Modulação da proliferação, apoptose e autofagia de linfocitos T por RAGE
Ano de defesa: | 2014 |
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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 Estadual Paulista (Unesp)
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Programa de Pós-Graduação: |
Não Informado pela instituição
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Departamento: |
Não Informado pela instituição
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País: |
Não Informado pela instituição
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Palavras-chave em Português: | |
Link de acesso: | http://hdl.handle.net/11449/110823 |
Resumo: | RAGE (Receptor for advanced glycation endproducts) is a pattern recognition receptor (PRR) that is especially recognized for its involvement in the pathogenesis of secondary complications of Diabetes Mellitus. Its activation is related to the modulation of immune-inflammatory responses, by promoting activation, maturation and migration of leukocytes, as well as the induction of inflammatory cytokines. RAGE signaling is also related to the regulation of cellular processes like survival, apoptosis and autophagy. Little is known about the role of RAGE in the regulation of these processes in cells of the adaptive immune response, specifically in T lymphocytes. Our main purpose was to evaluate the influence of RAGE activation in T cell biology. We observed the role of this receptor in the modulation of proliferation, apoptosis and autophagy, and the contribution of NF-kB in the induction of these processes. A human cell line of T lymphocytes (JM/ Jurkat) were stimulated with the RAGE ligands: BSA-AGE (100 and 200 μg/mL) and S100B (10 μg/mL). The stimulation was also performed after pre-activation of the T cell receptor (TCR). Gain of function experiments were performed by transient transfection of cells with a plasmid vector to increase RAGE expression. Cell proliferation was determined by Trypan blue dye exclusion assay. Apoptosis was assessed by the detection of DNA fragmentation by the TUNEL method, and p53, Bax and Bcl-2 expression by Western blotting. Also by Western blotting, we observed the effect of RAGE on the expression of autophagy-related proteins, p62 and LC3, and on the expression of p50, subunit of the transcription factor, NF-κB. The stimulation with BSA- AGE and S100b did not alter the proliferation and viability, but, in transfected cells, this stimulation tended to increase proliferation and viability, especially after the cell pre-activation. BSA-AGE and S100b had distinct effects on apoptosis depending on the... |
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Modulação da proliferação, apoptose e autofagia de linfocitos T por RAGECelulas TProdutos finais de glicosilaçãoPeriodontiaPeriodonticsRAGE (Receptor for advanced glycation endproducts) is a pattern recognition receptor (PRR) that is especially recognized for its involvement in the pathogenesis of secondary complications of Diabetes Mellitus. Its activation is related to the modulation of immune-inflammatory responses, by promoting activation, maturation and migration of leukocytes, as well as the induction of inflammatory cytokines. RAGE signaling is also related to the regulation of cellular processes like survival, apoptosis and autophagy. Little is known about the role of RAGE in the regulation of these processes in cells of the adaptive immune response, specifically in T lymphocytes. Our main purpose was to evaluate the influence of RAGE activation in T cell biology. We observed the role of this receptor in the modulation of proliferation, apoptosis and autophagy, and the contribution of NF-kB in the induction of these processes. A human cell line of T lymphocytes (JM/ Jurkat) were stimulated with the RAGE ligands: BSA-AGE (100 and 200 μg/mL) and S100B (10 μg/mL). The stimulation was also performed after pre-activation of the T cell receptor (TCR). Gain of function experiments were performed by transient transfection of cells with a plasmid vector to increase RAGE expression. Cell proliferation was determined by Trypan blue dye exclusion assay. Apoptosis was assessed by the detection of DNA fragmentation by the TUNEL method, and p53, Bax and Bcl-2 expression by Western blotting. Also by Western blotting, we observed the effect of RAGE on the expression of autophagy-related proteins, p62 and LC3, and on the expression of p50, subunit of the transcription factor, NF-κB. The stimulation with BSA- AGE and S100b did not alter the proliferation and viability, but, in transfected cells, this stimulation tended to increase proliferation and viability, especially after the cell pre-activation. BSA-AGE and S100b had distinct effects on apoptosis depending on the...moleculares (PRR) reconhecido principalmente por seu envolvimento na patogênese de complicações secundárias do Diabetes Mellitus. Sua ativação está relacionada à modulação da resposta imune-inflamatória, promovendo ativação, migração e maturação de leucócitos; e induzindo a liberação de citocinas inflamatórias. A sinalização via RAGE também está relacionada à regulação dos processos de sobrevivência, apoptose e autofagia celular. Pouco se sabe sobre o papel de RAGE na regulação desses processos em células da resposta imune adaptativa e, especificamente, em linfócitos T. Nosso objetivo foi avaliar a influência da ativação de RAGE na biologia de células T. Para isso, observamos seus efeitos na modulação da proliferação, apoptose e autofagia, e a contribuição da ativação de NF-κB na indução desses processos. Células de linhagem de linfócitos T (JM/Jurkat) foram estimuladas com os ligantes de RAGE BSA-AGE (100 e 200 μg/mL) e S100b (10 μg/mL). A estimulação também foi realizada mediante a pré-ativação do receptor de células T (TCR). Experimentos de ganho de função foram realizados por meio da transfecção transitória das células com um vetor plasmidial de RAGE. A proliferação celular foi determinada por ensaio de exclusão com azul de Trypan. A apoptose foi avaliada através da detecção da fragmentação do DNA pelo método TUNEL e da expressão de p53, Bax e Bcl-2 por Western blot. Ainda por Western blot, observamos a expressão de proteínas características do processo de autofagia, p62 e LC3; e de p50, subunidade do fator de transcrição NF-κB. O estímulo com BSA-AGE e S100b não alterou a proliferação e a viabilidade, mas, em células transfectadas, esse estímulo tendeu a aumentar a proliferação e a viabilidade, sobretudo após a pré-ativação das células. O BSA-AGE e o S100b produziram efeitos distintos sobre a apoptose a depender da pré-ativação e do período...Universidade Estadual Paulista (Unesp)Rossa Júnior, Carlos [UNESP]Universidade Estadual Paulista (Unesp)Barbosa, Ligia Araujo [UNESP]2014-12-02T11:16:37Z2014-12-02T11:16:37Z2014-03-21info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis94 f. : il. color. + anexoapplication/pdfBARBOSA, Ligia Araujo. Modulação da proliferação, apoptose e autofagia de linfocitos T por RAGE. 2014. 94 f. Dissertação(mestrado) - Universidade Estadual Paulista Julio de Mesquita Filho, Faculdade de Odontologia de Araraquara, 2014.http://hdl.handle.net/11449/110823000797900000797900.pdf33004030059P1Alephreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPporinfo:eu-repo/semantics/openAccess2024-09-30T14:34:26Zoai:repositorio.unesp.br:11449/110823Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestrepositoriounesp@unesp.bropendoar:29462024-09-30T14:34:26Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Modulação da proliferação, apoptose e autofagia de linfocitos T por RAGE |
title |
Modulação da proliferação, apoptose e autofagia de linfocitos T por RAGE |
spellingShingle |
Modulação da proliferação, apoptose e autofagia de linfocitos T por RAGE Barbosa, Ligia Araujo [UNESP] Celulas T Produtos finais de glicosilação Periodontia Periodontics |
title_short |
Modulação da proliferação, apoptose e autofagia de linfocitos T por RAGE |
title_full |
Modulação da proliferação, apoptose e autofagia de linfocitos T por RAGE |
title_fullStr |
Modulação da proliferação, apoptose e autofagia de linfocitos T por RAGE |
title_full_unstemmed |
Modulação da proliferação, apoptose e autofagia de linfocitos T por RAGE |
title_sort |
Modulação da proliferação, apoptose e autofagia de linfocitos T por RAGE |
author |
Barbosa, Ligia Araujo [UNESP] |
author_facet |
Barbosa, Ligia Araujo [UNESP] |
author_role |
author |
dc.contributor.none.fl_str_mv |
Rossa Júnior, Carlos [UNESP] Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Barbosa, Ligia Araujo [UNESP] |
dc.subject.por.fl_str_mv |
Celulas T Produtos finais de glicosilação Periodontia Periodontics |
topic |
Celulas T Produtos finais de glicosilação Periodontia Periodontics |
description |
RAGE (Receptor for advanced glycation endproducts) is a pattern recognition receptor (PRR) that is especially recognized for its involvement in the pathogenesis of secondary complications of Diabetes Mellitus. Its activation is related to the modulation of immune-inflammatory responses, by promoting activation, maturation and migration of leukocytes, as well as the induction of inflammatory cytokines. RAGE signaling is also related to the regulation of cellular processes like survival, apoptosis and autophagy. Little is known about the role of RAGE in the regulation of these processes in cells of the adaptive immune response, specifically in T lymphocytes. Our main purpose was to evaluate the influence of RAGE activation in T cell biology. We observed the role of this receptor in the modulation of proliferation, apoptosis and autophagy, and the contribution of NF-kB in the induction of these processes. A human cell line of T lymphocytes (JM/ Jurkat) were stimulated with the RAGE ligands: BSA-AGE (100 and 200 μg/mL) and S100B (10 μg/mL). The stimulation was also performed after pre-activation of the T cell receptor (TCR). Gain of function experiments were performed by transient transfection of cells with a plasmid vector to increase RAGE expression. Cell proliferation was determined by Trypan blue dye exclusion assay. Apoptosis was assessed by the detection of DNA fragmentation by the TUNEL method, and p53, Bax and Bcl-2 expression by Western blotting. Also by Western blotting, we observed the effect of RAGE on the expression of autophagy-related proteins, p62 and LC3, and on the expression of p50, subunit of the transcription factor, NF-κB. The stimulation with BSA- AGE and S100b did not alter the proliferation and viability, but, in transfected cells, this stimulation tended to increase proliferation and viability, especially after the cell pre-activation. BSA-AGE and S100b had distinct effects on apoptosis depending on the... |
publishDate |
2014 |
dc.date.none.fl_str_mv |
2014-12-02T11:16:37Z 2014-12-02T11:16:37Z 2014-03-21 |
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 |
BARBOSA, Ligia Araujo. Modulação da proliferação, apoptose e autofagia de linfocitos T por RAGE. 2014. 94 f. Dissertação(mestrado) - Universidade Estadual Paulista Julio de Mesquita Filho, Faculdade de Odontologia de Araraquara, 2014. http://hdl.handle.net/11449/110823 000797900 000797900.pdf 33004030059P1 |
identifier_str_mv |
BARBOSA, Ligia Araujo. Modulação da proliferação, apoptose e autofagia de linfocitos T por RAGE. 2014. 94 f. Dissertação(mestrado) - Universidade Estadual Paulista Julio de Mesquita Filho, Faculdade de Odontologia de Araraquara, 2014. 000797900 000797900.pdf 33004030059P1 |
url |
http://hdl.handle.net/11449/110823 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
94 f. : il. color. + anexo application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
publisher.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
dc.source.none.fl_str_mv |
Aleph reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
repositoriounesp@unesp.br |
_version_ |
1813546880142934016 |