Reactivity of human fetal and adult immortalized hepatocytes to potentially toxic bilirubin and bile acid species

Detalhes bibliográficos
Autor(a) principal: Duarte, Catarina Vizetto Guerreiro
Data de Publicação: 2009
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/2624
Resumo: Dissertação apresentada para a obtenção do Grau de Mestre em Genética Molecular e Biomedicina, pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia
id RCAP_fda51b5159689616e3366e19c3d3a19b
oai_identifier_str oai:run.unl.pt:10362/2624
network_acronym_str RCAP
network_name_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository_id_str 7160
spelling Reactivity of human fetal and adult immortalized hepatocytes to potentially toxic bilirubin and bile acid speciesDissertação apresentada para a obtenção do Grau de Mestre em Genética Molecular e Biomedicina, pela Universidade Nova de Lisboa, Faculdade de Ciências e TecnologiaCholestasis is the reduction or stoppage of bile flow. When bile flow is interrupted, the bile compounds, namely bile acids and bilirubin, accumulate into the hepatocyte causing cellular injury and cell death. The hepatocyte function under cholestasis must be studied using liver cell lines closely resembling human primary hepatocytes. However, most of the cell lines used is derived from hepatic tumors which have altered gene expression. In this project, we used the novel non-neoplasic cell line, HHL-5, which retain primary adult hepatocyte phenotype. In addition, we used the fetal hepatocyte cell line WRL-68, which was shown to present similar morphological properties and antigenic profile of human fetal hepatocytes in situ. Using these two cell lines, we evaluated whether fetal and adult hepatocytes respond differently to conditions that mimic cholestasis with associated jaundice, never evaluated in vitro before. The hepatocytes were exposed to 100 μM glycochenodeoxycholic acid(GCDCA), 100 μM conjugated bilirubin (CB), 100 μM unconjugated bilirubin (UCB), 100 μM GCDCA + 100 μM CB + 100 μM UCB or vehicle alone, in the presence of 100 μM human serum albumin (HSA), at various time points. After, we assessed cellular toxicity analyzing cytolysis by lactate dehydrogenase (LDH) release and apoptosis by caspase-3 activity and nuclear fragmentation. There was a significant increase of all these parameters in hepatocytes stimulated with the GCDCA+CB+UCB mixture. LDH release significantly increased after 48 h incubation for GCDCA+CB+UCB treatment (~40%, P<0.01) in the adult cell line. This increase was also statistically significant when compared to GCDCA, CB or UCB incubation alone (P<0.05). Relatively to WRL-68 cells, the release of LDH also increased significantly (~22%) after 48 h incubation with GCDCA+CB+UCB when compared to control (P<0.05), GCDCA (P<0.05) or UCB (P<0.01) incubations. Regarding caspase-3 activity, the increase was evident for GCDCA+CB+UCB treatment. In the adult cell line, it increased ~1.5-fold after 6 h (P<0.01 vs. control, P<0.01 vs. GCDCA, P<0.05 vs. UCB), peaking at 12 h (~3.5-fold, P<0.05 vs. control, P<0.05 vs. GCDCA) and remaining elevated after 24 h (P<0.01 vs. control, P<0.01 vs. GCDCA, P<0.05 vs. CB, P<0.01 vs. UCB). In the fetal cell line, the peak of caspase-3 activity with the same treatment occurred earlier and in a higher magnitude. Indeed, caspase-3 activity increased 4-fold after GCDCA+CB+UCB treatment at 6 h incubation (P<0.05) and this activation was sustained until 12 h incubation(P<0.01). Concerning nuclear fragmentation, in both cell lines, hepatocytes incubated with GCDCA+CB+UCB exhibited profound changes in nuclear morphology, consistent with apoptosis, in a more marked way than when incubated with GCDCA, CB or UCB alone. The results showed similar data to those previously obtained for caspase-3 activity.FCT - UNLBrites, DoraFernandes, AdelaideRUNDuarte, Catarina Vizetto Guerreiro2010-02-08T14:45:55Z20092009-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/2624enginfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T03:32:52Zoai:run.unl.pt:10362/2624Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:15:14.279219Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Reactivity of human fetal and adult immortalized hepatocytes to potentially toxic bilirubin and bile acid species
title Reactivity of human fetal and adult immortalized hepatocytes to potentially toxic bilirubin and bile acid species
spellingShingle Reactivity of human fetal and adult immortalized hepatocytes to potentially toxic bilirubin and bile acid species
Duarte, Catarina Vizetto Guerreiro
title_short Reactivity of human fetal and adult immortalized hepatocytes to potentially toxic bilirubin and bile acid species
title_full Reactivity of human fetal and adult immortalized hepatocytes to potentially toxic bilirubin and bile acid species
title_fullStr Reactivity of human fetal and adult immortalized hepatocytes to potentially toxic bilirubin and bile acid species
title_full_unstemmed Reactivity of human fetal and adult immortalized hepatocytes to potentially toxic bilirubin and bile acid species
title_sort Reactivity of human fetal and adult immortalized hepatocytes to potentially toxic bilirubin and bile acid species
author Duarte, Catarina Vizetto Guerreiro
author_facet Duarte, Catarina Vizetto Guerreiro
author_role author
dc.contributor.none.fl_str_mv Brites, Dora
Fernandes, Adelaide
RUN
dc.contributor.author.fl_str_mv Duarte, Catarina Vizetto Guerreiro
description Dissertação apresentada para a obtenção do Grau de Mestre em Genética Molecular e Biomedicina, pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia
publishDate 2009
dc.date.none.fl_str_mv 2009
2009-01-01T00:00:00Z
2010-02-08T14:45:55Z
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 http://hdl.handle.net/10362/2624
url http://hdl.handle.net/10362/2624
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv FCT - UNL
publisher.none.fl_str_mv FCT - UNL
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv
_version_ 1799137803824529408

Registros relacionados