Estudo morfofuncional e dos mediadores inflamatórios envolvidos na patogênese da mucosite intestinal induzida por irinotecano (CPT-11) em camundongos : papel da caspase-1, interleucina-18 e óxido nitríco
Autor(a) principal: | |
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Data de Publicação: | 2008 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da Universidade Federal do Ceará (UFC) |
dARK ID: | ark:/83112/001300000z1c8 |
Texto Completo: | http://www.repositorio.ufc.br/handle/riufc/3843 |
Resumo: | Severe Intestinal Mucositis (IM) and diarrhea are frequent and dose-limiting side-effects of colon cancer chemotherapy with irinotecan (CPT-11). At present, much of IM pathophysiology remains unknown. Aims: To study the mechanisms and inflammatory mediators involved in CPT-11-induced intestinal mucositis, verifying the role of the protease caspase-1, cytokines interleukin-1 (IL-1), interleukin-18 (IL-18) and interleukin-33 (IL-33), nitric oxide, 5-lipoxygenase and PAF as well as the sequence of activation of these mediators in the inflammatory response and morphofunctional contexts. Methods: Swiss, C57BL/6 (BL), BALB/c (BC) male mice or caspases-1(Casp-1-/-), IL-18 (IL-18-/-), inducible nitric oxide synthase (iNOS-/-), 5-Lipoxygenase (5-LOX-/-), PAF receptor (PAFr-/-) knockout mice, 22g, were divided into groups (n=4-5) and treated for 4 days with saline (5 mL/kg, i.p.) or CPT-11 (60 mg/kg, i.p.) or were given IL-18bp (IL-18 binding protein, 200 µg/animal/4 days, i.p.), aminoguanidine (AG, 50 mg/kg, s.c, 2x/day/4 days), IL-33 (1 µg/animal/4 days, i.v, 1h previously CPT-11) or loperamide ([4x3 mg/kg and 4x30 mg/kg]/animal, s.c.) co-administered with CPT-11. On day 5, diarrhea and leukocyte counts were assessed, and following sacrifice duodenal portions of the animal were collected to assess myeloperoxidase (MPO, neutrophils/mg tissue) and nitric oxide synthase (iNOS pM citruline /h/mg protein) activity, morphometric analysis, and in vitro contractility (%contraction in relation to KCl 60 mM). ANOVA/Newman Keuls or Kruskal Wallis/Dunn were used as statistical tests. P<0.05 was accepted. Results: IM was morphofunctionally similar in both BL and Swiss. CPT-11 induced leukopenia in all animal lineages despite the treatment given. Additionally, CPT-11 induced MPO, increased in vitro contractility and diarrheic events, and villi/crypt ratio reduction in BL and BC mice in comparison with saline treated mice (p<0.05). Despite the injection of CPT-11, Casp-1-/-, IL-18-/- or IL-18bp treated BC mice presented reduced MPO and iNOS activities and also increased villi/crypt ratio, reduced duodenal in vitro contractility and mild diarrhea similar to saline-treated mice (P>0.05), similar to the patterns seen in IL-33 treated mice. The reduced iNOS activity found in the earlier mice led us to investigate the CPT-11 effect on iNOS /- and aminoguanidine-treated mice. Lower IL-1beta and KC cytokine levels, as well as reduced MPO activity were found. Also, preserved villi/crypt ratio and muscle layer thickness, normal duodenal contractility and diarrheic events were seen similarly to saline-treated mice (p>0.05). However, CPT-11-treated PAFr-/- presented morphofunctional alterations and increased intestinal MPO activity. 5-LOX-/- or loperamide treated mice (positive control), both injected with CPT-11, exhibited increased MPO activity and morphometric alterations, but no exacerbation on in vitro contractility was observable. Conclusion: Taken together, these results suggest the role of caspase-1/Interleukin-18/nitric oxide cascade on pathogenesis of CPT-11-induced morphofunctional alteration and inflammatory response (IM). IL-33 seems to be a compensatory protective factor on this inflammatory cascade. 5-LOX and PAFr show a likely secondary role on IM pathophysiology. |
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Estudo morfofuncional e dos mediadores inflamatórios envolvidos na patogênese da mucosite intestinal induzida por irinotecano (CPT-11) em camundongos : papel da caspase-1, interleucina-18 e óxido nitrícoMorphofunctional study and the inflammatory mediators involved on the pathogenesis of irinotecan (CPT-11)-induced intestinal mucositis in mice : role of caspase-1, interleukin-18 and nitric oxideMucositeFator de Ativação de PlaquetasMediadores da InflamaçãoInflamaçãoCitocinasSevere Intestinal Mucositis (IM) and diarrhea are frequent and dose-limiting side-effects of colon cancer chemotherapy with irinotecan (CPT-11). At present, much of IM pathophysiology remains unknown. Aims: To study the mechanisms and inflammatory mediators involved in CPT-11-induced intestinal mucositis, verifying the role of the protease caspase-1, cytokines interleukin-1 (IL-1), interleukin-18 (IL-18) and interleukin-33 (IL-33), nitric oxide, 5-lipoxygenase and PAF as well as the sequence of activation of these mediators in the inflammatory response and morphofunctional contexts. Methods: Swiss, C57BL/6 (BL), BALB/c (BC) male mice or caspases-1(Casp-1-/-), IL-18 (IL-18-/-), inducible nitric oxide synthase (iNOS-/-), 5-Lipoxygenase (5-LOX-/-), PAF receptor (PAFr-/-) knockout mice, 22g, were divided into groups (n=4-5) and treated for 4 days with saline (5 mL/kg, i.p.) or CPT-11 (60 mg/kg, i.p.) or were given IL-18bp (IL-18 binding protein, 200 µg/animal/4 days, i.p.), aminoguanidine (AG, 50 mg/kg, s.c, 2x/day/4 days), IL-33 (1 µg/animal/4 days, i.v, 1h previously CPT-11) or loperamide ([4x3 mg/kg and 4x30 mg/kg]/animal, s.c.) co-administered with CPT-11. On day 5, diarrhea and leukocyte counts were assessed, and following sacrifice duodenal portions of the animal were collected to assess myeloperoxidase (MPO, neutrophils/mg tissue) and nitric oxide synthase (iNOS pM citruline /h/mg protein) activity, morphometric analysis, and in vitro contractility (%contraction in relation to KCl 60 mM). ANOVA/Newman Keuls or Kruskal Wallis/Dunn were used as statistical tests. P<0.05 was accepted. Results: IM was morphofunctionally similar in both BL and Swiss. CPT-11 induced leukopenia in all animal lineages despite the treatment given. Additionally, CPT-11 induced MPO, increased in vitro contractility and diarrheic events, and villi/crypt ratio reduction in BL and BC mice in comparison with saline treated mice (p<0.05). Despite the injection of CPT-11, Casp-1-/-, IL-18-/- or IL-18bp treated BC mice presented reduced MPO and iNOS activities and also increased villi/crypt ratio, reduced duodenal in vitro contractility and mild diarrhea similar to saline-treated mice (P>0.05), similar to the patterns seen in IL-33 treated mice. The reduced iNOS activity found in the earlier mice led us to investigate the CPT-11 effect on iNOS /- and aminoguanidine-treated mice. Lower IL-1beta and KC cytokine levels, as well as reduced MPO activity were found. Also, preserved villi/crypt ratio and muscle layer thickness, normal duodenal contractility and diarrheic events were seen similarly to saline-treated mice (p>0.05). However, CPT-11-treated PAFr-/- presented morphofunctional alterations and increased intestinal MPO activity. 5-LOX-/- or loperamide treated mice (positive control), both injected with CPT-11, exhibited increased MPO activity and morphometric alterations, but no exacerbation on in vitro contractility was observable. Conclusion: Taken together, these results suggest the role of caspase-1/Interleukin-18/nitric oxide cascade on pathogenesis of CPT-11-induced morphofunctional alteration and inflammatory response (IM). IL-33 seems to be a compensatory protective factor on this inflammatory cascade. 5-LOX and PAFr show a likely secondary role on IM pathophysiology.A Mucosite Intestinal (MI) e a diarréia severas são efeitos colaterais freqüentes (15-25%) e limitantes da quimioterapia do câncer de cólon com Irinotecano (CPT-11). Até o presente momento, pouco se conhece sobre a fisiopatologia da MI. Objetivo: Estudar os mecanismos e mediadores envolvidos na mucosite intestinal induzida por CPT-11, verificando a participação da protease caspase-1, das citocinas interleucina-1 (IL-1), interleucina-18 (IL-18) e interleucina-33 (IL-33), do óxido nítrico, de mediadores 5-lipoxigenase e do PAF, assim como a seqüência de interação entre esses mediadores sob aspectos da resposta inflamatória e morfofuncionais. Métodos: Camundongos Swiss, C57BL/6 (BL), BALB/c (BC) ou knockout para Caspase-1 (Casp-/-), IL-18 (IL-18-/-), óxido nítrico sintase induzida (iNOS-/-), 5-Lipoxigenase (5-LOX-/-), receptor para PAF (PAFr-/-) machos, 22 g, foram divididos em grupos (n=4-5) e tratados por 4 dias com salina (5 mL/kg, i.p) ou CPT-11 (60 mg/kg, i.p) ou foram pré-tratados com IL-18bp (proteína ligante de IL18, 200 µg/animal/4 dias, i.p.), aminoguanidina (AG, 50 mg/kg, s.c, 2x/dia/4 dias), IL-33 (1 µg/animal/4 dias, i.v, 1h antes do CPT-11) ou loperamida ([4x3 mg/kg e 4x30 mg/kg]/animal, s.c.) em associação com CPT-11. No 5º dia, avaliou-se a diarréia por escores, o leucograma e, após sacrifício, coletou-se o duodeno para dosagem da atividade de mieloperoxidase (MPO, neutrófilos/mg tecido) e de óxido nítrico sintase induzida (iNOS pM citrulina/h/mg proteína), morfometria (razão vilo/cripta) e contratilidade in vitro à Acetilcolina (%contração em relação ao KCl60 mM). Para análise estatística utilizou-se ANOVA/Newman-Keuls ou Kruskal Wallis/Dunn. P<0,05 foi aceito. Resultados: Camundongos BL apresentaram um padrão de mucosite intestinal similar ao observado no camundongo Swiss sob aspectos morfofuncionais. O CPT-11 induziu leucopenia em todos os animais independentemente do tratamento aplicado. Adicionalmente, o CPT-11 induziu em animais BL e BC, não tratados, aumento de MPO intestinal, redução da relação vilo/cripta, amento da contratilidade in vitro e dos eventos de diarréia comparados com animais injetados somente com salina (p<0,05). A despeito da administração de CPT-11, animais Caspase-1-/-, IL-18-/- ou BC tratados com IL-18bp apresentaram atividade de MPO e de iNOS reduzidas, bem como aumento na relação vilo/cripta, menor contratilidade duodenal in vitro e eventos de diarréia atenuados em comparação aos respectivos controles tratados com salina (p>0,05). Dados idênticos também foram observados em camundongos injetados com IL-33 e que receberam CPT-11. A redução na atividade de iNOS naqueles animais motivou o estudo do efeito do CPT-11 em animais iNOS-/- ou tratados com aminoguanidina. Observaram-se menores níveis de citocinas IL-1beta e KC, bem como uma atividade de MPO reduzida no duodeno desses animais. Além disso, verificou-se preservação da relação vilo/cripta e da espessura da camada muscular, menor contratilidade duodenal in vitro e eventos de diarréia atenuados similar ao observado nos respectivos controles que receberam apenas salina (p>0,05). Porém, a administração de CPT-11 a animais PAFr-/- foi capaz de induzir alterações morfofuncionais e o aumento da atividade de MPO intestinal. Em camundongos 5-LOX /- ou tratados com loperamida (controle positivo), ambos os grupos injetados com CPT-11, observaram-se aumento da atividade de MPO e alterações morfométricas. Contudo, sobre aspectos funcionais de contratilidade in vitro, visualizou-se proteção. Conclusões: Esses resultados sugerem que a via caspase-1-IL-18 óxido nítrico contribui para o desenvolvimento da resposta inflamatória e alterações morfofuncionais na mucosite intestinal induzida por CPT-11. A IL-33 parece ser um fator protetor compensatório nessa cascata inflamatória. A 5-LOX e o PAFr parecem ter papel secundário na patogênese da mucosite intestinal por CPT-11.Ribeiro, Ronaldo de AlbuquerqueLima Júnior, Roberto César Pereira2012-10-01T16:30:37Z2012-10-01T16:30:37Z2008info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfLIMA JUNIOR, R. C. P. Estudo morfofuncional e dos mediadores inflamatórios envolvidos na patogênese da mucosite intestinal induzida por irinotecano (CPT-11) em camundongos : papel da caspase-1, interleucina-18 e óxido nitríco. 200 f. Tese (Doutorado em Farmacologia) - Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2008.http://www.repositorio.ufc.br/handle/riufc/3843ark:/83112/001300000z1c8porreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccess2021-06-29T13:54:11Zoai:repositorio.ufc.br:riufc/3843Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-09-11T18:20:22.767153Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false |
dc.title.none.fl_str_mv |
Estudo morfofuncional e dos mediadores inflamatórios envolvidos na patogênese da mucosite intestinal induzida por irinotecano (CPT-11) em camundongos : papel da caspase-1, interleucina-18 e óxido nitríco Morphofunctional study and the inflammatory mediators involved on the pathogenesis of irinotecan (CPT-11)-induced intestinal mucositis in mice : role of caspase-1, interleukin-18 and nitric oxide |
title |
Estudo morfofuncional e dos mediadores inflamatórios envolvidos na patogênese da mucosite intestinal induzida por irinotecano (CPT-11) em camundongos : papel da caspase-1, interleucina-18 e óxido nitríco |
spellingShingle |
Estudo morfofuncional e dos mediadores inflamatórios envolvidos na patogênese da mucosite intestinal induzida por irinotecano (CPT-11) em camundongos : papel da caspase-1, interleucina-18 e óxido nitríco Lima Júnior, Roberto César Pereira Mucosite Fator de Ativação de Plaquetas Mediadores da Inflamação Inflamação Citocinas |
title_short |
Estudo morfofuncional e dos mediadores inflamatórios envolvidos na patogênese da mucosite intestinal induzida por irinotecano (CPT-11) em camundongos : papel da caspase-1, interleucina-18 e óxido nitríco |
title_full |
Estudo morfofuncional e dos mediadores inflamatórios envolvidos na patogênese da mucosite intestinal induzida por irinotecano (CPT-11) em camundongos : papel da caspase-1, interleucina-18 e óxido nitríco |
title_fullStr |
Estudo morfofuncional e dos mediadores inflamatórios envolvidos na patogênese da mucosite intestinal induzida por irinotecano (CPT-11) em camundongos : papel da caspase-1, interleucina-18 e óxido nitríco |
title_full_unstemmed |
Estudo morfofuncional e dos mediadores inflamatórios envolvidos na patogênese da mucosite intestinal induzida por irinotecano (CPT-11) em camundongos : papel da caspase-1, interleucina-18 e óxido nitríco |
title_sort |
Estudo morfofuncional e dos mediadores inflamatórios envolvidos na patogênese da mucosite intestinal induzida por irinotecano (CPT-11) em camundongos : papel da caspase-1, interleucina-18 e óxido nitríco |
author |
Lima Júnior, Roberto César Pereira |
author_facet |
Lima Júnior, Roberto César Pereira |
author_role |
author |
dc.contributor.none.fl_str_mv |
Ribeiro, Ronaldo de Albuquerque |
dc.contributor.author.fl_str_mv |
Lima Júnior, Roberto César Pereira |
dc.subject.por.fl_str_mv |
Mucosite Fator de Ativação de Plaquetas Mediadores da Inflamação Inflamação Citocinas |
topic |
Mucosite Fator de Ativação de Plaquetas Mediadores da Inflamação Inflamação Citocinas |
description |
Severe Intestinal Mucositis (IM) and diarrhea are frequent and dose-limiting side-effects of colon cancer chemotherapy with irinotecan (CPT-11). At present, much of IM pathophysiology remains unknown. Aims: To study the mechanisms and inflammatory mediators involved in CPT-11-induced intestinal mucositis, verifying the role of the protease caspase-1, cytokines interleukin-1 (IL-1), interleukin-18 (IL-18) and interleukin-33 (IL-33), nitric oxide, 5-lipoxygenase and PAF as well as the sequence of activation of these mediators in the inflammatory response and morphofunctional contexts. Methods: Swiss, C57BL/6 (BL), BALB/c (BC) male mice or caspases-1(Casp-1-/-), IL-18 (IL-18-/-), inducible nitric oxide synthase (iNOS-/-), 5-Lipoxygenase (5-LOX-/-), PAF receptor (PAFr-/-) knockout mice, 22g, were divided into groups (n=4-5) and treated for 4 days with saline (5 mL/kg, i.p.) or CPT-11 (60 mg/kg, i.p.) or were given IL-18bp (IL-18 binding protein, 200 µg/animal/4 days, i.p.), aminoguanidine (AG, 50 mg/kg, s.c, 2x/day/4 days), IL-33 (1 µg/animal/4 days, i.v, 1h previously CPT-11) or loperamide ([4x3 mg/kg and 4x30 mg/kg]/animal, s.c.) co-administered with CPT-11. On day 5, diarrhea and leukocyte counts were assessed, and following sacrifice duodenal portions of the animal were collected to assess myeloperoxidase (MPO, neutrophils/mg tissue) and nitric oxide synthase (iNOS pM citruline /h/mg protein) activity, morphometric analysis, and in vitro contractility (%contraction in relation to KCl 60 mM). ANOVA/Newman Keuls or Kruskal Wallis/Dunn were used as statistical tests. P<0.05 was accepted. Results: IM was morphofunctionally similar in both BL and Swiss. CPT-11 induced leukopenia in all animal lineages despite the treatment given. Additionally, CPT-11 induced MPO, increased in vitro contractility and diarrheic events, and villi/crypt ratio reduction in BL and BC mice in comparison with saline treated mice (p<0.05). Despite the injection of CPT-11, Casp-1-/-, IL-18-/- or IL-18bp treated BC mice presented reduced MPO and iNOS activities and also increased villi/crypt ratio, reduced duodenal in vitro contractility and mild diarrhea similar to saline-treated mice (P>0.05), similar to the patterns seen in IL-33 treated mice. The reduced iNOS activity found in the earlier mice led us to investigate the CPT-11 effect on iNOS /- and aminoguanidine-treated mice. Lower IL-1beta and KC cytokine levels, as well as reduced MPO activity were found. Also, preserved villi/crypt ratio and muscle layer thickness, normal duodenal contractility and diarrheic events were seen similarly to saline-treated mice (p>0.05). However, CPT-11-treated PAFr-/- presented morphofunctional alterations and increased intestinal MPO activity. 5-LOX-/- or loperamide treated mice (positive control), both injected with CPT-11, exhibited increased MPO activity and morphometric alterations, but no exacerbation on in vitro contractility was observable. Conclusion: Taken together, these results suggest the role of caspase-1/Interleukin-18/nitric oxide cascade on pathogenesis of CPT-11-induced morphofunctional alteration and inflammatory response (IM). IL-33 seems to be a compensatory protective factor on this inflammatory cascade. 5-LOX and PAFr show a likely secondary role on IM pathophysiology. |
publishDate |
2008 |
dc.date.none.fl_str_mv |
2008 2012-10-01T16:30:37Z 2012-10-01T16:30:37Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
LIMA JUNIOR, R. C. P. Estudo morfofuncional e dos mediadores inflamatórios envolvidos na patogênese da mucosite intestinal induzida por irinotecano (CPT-11) em camundongos : papel da caspase-1, interleucina-18 e óxido nitríco. 200 f. Tese (Doutorado em Farmacologia) - Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2008. http://www.repositorio.ufc.br/handle/riufc/3843 |
dc.identifier.dark.fl_str_mv |
ark:/83112/001300000z1c8 |
identifier_str_mv |
LIMA JUNIOR, R. C. P. Estudo morfofuncional e dos mediadores inflamatórios envolvidos na patogênese da mucosite intestinal induzida por irinotecano (CPT-11) em camundongos : papel da caspase-1, interleucina-18 e óxido nitríco. 200 f. Tese (Doutorado em Farmacologia) - Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2008. ark:/83112/001300000z1c8 |
url |
http://www.repositorio.ufc.br/handle/riufc/3843 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
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reponame:Repositório Institucional da Universidade Federal do Ceará (UFC) instname:Universidade Federal do Ceará (UFC) instacron:UFC |
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Universidade Federal do Ceará (UFC) |
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UFC |
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UFC |
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Repositório Institucional da Universidade Federal do Ceará (UFC) |
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Repositório Institucional da Universidade Federal do Ceará (UFC) |
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Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC) |
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bu@ufc.br || repositorio@ufc.br |
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