Hidrólise de atp e adp em líquor humano

Detalhes bibliográficos
Ano de defesa: 2008
Autor(a) principal: Sperotto, Rita Leal lattes
Orientador(a): Loro, Vania Lucia lattes
Banca de defesa: Schetinger, Maria Rosa Chitolina lattes, Pavanato, Maria Amália
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 Ciências Biológicas: Bioquímica Toxicológica
Departamento: Bioquímica
País: BR
Palavras-chave em Português:
Palavras-chave em Inglês:
CSF
Área do conhecimento CNPq:
Link de acesso: 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 2017-04-262017-04-262008-03-27SPEROTTO, 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/1/11088Adenine 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.application/pdfporUniversidade Federal de Santa MariaPrograma de Pós-Graduação em Ciências Biológicas: Bioquímica ToxicológicaUFSMBRBioquímicaNTPDaseCaracterizaçãoLíquido cefalorraquidianoNeuroproteçãoCSFNucleotidesNeuroprotectionCNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICAHidrólise de atp e adp em líquor humanoAtp and adp hydrolysis in human cerebrospinal fluidinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisLoro, Vania Luciahttp://lattes.cnpq.br/6392817606416780Schetinger, Maria Rosa ChitolinaPavanato, Maria Amáliahttp://lattes.cnpq.br/8701892865724171http://lattes.cnpq.br/2482919028194704Sperotto, Rita Leal200800000002400500300500500b0918185-c6b7-49df-b8d6-d245d73535b81a5493d6-e99f-48f8-8119-18bd77790024cba4a833-b1c5-4608-8453-7a46f671faf1f9c09a50-0698-4450-9e10-ac44830c8837info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALRITALEALSPEROTTO.pdfapplication/pdf559570http://repositorio.ufsm.br/bitstream/1/11088/1/RITALEALSPEROTTO.pdff72463be2b9e862e563deb553073ee58MD51TEXTRITALEALSPEROTTO.pdf.txtRITALEALSPEROTTO.pdf.txtExtracted texttext/plain76465http://repositorio.ufsm.br/bitstream/1/11088/2/RITALEALSPEROTTO.pdf.txt6f1e5ce044f16907e7b821b508eff96cMD52THUMBNAILRITALEALSPEROTTO.pdf.jpgRITALEALSPEROTTO.pdf.jpgIM Thumbnailimage/jpeg5543http://repositorio.ufsm.br/bitstream/1/11088/3/RITALEALSPEROTTO.pdf.jpg07901d2ceebf5f3c6e1b06aaf72eeedcMD531/110882022-08-25 16:35:28.949oai:repositorio.ufsm.br:1/11088Repositório Institucionalhttp://repositorio.ufsm.br/PUBhttp://repositorio.ufsm.br/oai/requestopendoar:39132022-08-25T19:35:28Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.por.fl_str_mv Hidrólise de atp e adp em líquor humano
dc.title.alternative.eng.fl_str_mv 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.advisor1.fl_str_mv Loro, Vania Lucia
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/6392817606416780
dc.contributor.referee1.fl_str_mv Schetinger, Maria Rosa Chitolina
dc.contributor.referee2.fl_str_mv Pavanato, Maria Amália
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/8701892865724171
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/2482919028194704
dc.contributor.author.fl_str_mv Sperotto, Rita Leal
contributor_str_mv Loro, Vania Lucia
Schetinger, Maria Rosa Chitolina
Pavanato, Maria Amália
dc.subject.por.fl_str_mv NTPDase
Caracterização
Líquido cefalorraquidiano
Neuroproteção
topic NTPDase
Caracterização
Líquido cefalorraquidiano
Neuroproteção
CSF
Nucleotides
Neuroprotection
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
dc.subject.eng.fl_str_mv CSF
Nucleotides
Neuroprotection
dc.subject.cnpq.fl_str_mv 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.issued.fl_str_mv 2008-03-27
dc.date.accessioned.fl_str_mv 2017-04-26
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dc.identifier.citation.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.
dc.identifier.uri.fl_str_mv http://repositorio.ufsm.br/handle/1/11088
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.
url http://repositorio.ufsm.br/handle/1/11088
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