liberados no sobrenadante foram medidos por ELISA. A fim de identificar uma interação entre Bisabolol e NOX4, foram realizados ensaios de docking molecular e enzimáticos in vitro. A expressão da isoforma NOX4 em células tratadas foi avaliada por "Western-blot". Finalmente, as células foram visualizadas por microscopia eletrônica de varredura. O Bisabolol melhorou a viabilidade celular e preveniu a morte celular por apoptose, indicada também pela diminuição dos níveis de KIM-1 no sobrenadante celular. Foi observada uma diminuição na produção de ERO e despolarização mitocondrial, com regulação antioxidante pelo aumento da GSH e diminuição da peroxidação lipídica. Também foi demonstrado que o tratamento com Bisabolol pode inibir o NOX4. Imagens de microscopia eletrônica de varredura mostraram que o Bisabolol reduziu o dano celular induzido por I/R. Portanto, o tratamento com Bisabolol protege as células HK2 epiteliais tubulares renais contra o dano oxidativo ocasionado pela I/R. Este efeito está relacionado à inibição da apoptose, diminuição da liberação de KIM-1, e estabelecimento do equilíbrio REDOX através da inibição da atividade da NOX4 nas células tubulares.Martins, Alice Maria CostaSampaio, Tiago Lima2018-09-24T16:24:19Z2018-09-24T16:24:19Z2018-08-14info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfSAMPAIO, T. L. Envolvimento da enzima naphoxidase no efeito nefroprotetor do (-)-α-Bisabolol em um modelo de lesão renal aguda por isquemia-reperfusão In vitro. 2018. 102 f. Tese (Doutorado em Farmacologia) - Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2018.http://www.repositorio.ufc.br/handle/riufc/35867porreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccess2019-10-23T14:53:31Zoai:repositorio.ufc.br:riufc/35867Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-09-11T18:52:53.466416Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false |
dc.title.none.fl_str_mv |
Envolvimento da enzima naphoxidase no efeito nefroprotetor do (-)-α-Bisabolol em um modelo de lesão renal aguda por isquemia-reperfusão In vitro Involvement of the enzyme NADPH oxidase in the nephroprotective effect of (-) - alpha-Bisabolol in a model of acute renal injury by ischemia-reperfusion in vitro |
title |
Envolvimento da enzima naphoxidase no efeito nefroprotetor do (-)-α-Bisabolol em um modelo de lesão renal aguda por isquemia-reperfusão In vitro |
spellingShingle |
Envolvimento da enzima naphoxidase no efeito nefroprotetor do (-)-α-Bisabolol em um modelo de lesão renal aguda por isquemia-reperfusão In vitro Sampaio, Tiago Lima Isquemia Reperfusão NADPH Oxidases NADPH Oxidase 4 |
title_short |
Envolvimento da enzima naphoxidase no efeito nefroprotetor do (-)-α-Bisabolol em um modelo de lesão renal aguda por isquemia-reperfusão In vitro |
title_full |
Envolvimento da enzima naphoxidase no efeito nefroprotetor do (-)-α-Bisabolol em um modelo de lesão renal aguda por isquemia-reperfusão In vitro |
title_fullStr |
Envolvimento da enzima naphoxidase no efeito nefroprotetor do (-)-α-Bisabolol em um modelo de lesão renal aguda por isquemia-reperfusão In vitro |
title_full_unstemmed |
Envolvimento da enzima naphoxidase no efeito nefroprotetor do (-)-α-Bisabolol em um modelo de lesão renal aguda por isquemia-reperfusão In vitro |
title_sort |
Envolvimento da enzima naphoxidase no efeito nefroprotetor do (-)-α-Bisabolol em um modelo de lesão renal aguda por isquemia-reperfusão In vitro |
author |
Sampaio, Tiago Lima |
author_facet |
Sampaio, Tiago Lima |
author_role |
author |
dc.contributor.none.fl_str_mv |
Martins, Alice Maria Costa |
dc.contributor.author.fl_str_mv |
Sampaio, Tiago Lima |
dc.subject.por.fl_str_mv |
Isquemia Reperfusão NADPH Oxidases NADPH Oxidase 4 |
topic |
Isquemia Reperfusão NADPH Oxidases NADPH Oxidase 4 |
description |
Acute Renal Injury (AKI) is associated with high morbidity and mortality, generating public health costs. Ischemia and reperfusion (I/R) are events that frequently lead to AKI due to tissue hypoxia, reactive oxygen species (ROS) production, oxidative stress and programmed cell death by apoptosis. Previously, we have demonstrated the nephroprotective potential of (-)-α-Bisabolol, an unsaturated monocyclic sesquiterpene of chamomile flowers. The objective of the present study was to evaluate the mechanism of nephroprotection mediated by Bisabolol in human tubular renal cells after induction of I/R in an in vitro model. Additionally, this work intends to analyze the involvement of ROS generated by NOX4 in the lesion and cellular recovery. HK2 cells were exposed to I/R conditions and treated with Bisabolol at various concentrations. Cell viability was performed by the MTT assay. Treated and untreated cells were submitted to flow cytometry to evaluate necrotic/apoptotic cells, ROS production and mitochondrial transmembrane potential. TBARS and GSH were used as redox balance parameters. In addition, the levels of KIM-1 released in the supernatant were measured by ELISA. In order to identify an interaction between Bisabolol and NOX4, molecular docking and enzymatic assays in vitro were performed. Expression of the NOX4 isoform in treated cells was assessed by Western blotting. Finally, the cells were visualized by scanning electron microscopy. Bisabolol improved cell viability and prevented cell death by apoptosis, also indicated by decreased levels of KIM-1 in the cell supernatant. A decrease in ROS production and mitochondrial depolarization was observed, with antioxidant regulation by the increase of GSH and decrease of lipid peroxidation. It has also been shown that treatment with Bisabolol can inhibit NOX4. Scanning electron microscopy images showed that Bisabolol reduced cell injury induced by I/R. Therefore, treatment with Bisabolol protects renal tubular epithelial HK2 cells against the oxidative damage caused by I/R. This effect is related to inhibition of apoptosis, decreased release of KIM-1, and establishment of REDOX balance through inhibition of NOX4 activity in tubular cells. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-09-24T16:24:19Z 2018-09-24T16:24:19Z 2018-08-14 |
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 |
SAMPAIO, T. L. Envolvimento da enzima naphoxidase no efeito nefroprotetor do (-)-α-Bisabolol em um modelo de lesão renal aguda por isquemia-reperfusão In vitro. 2018. 102 f. Tese (Doutorado em Farmacologia) - Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2018. http://www.repositorio.ufc.br/handle/riufc/35867 |
identifier_str_mv |
SAMPAIO, T. L. Envolvimento da enzima naphoxidase no efeito nefroprotetor do (-)-α-Bisabolol em um modelo de lesão renal aguda por isquemia-reperfusão In vitro. 2018. 102 f. Tese (Doutorado em Farmacologia) - Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2018. |
url |
http://www.repositorio.ufc.br/handle/riufc/35867 |
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 |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da Universidade Federal do Ceará (UFC) instname:Universidade Federal do Ceará (UFC) instacron:UFC |
instname_str |
Universidade Federal do Ceará (UFC) |
instacron_str |
UFC |
institution |
UFC |
reponame_str |
Repositório Institucional da Universidade Federal do Ceará (UFC) |
collection |
Repositório Institucional da Universidade Federal do Ceará (UFC) |
repository.name.fl_str_mv |
Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC) |
repository.mail.fl_str_mv |
bu@ufc.br || repositorio@ufc.br |
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1813027802655490048 |