Enzima inativadora de bradicinina liberada de fígado preservado ex-vivo

Detalhes bibliográficos
Autor(a) principal: Morais, F.v. [UNIFESP]
Data de Publicação: 1999
Outros Autores: Molina, H.m. [UNIFESP], Borges, D.r. [UNIFESP], Kouyoumdjian, Maria [UNIFESP]
Tipo de documento: Artigo
Idioma: por
Título da fonte: Repositório Institucional da UNIFESP
dARK ID: ark:/48912/001300000hp65
DOI: 10.1590/S0104-42301999000100005
Texto Completo: http://dx.doi.org/10.1590/S0104-42301999000100005
http://repositorio.unifesp.br/handle/11600/747
Resumo: BACKGROUND: The liver inactivates considerable amounts of bradykinin; the main liver kinin-inactivating enzyme (BIE, bradykinin inactivating endopeptidase) hydrolyses specifically the Phe5-Ser6 bond of the nonapeptide and it has been characterized as the oligoendopeptidase E.C. 3.4.24.15. When orthotopic liver transplantation is performed there is a correlation between the increase of amino acid concentration in the preservation fluid (as a consequence of proteolysis) and graft dysfunction. AIM: Verify if BIE is released from livers stored ex vivo. METHOD: Wistar rats (180-220g) livers were exsanguinated and after removal were preserved in Braun Collins fluid or Krebs solution at 4o C. Aliquots were collected from the preservation fluid at 0, 4,8,24h, for ALT, AST, LDH and BIE assays. The fluorimetric activity of BIE was assayed upon Abz-RPPGFSPFRQ-EDDnp (synthetic BK analogue) and its presence was confirmed by immunoblotting, revealed with specific antibody anti-E.C.3.4.24.15. RESULTS: The release of ALT, AST, LDH and BIE is significant between 8-24h. In the 24h aliquots the four enzymes concentration increased in the Braun Collins fluid 8,7,19 and 10 respectively, and in the Krebs solution 21, 17, 27, 21 respectively, when compared to the zero time aliquot activities. The ratio ALT/LDH was always < 1. CONCLUSION: There is BIE release during ex vivo liver storage; this information may be useful as an indicator of the graft preservation condition; a decrease of the liver kinin-inactivating capability could affect the graft vascular reactivity.
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spelling Enzima inativadora de bradicinina liberada de fígado preservado ex-vivoBradykinin-inactivating enzyme is released from the ex-vivo stored liverBradykininEndopeptidaseCininaseLiver transplantationBradicininaEndopeptidaseFígadoCininaseTransplante hepáticoBACKGROUND: The liver inactivates considerable amounts of bradykinin; the main liver kinin-inactivating enzyme (BIE, bradykinin inactivating endopeptidase) hydrolyses specifically the Phe5-Ser6 bond of the nonapeptide and it has been characterized as the oligoendopeptidase E.C. 3.4.24.15. When orthotopic liver transplantation is performed there is a correlation between the increase of amino acid concentration in the preservation fluid (as a consequence of proteolysis) and graft dysfunction. AIM: Verify if BIE is released from livers stored ex vivo. METHOD: Wistar rats (180-220g) livers were exsanguinated and after removal were preserved in Braun Collins fluid or Krebs solution at 4o C. Aliquots were collected from the preservation fluid at 0, 4,8,24h, for ALT, AST, LDH and BIE assays. The fluorimetric activity of BIE was assayed upon Abz-RPPGFSPFRQ-EDDnp (synthetic BK analogue) and its presence was confirmed by immunoblotting, revealed with specific antibody anti-E.C.3.4.24.15. RESULTS: The release of ALT, AST, LDH and BIE is significant between 8-24h. In the 24h aliquots the four enzymes concentration increased in the Braun Collins fluid 8,7,19 and 10 respectively, and in the Krebs solution 21, 17, 27, 21 respectively, when compared to the zero time aliquot activities. The ratio ALT/LDH was always < 1. CONCLUSION: There is BIE release during ex vivo liver storage; this information may be useful as an indicator of the graft preservation condition; a decrease of the liver kinin-inactivating capability could affect the graft vascular reactivity.OBJETIVO: - O fígado inativa quantidades consideráveis de bradicinina; a principal enzima hepática cinino-inativadora (BIE, bradykinin inativating endopeptidase) hidrolisa especificamente a ligação Phe5-Ser6 do nonapeptídio e foi caracterizada como sendo a oligoendopeptidase EC 3.4. 24.15. No transplante ortotópico de fígado existe correlação entre aumento da concentração de aminoácidos no líquido de preservação (conseqüência de proteólise) e falência do enxerto. O objetivo deste trabalho é verificar se ocorre liberação da BIE de fígados preservados ex-vivo no líquido Braun-Collins ou em solução de Krebs-Henseleit bicarbonato (Krebs). MÉTODO: Fígados de ratos Wistar (180-220g) foram exsangüinados e após remoção foram preservados em líquido Braun Collins ou em solução Krebs, a 4oC. Foram retiradas alíquotas do líquido de preservação nos tempos 0, 4, 8 e 24 horas, para dosagem de ALT, AST, DHL e BIE. A atividade fluorimétrica da BIE foi ensaiada com o substrato Abz-RPPGFSPFRQ-EDDnp (análogo sintético da bradicinina) e sua presença confirmada por immunoblotting, revelado com anticorpo específico anti-EC 3.4.24.15. RESULTADOS: A liberação de ALT, AST, DHL e BIE é significativa no período 8-24hs. Nas alíquotas de 24 hs, em relação ao tempo zero, a concentração das quatro enzimas aumentou, respectivamente, no líquido Braun Collins, 8, 7, 19 e 10 vezes e, na solução de Krebs, 21, 17, 27 e 21 vezes; a relação ALT/DHL foi sempre inferior a um. CONCLUSÃO: Ocorre liberação de BIE durante a preservação ex-vivo do fígado, o que poderá servir como indicação da condição de preservação do enxerto; diminuição da capacidade cinino-inativadora do fígado poderá afetar sua reatividade vascular.Universidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Departamentos de Bioquímica e MedicinaUNIFESP, EPM, Depto.s de Bioquímica e MedicinaSciELOAssociação Médica BrasileiraUniversidade Federal de São Paulo (UNIFESP)Morais, F.v. [UNIFESP]Molina, H.m. [UNIFESP]Borges, D.r. [UNIFESP]Kouyoumdjian, Maria [UNIFESP]2015-06-14T13:24:51Z2015-06-14T13:24:51Z1999-03-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion19-23application/pdfhttp://dx.doi.org/10.1590/S0104-42301999000100005Revista da Associação Médica Brasileira. Associação Médica Brasileira, v. 45, n. 1, p. 19-23, 1999.10.1590/S0104-42301999000100005S0104-42301999000100005.pdf0104-4230S0104-42301999000100005http://repositorio.unifesp.br/handle/11600/747ark:/48912/001300000hp65porRevista da Associação Médica Brasileirainfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP2024-08-06T03:33:14Zoai:repositorio.unifesp.br/:11600/747Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestbiblioteca.csp@unifesp.bropendoar:34652024-12-11T20:19:38.695503Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false
dc.title.none.fl_str_mv Enzima inativadora de bradicinina liberada de fígado preservado ex-vivo
Bradykinin-inactivating enzyme is released from the ex-vivo stored liver
title Enzima inativadora de bradicinina liberada de fígado preservado ex-vivo
spellingShingle Enzima inativadora de bradicinina liberada de fígado preservado ex-vivo
Enzima inativadora de bradicinina liberada de fígado preservado ex-vivo
Morais, F.v. [UNIFESP]
Bradykinin
Endopeptidase
Cininase
Liver transplantation
Bradicinina
Endopeptidase
Fígado
Cininase
Transplante hepático
Morais, F.v. [UNIFESP]
Bradykinin
Endopeptidase
Cininase
Liver transplantation
Bradicinina
Endopeptidase
Fígado
Cininase
Transplante hepático
title_short Enzima inativadora de bradicinina liberada de fígado preservado ex-vivo
title_full Enzima inativadora de bradicinina liberada de fígado preservado ex-vivo
title_fullStr Enzima inativadora de bradicinina liberada de fígado preservado ex-vivo
Enzima inativadora de bradicinina liberada de fígado preservado ex-vivo
title_full_unstemmed Enzima inativadora de bradicinina liberada de fígado preservado ex-vivo
Enzima inativadora de bradicinina liberada de fígado preservado ex-vivo
title_sort Enzima inativadora de bradicinina liberada de fígado preservado ex-vivo
author Morais, F.v. [UNIFESP]
author_facet Morais, F.v. [UNIFESP]
Morais, F.v. [UNIFESP]
Molina, H.m. [UNIFESP]
Borges, D.r. [UNIFESP]
Kouyoumdjian, Maria [UNIFESP]
Molina, H.m. [UNIFESP]
Borges, D.r. [UNIFESP]
Kouyoumdjian, Maria [UNIFESP]
author_role author
author2 Molina, H.m. [UNIFESP]
Borges, D.r. [UNIFESP]
Kouyoumdjian, Maria [UNIFESP]
author2_role author
author
author
dc.contributor.none.fl_str_mv Universidade Federal de São Paulo (UNIFESP)
dc.contributor.author.fl_str_mv Morais, F.v. [UNIFESP]
Molina, H.m. [UNIFESP]
Borges, D.r. [UNIFESP]
Kouyoumdjian, Maria [UNIFESP]
dc.subject.por.fl_str_mv Bradykinin
Endopeptidase
Cininase
Liver transplantation
Bradicinina
Endopeptidase
Fígado
Cininase
Transplante hepático
topic Bradykinin
Endopeptidase
Cininase
Liver transplantation
Bradicinina
Endopeptidase
Fígado
Cininase
Transplante hepático
description BACKGROUND: The liver inactivates considerable amounts of bradykinin; the main liver kinin-inactivating enzyme (BIE, bradykinin inactivating endopeptidase) hydrolyses specifically the Phe5-Ser6 bond of the nonapeptide and it has been characterized as the oligoendopeptidase E.C. 3.4.24.15. When orthotopic liver transplantation is performed there is a correlation between the increase of amino acid concentration in the preservation fluid (as a consequence of proteolysis) and graft dysfunction. AIM: Verify if BIE is released from livers stored ex vivo. METHOD: Wistar rats (180-220g) livers were exsanguinated and after removal were preserved in Braun Collins fluid or Krebs solution at 4o C. Aliquots were collected from the preservation fluid at 0, 4,8,24h, for ALT, AST, LDH and BIE assays. The fluorimetric activity of BIE was assayed upon Abz-RPPGFSPFRQ-EDDnp (synthetic BK analogue) and its presence was confirmed by immunoblotting, revealed with specific antibody anti-E.C.3.4.24.15. RESULTS: The release of ALT, AST, LDH and BIE is significant between 8-24h. In the 24h aliquots the four enzymes concentration increased in the Braun Collins fluid 8,7,19 and 10 respectively, and in the Krebs solution 21, 17, 27, 21 respectively, when compared to the zero time aliquot activities. The ratio ALT/LDH was always < 1. CONCLUSION: There is BIE release during ex vivo liver storage; this information may be useful as an indicator of the graft preservation condition; a decrease of the liver kinin-inactivating capability could affect the graft vascular reactivity.
publishDate 1999
dc.date.none.fl_str_mv 1999-03-01
2015-06-14T13:24:51Z
2015-06-14T13:24:51Z
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1590/S0104-42301999000100005
Revista da Associação Médica Brasileira. Associação Médica Brasileira, v. 45, n. 1, p. 19-23, 1999.
10.1590/S0104-42301999000100005
S0104-42301999000100005.pdf
0104-4230
S0104-42301999000100005
http://repositorio.unifesp.br/handle/11600/747
dc.identifier.dark.fl_str_mv ark:/48912/001300000hp65
url http://dx.doi.org/10.1590/S0104-42301999000100005
http://repositorio.unifesp.br/handle/11600/747
identifier_str_mv Revista da Associação Médica Brasileira. Associação Médica Brasileira, v. 45, n. 1, p. 19-23, 1999.
10.1590/S0104-42301999000100005
S0104-42301999000100005.pdf
0104-4230
S0104-42301999000100005
ark:/48912/001300000hp65
dc.language.iso.fl_str_mv por
language por
dc.relation.none.fl_str_mv Revista da Associação Médica Brasileira
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eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 19-23
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dc.publisher.none.fl_str_mv Associação Médica Brasileira
publisher.none.fl_str_mv Associação Médica Brasileira
dc.source.none.fl_str_mv reponame:Repositório Institucional da UNIFESP
instname:Universidade Federal de São Paulo (UNIFESP)
instacron:UNIFESP
instname_str Universidade Federal de São Paulo (UNIFESP)
instacron_str UNIFESP
institution UNIFESP
reponame_str Repositório Institucional da UNIFESP
collection Repositório Institucional da UNIFESP
repository.name.fl_str_mv Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)
repository.mail.fl_str_mv biblioteca.csp@unifesp.br
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dc.identifier.doi.none.fl_str_mv 10.1590/S0104-42301999000100005