Enzima inativadora de bradicinina liberada de fígado preservado ex-vivo
Autor(a) principal: | |
---|---|
Data de Publicação: | 1999 |
Outros Autores: | , , |
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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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 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
19-23 application/pdf |
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 |
_version_ |
1822183937663827968 |
dc.identifier.doi.none.fl_str_mv |
10.1590/S0104-42301999000100005 |