Hidrólise de atp e adp em líquor humano
Ano de defesa: | 2008 |
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Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | , |
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: | |
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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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 |
dc.date.available.fl_str_mv |
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.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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Universidade Federal de Santa Maria |
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Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica |
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