Consequências da modulação farmacológica da Na+, K+ - ATPase na epilepsia induzida por pilocarpina
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/15141 |
Resumo: | Epilepsy is one of the most common chronic neurological diseases worldwide, and despite a variety of drugs introduced into clinical practice, one-third of patients remain unresponsive to therapies. In addition, when considering the adverse effects therapies, epilepsy comorbidities, socioeconomic costs for public health, and the need for understanding molecular mechanisms involved in the disease, we conclude that epilepsy research is of fundamental importance. Na+, K+ -ATPase (NKA) is an important enzyme in the regulation of neuronal excitability, and an interesting pharmacological target to be explored in epilepsy. Evidence indicates that impairment in NKA activity contributes to epileptic seizures in mice and in humans with epilepsy. In order to evaluate new pharmacological tools that modulate NKA activity, the present study sought to identify some effects of the DRSSAb antibody, capable of activating NKA activity, and agrin, a protein that appears to be associated with inhibitory effects on NKA, on a experimental model of epilepsy. To this purpose, male C57BL/6 mice were subjected to status epilepticus (SE) by administration of repeated low doses of pilocarpine, and after a period these animals had spontaneous and recurrent seizures. In the in vitro experiments the hippocampal slices of control and epileptic mice were treated with DRSSAb. Glucose uptake improved by approximately 30% in the slices of epileptic animals after treatment with DRSSAb. Glutamate release increased by 83% in slices from epileptic animals, and returned to levels of control animals after treatment with DRSSAb. These results were obtained without interfering with cell viability. In the in vivo experiments, the intrahippocampal administration of DRSSAb restored crossing activity in the open field test; the number of crossings decreased in epileptic animals, and returned to the levels of the control group after DRSSAb treatment. Regarding agrin, the levels of protein expression after SE in hippocampus of mice were not altered 24 hours after SE. However, 14 days after SE there was a decrease in agrin expression, followed by an 71 % increase 60 days after SE. Moreover, the intrahippocampal administration of agrin increased the susceptibility to epileptic seizures in SE animals, but not in control mice. In summary, present data support the hypothesis that DRSSAb antibody and agrin are promising therapeutic strategies in the search for new treatments for epilepsy. |
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Consequências da modulação farmacológica da Na+, K+ - ATPase na epilepsia induzida por pilocarpinaConsequences of pharmacological modulation of Na+, K+ - ATPase in pilocarpine-induced epilepsyEpilepsiaNa+, K+-ATPaseDRSSAbAgrinaHiperexcitabilidadeExcitotoxicidadeEpilepsyHyperexcitabilityExcitotoxicityCNPQ::CIENCIAS DA SAUDE::FARMACIAEpilepsy is one of the most common chronic neurological diseases worldwide, and despite a variety of drugs introduced into clinical practice, one-third of patients remain unresponsive to therapies. In addition, when considering the adverse effects therapies, epilepsy comorbidities, socioeconomic costs for public health, and the need for understanding molecular mechanisms involved in the disease, we conclude that epilepsy research is of fundamental importance. Na+, K+ -ATPase (NKA) is an important enzyme in the regulation of neuronal excitability, and an interesting pharmacological target to be explored in epilepsy. Evidence indicates that impairment in NKA activity contributes to epileptic seizures in mice and in humans with epilepsy. In order to evaluate new pharmacological tools that modulate NKA activity, the present study sought to identify some effects of the DRSSAb antibody, capable of activating NKA activity, and agrin, a protein that appears to be associated with inhibitory effects on NKA, on a experimental model of epilepsy. To this purpose, male C57BL/6 mice were subjected to status epilepticus (SE) by administration of repeated low doses of pilocarpine, and after a period these animals had spontaneous and recurrent seizures. In the in vitro experiments the hippocampal slices of control and epileptic mice were treated with DRSSAb. Glucose uptake improved by approximately 30% in the slices of epileptic animals after treatment with DRSSAb. Glutamate release increased by 83% in slices from epileptic animals, and returned to levels of control animals after treatment with DRSSAb. These results were obtained without interfering with cell viability. In the in vivo experiments, the intrahippocampal administration of DRSSAb restored crossing activity in the open field test; the number of crossings decreased in epileptic animals, and returned to the levels of the control group after DRSSAb treatment. Regarding agrin, the levels of protein expression after SE in hippocampus of mice were not altered 24 hours after SE. However, 14 days after SE there was a decrease in agrin expression, followed by an 71 % increase 60 days after SE. Moreover, the intrahippocampal administration of agrin increased the susceptibility to epileptic seizures in SE animals, but not in control mice. In summary, present data support the hypothesis that DRSSAb antibody and agrin are promising therapeutic strategies in the search for new treatments for epilepsy.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESA epilepsia é uma das doenças neurológicas crônicas mais comum em todo o mundo, e apesar da diversidade de medicamentos disponíveis na prática clínica, um terço dos pacientes continuam não respondendo as terapias. Além disso, ao considerarmos os efeitos adversos das terapias disponíveis, as comorbidades da epilepsia, os custos socioeconômicos para saúde pública e a necessidade de elucidação dos mecanismos moleculares que envolvem a doença, concluímos que há grande necessidade do contínuo desenvolvimento das pesquisas em epilepsia. A Na+, K+-ATPase (NKA) é uma enzima importante na regulação da excitabilidade neuronal, e um interessante alvo farmacológico a ser explorado na epilepsia. Evidências indicam que o prejuízo na atividade da NKA contribui para crises epiléptica em camundongos e em humanos com epilepsia. A fim de avaliar novas ferramentas farmacológicas que modulem a atividade da NKA, o presente estudo procurou identificar alguns efeitos do anticorpo DRSSAb, capaz de ativar a atividade da NKA, e da agrina, proteína que parece estar associada com efeitos inibitórios sobre a NKA, em um modelo experimental de epilepsia. Para isso, camundongos adultos machos C57BL/6 foram submetidos ao estado de mal epiléptico (SE) através da administração de repetidas doses de pilocarpina, e após um período esses animais passam a apresentar crises convulsivas espontâneas e recorrentes. Nos experimentos in vitro as fatias hipocampais de camundongos epilépticos e controles foram tratadas com DRSSAb. Obteve-se uma melhora na captação de glicose de aproximadamente 30% nas fatias dos animais epilépticos em contato com o DRSSAb. A liberação de glutamato aumentou em 83% nas fatias dos animais epilépticos, e diminuiu aos níveis dos animais controles com o tratamento do DRSSAb. Os resultados foram obtidos sem interferir na viabilidade celular. Nos experimentos in vivo, a administração intrahipocampal do DRSSAb reestabeleceu a atividade de cruzamentos no teste de campo aberto; o número de cruzamentos diminuiu a metade nos animais tratados com pilocarpina, e foram reestabelecidos aos níveis do grupo controle após tratamento com DRSSAb. Quanto a agrina, os níveis de expressão proteica após o SE em hipocampo de camundongos não foram alterados em 24 horas. Porém, 14 dias após o SE houve uma diminuição na expressão da agrina, seguida de um aumento em 71% 60 dias após o SE. A administração intrahipocampal de agrina aumentou a susceptibilidade a crises epilépticas em animais SE, mas não em camundongos controles. Em resumo, os dados desta tese dão suporte para a hipótese que o anticorpo DRSSAb e a agrinasão promissoras estratégias terapêuticas na busca de novos tratamentos para epilepsia.Universidade Federal de Santa MariaBrasilFarmacologiaUFSMPrograma de Pós-Graduação em FarmacologiaCentro de Ciências da SaúdeOliveira, Mauro Schneiderhttp://lattes.cnpq.br/7132934163734175Guerra, Gustavo Petrihttp://lattes.cnpq.br/8053094752538070Sari, Marcel Henrique Marcondeshttp://lattes.cnpq.br/2698465900773455Oliveira, Sara Marchesan dehttp://lattes.cnpq.br/6574555059806902Funck, Vinícius Rafaelhttp://lattes.cnpq.br/6401593292343704Freitas, Mayara Lutchemeyer de2018-12-19T18:22:57Z2018-12-19T18:22:57Z2018-08-24info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/15141porAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2018-12-20T05:02:12Zoai:repositorio.ufsm.br:1/15141Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2018-12-20T05:02:12Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.none.fl_str_mv |
Consequências da modulação farmacológica da Na+, K+ - ATPase na epilepsia induzida por pilocarpina Consequences of pharmacological modulation of Na+, K+ - ATPase in pilocarpine-induced epilepsy |
title |
Consequências da modulação farmacológica da Na+, K+ - ATPase na epilepsia induzida por pilocarpina |
spellingShingle |
Consequências da modulação farmacológica da Na+, K+ - ATPase na epilepsia induzida por pilocarpina Freitas, Mayara Lutchemeyer de Epilepsia Na+, K+-ATPase DRSSAb Agrina Hiperexcitabilidade Excitotoxicidade Epilepsy Hyperexcitability Excitotoxicity CNPQ::CIENCIAS DA SAUDE::FARMACIA |
title_short |
Consequências da modulação farmacológica da Na+, K+ - ATPase na epilepsia induzida por pilocarpina |
title_full |
Consequências da modulação farmacológica da Na+, K+ - ATPase na epilepsia induzida por pilocarpina |
title_fullStr |
Consequências da modulação farmacológica da Na+, K+ - ATPase na epilepsia induzida por pilocarpina |
title_full_unstemmed |
Consequências da modulação farmacológica da Na+, K+ - ATPase na epilepsia induzida por pilocarpina |
title_sort |
Consequências da modulação farmacológica da Na+, K+ - ATPase na epilepsia induzida por pilocarpina |
author |
Freitas, Mayara Lutchemeyer de |
author_facet |
Freitas, Mayara Lutchemeyer de |
author_role |
author |
dc.contributor.none.fl_str_mv |
Oliveira, Mauro Schneider http://lattes.cnpq.br/7132934163734175 Guerra, Gustavo Petri http://lattes.cnpq.br/8053094752538070 Sari, Marcel Henrique Marcondes http://lattes.cnpq.br/2698465900773455 Oliveira, Sara Marchesan de http://lattes.cnpq.br/6574555059806902 Funck, Vinícius Rafael http://lattes.cnpq.br/6401593292343704 |
dc.contributor.author.fl_str_mv |
Freitas, Mayara Lutchemeyer de |
dc.subject.por.fl_str_mv |
Epilepsia Na+, K+-ATPase DRSSAb Agrina Hiperexcitabilidade Excitotoxicidade Epilepsy Hyperexcitability Excitotoxicity CNPQ::CIENCIAS DA SAUDE::FARMACIA |
topic |
Epilepsia Na+, K+-ATPase DRSSAb Agrina Hiperexcitabilidade Excitotoxicidade Epilepsy Hyperexcitability Excitotoxicity CNPQ::CIENCIAS DA SAUDE::FARMACIA |
description |
Epilepsy is one of the most common chronic neurological diseases worldwide, and despite a variety of drugs introduced into clinical practice, one-third of patients remain unresponsive to therapies. In addition, when considering the adverse effects therapies, epilepsy comorbidities, socioeconomic costs for public health, and the need for understanding molecular mechanisms involved in the disease, we conclude that epilepsy research is of fundamental importance. Na+, K+ -ATPase (NKA) is an important enzyme in the regulation of neuronal excitability, and an interesting pharmacological target to be explored in epilepsy. Evidence indicates that impairment in NKA activity contributes to epileptic seizures in mice and in humans with epilepsy. In order to evaluate new pharmacological tools that modulate NKA activity, the present study sought to identify some effects of the DRSSAb antibody, capable of activating NKA activity, and agrin, a protein that appears to be associated with inhibitory effects on NKA, on a experimental model of epilepsy. To this purpose, male C57BL/6 mice were subjected to status epilepticus (SE) by administration of repeated low doses of pilocarpine, and after a period these animals had spontaneous and recurrent seizures. In the in vitro experiments the hippocampal slices of control and epileptic mice were treated with DRSSAb. Glucose uptake improved by approximately 30% in the slices of epileptic animals after treatment with DRSSAb. Glutamate release increased by 83% in slices from epileptic animals, and returned to levels of control animals after treatment with DRSSAb. These results were obtained without interfering with cell viability. In the in vivo experiments, the intrahippocampal administration of DRSSAb restored crossing activity in the open field test; the number of crossings decreased in epileptic animals, and returned to the levels of the control group after DRSSAb treatment. Regarding agrin, the levels of protein expression after SE in hippocampus of mice were not altered 24 hours after SE. However, 14 days after SE there was a decrease in agrin expression, followed by an 71 % increase 60 days after SE. Moreover, the intrahippocampal administration of agrin increased the susceptibility to epileptic seizures in SE animals, but not in control mice. In summary, present data support the hypothesis that DRSSAb antibody and agrin are promising therapeutic strategies in the search for new treatments for epilepsy. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-12-19T18:22:57Z 2018-12-19T18:22:57Z 2018-08-24 |
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 |
http://repositorio.ufsm.br/handle/1/15141 |
url |
http://repositorio.ufsm.br/handle/1/15141 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Farmacologia UFSM Programa de Pós-Graduação em Farmacologia Centro de Ciências da Saúde |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Farmacologia UFSM Programa de Pós-Graduação em Farmacologia Centro de Ciências da Saúde |
dc.source.none.fl_str_mv |
reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
institution |
UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
collection |
Manancial - Repositório Digital da UFSM |
repository.name.fl_str_mv |
Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
atendimento.sib@ufsm.br||tedebc@gmail.com |
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1805922061520994304 |