Hidrólise de atp e adp em líquor humano

Detalhes bibliográficos
Autor(a) principal: Sperotto, Rita Leal
Data de Publicação: 2008
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
dARK ID: ark:/26339/001300000w40s
Texto Completo: http://repositorio.ufsm.br/handle/1/11088
Resumo: Adenine nucleotides hydrolysis is an important step of the neuromodulation of CNS and some of its breakdown products are able to protect the nervous tissue against brain injuries. The activity of NTPDase (EC 3.6.1.5, apyrase, CD39) was verified in cerebrospinal fluid (CSF) from patients without neural inflammatory process under different conditions and in the presence of several inhibitors. The samples were chose considering the low protein levels, normal glucose levels, low leukocyte count and CSF differential count. We chose use the supernatant 1 (S1) for enzyme assay due to the best enzymatic activities in this fraction. The best hydrolyze temperature was 37°C to ATP and ADP. This enzyme was cation-dependent, with a maximal rate for ATP and ADP hydrolysis in pH 8.0 in the presence of 5mM Ca+2. Sodum azide inhibited both nucleotide hydrolysis at concentrations higher than 10 mM. Sodium fluoride inhibited the ATP and ADP hydrolysis at concentrations of 15 mM and 20 mM. The Na+ K+ ATPase inhibitor ouabain, did not affect ATP or ADP hydrolysis. The inhibitor P-type ATPase lanthanum 5mM was ineffective on ATPDase hydrolysis. Suramin (30-300 μM) inhibited ATP and ADP hydrolysis and presented a maximal inhibitory effect of 50% at 300 μM. The results of the present study demonstrated that ATP and ADP hydrolysis from human CSF presented a similar response those obtained from rat synaptosomes.
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spelling Hidrólise de atp e adp em líquor humanoAtp and adp hydrolysis in human cerebrospinal fluidNTPDaseCaracterizaçãoLíquido cefalorraquidianoNeuroproteçãoCSFNucleotidesNeuroprotectionCNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICAAdenine nucleotides hydrolysis is an important step of the neuromodulation of CNS and some of its breakdown products are able to protect the nervous tissue against brain injuries. The activity of NTPDase (EC 3.6.1.5, apyrase, CD39) was verified in cerebrospinal fluid (CSF) from patients without neural inflammatory process under different conditions and in the presence of several inhibitors. The samples were chose considering the low protein levels, normal glucose levels, low leukocyte count and CSF differential count. We chose use the supernatant 1 (S1) for enzyme assay due to the best enzymatic activities in this fraction. The best hydrolyze temperature was 37°C to ATP and ADP. This enzyme was cation-dependent, with a maximal rate for ATP and ADP hydrolysis in pH 8.0 in the presence of 5mM Ca+2. Sodum azide inhibited both nucleotide hydrolysis at concentrations higher than 10 mM. Sodium fluoride inhibited the ATP and ADP hydrolysis at concentrations of 15 mM and 20 mM. The Na+ K+ ATPase inhibitor ouabain, did not affect ATP or ADP hydrolysis. The inhibitor P-type ATPase lanthanum 5mM was ineffective on ATPDase hydrolysis. Suramin (30-300 μM) inhibited ATP and ADP hydrolysis and presented a maximal inhibitory effect of 50% at 300 μM. The results of the present study demonstrated that ATP and ADP hydrolysis from human CSF presented a similar response those obtained from rat synaptosomes.A análise do líquor é de grande importância para a detecção de desordens neurológicas de diversas etiologias. O ATP junto a seus produtos de hidrólise (ADP, AMP e adenosina) desempenha importantes funções junto ao SNC, as quais envolvem ações neuroprotetoras em doenças de etiologias variadas. O presente estudo tem como objetivos verificar a ocorrência de hidrólise de nucleotídeos da adenina em líquor de pacientes sem doença inflamatória do sistema nervoso. A determinação da atividade enzimática da NTPDase foi feita em líquor humano em diferentes condições experimentais e na presença de inibidores. As amostras foram escolhidas de acordo com os baixos níveis protéicos, valores normais de glicose e contagem celular diminuída. Foi escolhido o sobrenadante 1 (S1) por apresentar melhores atividades enzimáticas. A melhor temperatura de hidrólise do ATP e ADP foi 37°C. Esta enzima é cátion dependente, sendo a atividade de hidrólise ótima do ATP em presença de 5 mM Ca+2 e o ADP 5-7 mM de Ca+2, ambos em pH 8.0. A azida sódica alterou a atividade enzimática do ATP e do ADP somente nas concentrações mais altas deste inibidor da ATPase mitocondrial. A ouabaína, um inibidor da Na+/K+ ATPase não afetou a hidrólise do ATP/ADP. O inibidor de ATPase tipo-P lantânio (5 mM) foi ineficaz na hidrólise dos nucleotídeos. O suramin (30-300 XM), inibidor específico de NTPDase, inibiu a hidrólise do ATP/ADP e apresentou máximo efeito inibitório na concentração de 300 XM. Os resultados deste estudo mostraram que a hidrólise de ATP/ADP em líquor humano apresentou uma resposta semelhante àquelas obtidas em sinaptossomas de ratos.Universidade Federal de Santa MariaBRBioquímicaUFSMPrograma de Pós-Graduação em Ciências Biológicas: Bioquímica ToxicológicaLoro, Vania Luciahttp://lattes.cnpq.br/6392817606416780Schetinger, Maria Rosa ChitolinaPavanato, Maria Amáliahttp://lattes.cnpq.br/8701892865724171Sperotto, Rita Leal2017-04-262017-04-262008-03-27info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfapplication/pdfSPEROTTO, Rita Leal. Atp and adp hydrolysis in human cerebrospinal fluid. 2008. 61 f. Dissertação (Mestrado em Bioquímica) - Universidade Federal de Santa Maria, Santa Maria, 2008.http://repositorio.ufsm.br/handle/1/11088ark:/26339/001300000w40sporinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2022-08-25T19:35:28Zoai:repositorio.ufsm.br:1/11088Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2022-08-25T19:35:28Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv Hidrólise de atp e adp em líquor humano
Atp and adp hydrolysis in human cerebrospinal fluid
title Hidrólise de atp e adp em líquor humano
spellingShingle Hidrólise de atp e adp em líquor humano
Sperotto, Rita Leal
NTPDase
Caracterização
Líquido cefalorraquidiano
Neuroproteção
CSF
Nucleotides
Neuroprotection
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
title_short Hidrólise de atp e adp em líquor humano
title_full Hidrólise de atp e adp em líquor humano
title_fullStr Hidrólise de atp e adp em líquor humano
title_full_unstemmed Hidrólise de atp e adp em líquor humano
title_sort Hidrólise de atp e adp em líquor humano
author Sperotto, Rita Leal
author_facet Sperotto, Rita Leal
author_role author
dc.contributor.none.fl_str_mv Loro, Vania Lucia
http://lattes.cnpq.br/6392817606416780
Schetinger, Maria Rosa Chitolina
Pavanato, Maria Amália
http://lattes.cnpq.br/8701892865724171
dc.contributor.author.fl_str_mv Sperotto, Rita Leal
dc.subject.por.fl_str_mv NTPDase
Caracterização
Líquido cefalorraquidiano
Neuroproteção
CSF
Nucleotides
Neuroprotection
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
topic NTPDase
Caracterização
Líquido cefalorraquidiano
Neuroproteção
CSF
Nucleotides
Neuroprotection
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
description Adenine nucleotides hydrolysis is an important step of the neuromodulation of CNS and some of its breakdown products are able to protect the nervous tissue against brain injuries. The activity of NTPDase (EC 3.6.1.5, apyrase, CD39) was verified in cerebrospinal fluid (CSF) from patients without neural inflammatory process under different conditions and in the presence of several inhibitors. The samples were chose considering the low protein levels, normal glucose levels, low leukocyte count and CSF differential count. We chose use the supernatant 1 (S1) for enzyme assay due to the best enzymatic activities in this fraction. The best hydrolyze temperature was 37°C to ATP and ADP. This enzyme was cation-dependent, with a maximal rate for ATP and ADP hydrolysis in pH 8.0 in the presence of 5mM Ca+2. Sodum azide inhibited both nucleotide hydrolysis at concentrations higher than 10 mM. Sodium fluoride inhibited the ATP and ADP hydrolysis at concentrations of 15 mM and 20 mM. The Na+ K+ ATPase inhibitor ouabain, did not affect ATP or ADP hydrolysis. The inhibitor P-type ATPase lanthanum 5mM was ineffective on ATPDase hydrolysis. Suramin (30-300 μM) inhibited ATP and ADP hydrolysis and presented a maximal inhibitory effect of 50% at 300 μM. The results of the present study demonstrated that ATP and ADP hydrolysis from human CSF presented a similar response those obtained from rat synaptosomes.
publishDate 2008
dc.date.none.fl_str_mv 2008-03-27
2017-04-26
2017-04-26
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 SPEROTTO, Rita Leal. Atp and adp hydrolysis in human cerebrospinal fluid. 2008. 61 f. Dissertação (Mestrado em Bioquímica) - Universidade Federal de Santa Maria, Santa Maria, 2008.
http://repositorio.ufsm.br/handle/1/11088
dc.identifier.dark.fl_str_mv ark:/26339/001300000w40s
identifier_str_mv SPEROTTO, Rita Leal. Atp and adp hydrolysis in human cerebrospinal fluid. 2008. 61 f. Dissertação (Mestrado em Bioquímica) - Universidade Federal de Santa Maria, Santa Maria, 2008.
ark:/26339/001300000w40s
url http://repositorio.ufsm.br/handle/1/11088
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 de Santa Maria
BR
Bioquímica
UFSM
Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica
publisher.none.fl_str_mv Universidade Federal de Santa Maria
BR
Bioquímica
UFSM
Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica
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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