Efeito diferencial do diazepam sobre a atividade da enzima Na+,K+-ATPase no hipocampo e córtex entorrinal
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações do UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/11848 |
Resumo: | Na+,K+-ATPase is ubiquitously expressed in the plasma membrane of all animal cells where serves as the principal regulator of intracellular ion homeostasis. Na+,K+-ATPase activity is activated by Na+ and K+ and current evidence indicates that total Na+,K+-ATPase activity is, in general, inhibited by anions. However, the effect of pharmacologically-induced Cl- flux on α1- and α2/3-subunit containing Na+,K+-ATPase activity is not established. In this study we investigated the effect of diazepam, a GABAA receptor positive allosteric modulator, on α1- and α2/3-subunit containing Na+,K+-ATPase activity. Hippocampal and cortical slices were incubated with diazepam (0, 0.05, 0.15 or 0.5 μM) and/or flumazenil (0, 0.005, 0.015, 0.05, 0.15, 0.5 or 1.5 μM) for 10 minutes. After incubation the slices were homogenized and α1 and α2/3 Na+,K+-ATPase activity were assayed using ouabain 3 μM (that inhibits α2/3-subunit containing Na+,K+-ATPase) and 4 mM (that inhibits both isoforms). Diazepam caused a 50% decrease of α2/3-subunit containing Na+,K+-ATPase activity in the hippocampus, but did not alter enzyme activity in the entorhinal cortex. The effect of diazepam was prevented by flumazenil, indicating that the decrease of Na+,K+-ATPase was involved GABAA receptors. Furthermore, a low chloride medium abolished the diazepam-induced decrease of Na+,K+-ATPase activity. Our data suggests that Na+,K+-ATPase in the hippocampus is sensitive to the pharmacological effects of a benzodiazepine by GABAA receptor-mediated mechanisms. Keywords: sodium pump. GABAA receptor. diazepam. flumazenil. chloride ion. hippocampus. entorhinal córtex. |
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2017-10-16T11:52:47Z2017-10-16T11:52:47Z2016-11-29http://repositorio.ufsm.br/handle/1/11848Na+,K+-ATPase is ubiquitously expressed in the plasma membrane of all animal cells where serves as the principal regulator of intracellular ion homeostasis. Na+,K+-ATPase activity is activated by Na+ and K+ and current evidence indicates that total Na+,K+-ATPase activity is, in general, inhibited by anions. However, the effect of pharmacologically-induced Cl- flux on α1- and α2/3-subunit containing Na+,K+-ATPase activity is not established. In this study we investigated the effect of diazepam, a GABAA receptor positive allosteric modulator, on α1- and α2/3-subunit containing Na+,K+-ATPase activity. Hippocampal and cortical slices were incubated with diazepam (0, 0.05, 0.15 or 0.5 μM) and/or flumazenil (0, 0.005, 0.015, 0.05, 0.15, 0.5 or 1.5 μM) for 10 minutes. After incubation the slices were homogenized and α1 and α2/3 Na+,K+-ATPase activity were assayed using ouabain 3 μM (that inhibits α2/3-subunit containing Na+,K+-ATPase) and 4 mM (that inhibits both isoforms). Diazepam caused a 50% decrease of α2/3-subunit containing Na+,K+-ATPase activity in the hippocampus, but did not alter enzyme activity in the entorhinal cortex. The effect of diazepam was prevented by flumazenil, indicating that the decrease of Na+,K+-ATPase was involved GABAA receptors. Furthermore, a low chloride medium abolished the diazepam-induced decrease of Na+,K+-ATPase activity. Our data suggests that Na+,K+-ATPase in the hippocampus is sensitive to the pharmacological effects of a benzodiazepine by GABAA receptor-mediated mechanisms. Keywords: sodium pump. GABAA receptor. diazepam. flumazenil. chloride ion. hippocampus. entorhinal córtex.A enzima Na+,K+-ATPase, ou bomba de sódio, é expressa na membrana plasmática de células eucarióticas, onde atua como principal regulador da homeostase iônica intracelular. A enzima Na+,K+-ATPase é ativada pelos íons Na+ and K+ e evidências indicam que a atividade total da enzima Na+,K+-ATPase é inibida por ânions. Entretanto, o efeito do fluxo de cloreto induzido farmacologicamente sobre a atividade das subunidades α1 e α2/3 da enzima Na+,K+-ATPase ainda não foi investigado. Neste estudo, nós investigamos o efeito do diazepam, um modulador alostérico positivo do receptor GABAA na atividade específica das subunidades α1 e α2/3 da Na+,K+-ATPase. Fatias de hipocampo e de córtex entorrinal foram incubadas com diazepam (0; 0,05; 0,15 ou 0,5 μM) e/ou flumazenil (0; 0,005, 0,015; 0,05; 0,15; 0,5 ou 1,5 μM) por 10 minutos. Após a incubação, as fatias foram homogeneizadas e a atividade das subunidades α1 e α2/3 da enzima Na+,K+-ATPase foi determinada. Diazepam diminuiu 50% a atividade da subunidade α2/3 da Na+,K+-ATPase no hipocampo, mas não alterou a atividade da enzima em córtex entorrinal. O efeito do diazepam foi prevenido por flumazenil, indicando que a diminuição da atividade da Na+,K+-ATPase envolveu a ativação dos receptores GABAA. Além disso, a baixa concentração de cloreto no meio de incubação aboliu a diminuição da atividade enzimática induzida por diazepam. Nossos dados sugerem que a enzima Na+,K+-ATPase no hipocampo é sensível a efeitos farmacológicos dos benzodiazepínicos por meio de mecanismos ativados por receptores GABAérgicos.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESporUniversidade Federal de Santa MariaCentro de Ciências da SaúdePrograma de Pós-Graduação em FarmacologiaUFSMBrasilFarmacologiaAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessBomba de sódioNa+,K+-ATPaseReceptor GABAADiazepamFlumazenilCloretoHipocampoCórtexSodium pumpGABAA receptorDiazepamFlumazenilChloride ionHippocampusEntorhinal córtexCNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIAEfeito diferencial do diazepam sobre a atividade da enzima Na+,K+-ATPase no hipocampo e córtex entorrinalDifferencial effect of diazepam on Na+,K+-ATPase activity in the hippocampus and entorhinal cortexinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisMello, Carlos Fernando dehttp://lattes.cnpq.br/3913887223894236Oliveira, Mauro Schneiderhttp://lattes.cnpq.br/7132934163734175Piato, Angelo Luis Stapassolihttp://lattes.cnpq.br/0837379287129794http://lattes.cnpq.br/9860511663248830Marafiga, Joseane Righes20100000000060060082e537b0-4a43-400e-9f82-0cd91952adeac29e7b1c-c886-45f8-8fec-22763c9a942e10d2124b-3909-4951-a54e-a167355aa75595833c5b-39f0-4813-8194-078183f725f2reponame:Biblioteca Digital de Teses e Dissertações do UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALMarafiga, Joseane Righes.pdfMarafiga, Joseane Righes.pdfDissertação de Mestradoapplication/pdf2070371http://repositorio.ufsm.br/bitstream/1/11848/1/Marafiga%2c%20Joseane%20Righes.pdf3226abdb2209e4e08110da587990a969MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; 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dc.title.por.fl_str_mv |
Efeito diferencial do diazepam sobre a atividade da enzima Na+,K+-ATPase no hipocampo e córtex entorrinal |
dc.title.alternative.eng.fl_str_mv |
Differencial effect of diazepam on Na+,K+-ATPase activity in the hippocampus and entorhinal cortex |
title |
Efeito diferencial do diazepam sobre a atividade da enzima Na+,K+-ATPase no hipocampo e córtex entorrinal |
spellingShingle |
Efeito diferencial do diazepam sobre a atividade da enzima Na+,K+-ATPase no hipocampo e córtex entorrinal Marafiga, Joseane Righes Bomba de sódio Na+,K+-ATPase Receptor GABAA Diazepam Flumazenil Cloreto Hipocampo Córtex Sodium pump GABAA receptor Diazepam Flumazenil Chloride ion Hippocampus Entorhinal córtex CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
title_short |
Efeito diferencial do diazepam sobre a atividade da enzima Na+,K+-ATPase no hipocampo e córtex entorrinal |
title_full |
Efeito diferencial do diazepam sobre a atividade da enzima Na+,K+-ATPase no hipocampo e córtex entorrinal |
title_fullStr |
Efeito diferencial do diazepam sobre a atividade da enzima Na+,K+-ATPase no hipocampo e córtex entorrinal |
title_full_unstemmed |
Efeito diferencial do diazepam sobre a atividade da enzima Na+,K+-ATPase no hipocampo e córtex entorrinal |
title_sort |
Efeito diferencial do diazepam sobre a atividade da enzima Na+,K+-ATPase no hipocampo e córtex entorrinal |
author |
Marafiga, Joseane Righes |
author_facet |
Marafiga, Joseane Righes |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Mello, Carlos Fernando de |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/3913887223894236 |
dc.contributor.referee1.fl_str_mv |
Oliveira, Mauro Schneider |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/7132934163734175 |
dc.contributor.referee2.fl_str_mv |
Piato, Angelo Luis Stapassoli |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/0837379287129794 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/9860511663248830 |
dc.contributor.author.fl_str_mv |
Marafiga, Joseane Righes |
contributor_str_mv |
Mello, Carlos Fernando de Oliveira, Mauro Schneider Piato, Angelo Luis Stapassoli |
dc.subject.por.fl_str_mv |
Bomba de sódio Na+,K+-ATPase Receptor GABAA Diazepam Flumazenil Cloreto Hipocampo Córtex |
topic |
Bomba de sódio Na+,K+-ATPase Receptor GABAA Diazepam Flumazenil Cloreto Hipocampo Córtex Sodium pump GABAA receptor Diazepam Flumazenil Chloride ion Hippocampus Entorhinal córtex CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
dc.subject.eng.fl_str_mv |
Sodium pump GABAA receptor Diazepam Flumazenil Chloride ion Hippocampus Entorhinal córtex |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
description |
Na+,K+-ATPase is ubiquitously expressed in the plasma membrane of all animal cells where serves as the principal regulator of intracellular ion homeostasis. Na+,K+-ATPase activity is activated by Na+ and K+ and current evidence indicates that total Na+,K+-ATPase activity is, in general, inhibited by anions. However, the effect of pharmacologically-induced Cl- flux on α1- and α2/3-subunit containing Na+,K+-ATPase activity is not established. In this study we investigated the effect of diazepam, a GABAA receptor positive allosteric modulator, on α1- and α2/3-subunit containing Na+,K+-ATPase activity. Hippocampal and cortical slices were incubated with diazepam (0, 0.05, 0.15 or 0.5 μM) and/or flumazenil (0, 0.005, 0.015, 0.05, 0.15, 0.5 or 1.5 μM) for 10 minutes. After incubation the slices were homogenized and α1 and α2/3 Na+,K+-ATPase activity were assayed using ouabain 3 μM (that inhibits α2/3-subunit containing Na+,K+-ATPase) and 4 mM (that inhibits both isoforms). Diazepam caused a 50% decrease of α2/3-subunit containing Na+,K+-ATPase activity in the hippocampus, but did not alter enzyme activity in the entorhinal cortex. The effect of diazepam was prevented by flumazenil, indicating that the decrease of Na+,K+-ATPase was involved GABAA receptors. Furthermore, a low chloride medium abolished the diazepam-induced decrease of Na+,K+-ATPase activity. Our data suggests that Na+,K+-ATPase in the hippocampus is sensitive to the pharmacological effects of a benzodiazepine by GABAA receptor-mediated mechanisms. Keywords: sodium pump. GABAA receptor. diazepam. flumazenil. chloride ion. hippocampus. entorhinal córtex. |
publishDate |
2016 |
dc.date.issued.fl_str_mv |
2016-11-29 |
dc.date.accessioned.fl_str_mv |
2017-10-16T11:52:47Z |
dc.date.available.fl_str_mv |
2017-10-16T11:52:47Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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masterThesis |
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http://repositorio.ufsm.br/handle/1/11848 |
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http://repositorio.ufsm.br/handle/1/11848 |
dc.language.iso.fl_str_mv |
por |
language |
por |
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201000000000 |
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600 600 |
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Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
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Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências da Saúde |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Farmacologia |
dc.publisher.initials.fl_str_mv |
UFSM |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
Farmacologia |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências da Saúde |
dc.source.none.fl_str_mv |
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