RESUMO
PURPOSE: To analyze the effects of ischemic preconditioning (IPC) in the expression of apoptosis-related genes in rat small intestine subjected to ischemia and reperfusion. METHODS: Thirty anesthetized rats underwent laparotomy and were drive into five groups: control (CG); ischemia (IG); ischemia and reperfusion (IRG); IPC and ischemia (IG+IPC); IPC and ischemia and reperfusion (I/RG+IPC). Intestinal ischemia was performed by clamping the superior mesenteric artery for 60 minutes, whereas reperfusion lasted for 120 minutes. IPC was carried out by one cycle of 5 minutes of ischemia followed by 10 minutes of reperfusion prior to the prolonged 60-minutes-ischemia and 120-minutes-reperfusion. Thereafter, the rats were euthanized and samples of small intestine were processed for histology and gene expression. RESULTS: Histology of myenteric plexus showed a higher presence of neurons presenting pyknotic nuclei and condensed chromatin in the IG and IRG. IG+IPC and I/RG+IPC groups exhibited neurons with preserved volume and nuclei, along with significant up-regulation of the anti-apoptotic protein Bcl2l1 and down-regulation of pro-apoptotic genes. Moreover, Bax/Bcl2 ratio was lower in the groups subjected to IPC, indicating a protective effect of IPC against apoptosis. CONCLUSION: Ischemic preconditioning protect rat small intestine against ischemia/reperfusion injury, reducing morphologic lesions and apoptosis.
Assuntos
Proteínas Reguladoras de Apoptose/análise , Apoptose/genética , Precondicionamento Isquêmico/métodos , Jejuno/irrigação sanguínea , Jejuno/patologia , Traumatismo por Reperfusão/prevenção & controle , Animais , Proteínas Reguladoras de Apoptose/genética , Constrição , Regulação para Baixo , Células Endoteliais/patologia , Expressão Gênica , Masculino , Artéria Mesentérica Superior , Isquemia Mesentérica/genética , Isquemia Mesentérica/patologia , Distribuição Aleatória , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Valores de Referência , Reprodutibilidade dos TestesRESUMO
Abstract Purpose: To analyze the effects of ischemic preconditioning (IPC) in the expression of apoptosis-related genes in rat small intestine subjected to ischemia and reperfusion. Methods: Thirty anesthetized rats underwent laparotomy and were drive into five groups: control (CG); ischemia (IG); ischemia and reperfusion (IRG); IPC and ischemia (IG+IPC); IPC and ischemia and reperfusion (I/RG+IPC). Intestinal ischemia was performed by clamping the superior mesenteric artery for 60 minutes, whereas reperfusion lasted for 120 minutes. IPC was carried out by one cycle of 5 minutes of ischemia followed by 10 minutes of reperfusion prior to the prolonged 60-minutes-ischemia and 120-minutes-reperfusion. Thereafter, the rats were euthanized and samples of small intestine were processed for histology and gene expression. Results: Histology of myenteric plexus showed a higher presence of neurons presenting pyknotic nuclei and condensed chromatin in the IG and IRG. IG+IPC and I/RG+IPC groups exhibited neurons with preserved volume and nuclei, along with significant up-regulation of the anti-apoptotic protein Bcl2l1 and down-regulation of pro-apoptotic genes. Moreover, Bax/Bcl2 ratio was lower in the groups subjected to IPC, indicating a protective effect of IPC against apoptosis. Conclusion: Ischemic preconditioning protect rat small intestine against ischemia/reperfusion injury, reducing morphologic lesions and apoptosis.
Assuntos
Animais , Masculino , Traumatismo por Reperfusão/prevenção & controle , Apoptose/genética , Precondicionamento Isquêmico/métodos , Proteínas Reguladoras de Apoptose/análise , Jejuno/irrigação sanguínea , Jejuno/patologia , Valores de Referência , Distribuição Aleatória , Regulação para Baixo , Expressão Gênica , Reprodutibilidade dos Testes , Ratos Wistar , Artéria Mesentérica Superior , Constrição , Células Endoteliais/patologia , Proteínas Reguladoras de Apoptose/genética , Reação em Cadeia da Polimerase em Tempo Real , Isquemia Mesentérica/genética , Isquemia Mesentérica/patologiaRESUMO
Abstract Purpose: To investigate the gene expression related to inflammation on mice subjected to intestinal ischemia and reperfusion (I/R) and treated with ischemic preconditioning (IPC). Methods: Thirty rats (EPM-Wistar), distributed in five groups of six animals each, were underwent anesthesia and laparotomy. The ischemia time was standardized in 60 minutes and the reperfusion time 120 minutes. IPC was standardized in 5 minutes of ischemia followed by 10 minutes of reperfusion accomplished before I/R. The control group was submitted only to anesthesia and laparotomy. The other groups were submitted to ischemia, I/R, ischemia + IPC and I/R + IPC. It was collected a small intestine sample to analyses by Quantitative Polymerase Chain Reaction in real Time (RT-qPCR) and histological analyses. It was studied 27 genes. Results: The groups that received IPC presented downregulation of genes, observed in of genes in IPC+ischemia group and IPC+I/R group. Data analysis by clusters showed upregulation in I/R group, however in IPC groups occurred downregulation of genes related to inflammation. Conclusion: The ischemia/reperfusion promoted upregulation of genes related to inflammation, while ischemic preconditioning promoted downregulation of these genes.
Assuntos
Animais , Masculino , Traumatismo por Reperfusão/prevenção & controle , Expressão Gênica/fisiologia , Precondicionamento Isquêmico/métodos , Inflamação/prevenção & controle , Intestino Delgado/irrigação sanguínea , Valores de Referência , Fatores de Tempo , Traumatismo por Reperfusão/genética , Regulação para Baixo/fisiologia , Regulação para Cima/fisiologia , Reprodutibilidade dos Testes , Resultado do Tratamento , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Isquemia Mesentérica/genética , Isquemia Mesentérica/prevenção & controle , Inflamação/genéticaRESUMO
PURPOSE: To investigate the gene expression related to inflammation on mice subjected to intestinal ischemia and reperfusion (I/R) and treated with ischemic preconditioning (IPC). METHODS: Thirty rats (EPM-Wistar), distributed in five groups of six animals each, were underwent anesthesia and laparotomy. The ischemia time was standardized in 60 minutes and the reperfusion time 120 minutes. IPC was standardized in 5 minutes of ischemia followed by 10 minutes of reperfusion accomplished before I/R. The control group was submitted only to anesthesia and laparotomy. The other groups were submitted to ischemia, I/R, ischemia + IPC and I/R + IPC. It was collected a small intestine sample to analyses by Quantitative Polymerase Chain Reaction in real Time (RT-qPCR) and histological analyses. It was studied 27 genes. RESULTS: The groups that received IPC presented downregulation of genes, observed in of genes in IPC+ischemia group and IPC+I/R group. Data analysis by clusters showed upregulation in I/R group, however in IPC groups occurred downregulation of genes related to inflammation. CONCLUSION: The ischemia/reperfusion promoted upregulation of genes related to inflammation, while ischemic preconditioning promoted downregulation of these genes.
Assuntos
Expressão Gênica/fisiologia , Inflamação/prevenção & controle , Intestino Delgado/irrigação sanguínea , Precondicionamento Isquêmico/métodos , Traumatismo por Reperfusão/prevenção & controle , Animais , Regulação para Baixo/fisiologia , Inflamação/genética , Masculino , Isquemia Mesentérica/genética , Isquemia Mesentérica/prevenção & controle , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Valores de Referência , Traumatismo por Reperfusão/genética , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do Tratamento , Regulação para Cima/fisiologiaRESUMO
PURPOSE: To study the expression of HER2, p53 and Ki67 proteins in cystoplasties. METHODS: Sixty rats were distributed randomly into three groups of 20 animals. Bladder augmentation was held to increase with ileum (Group I), colon (Group II) and stomach (Group III). Tissue samples of neobladder was collected from each rat to its own control. The animals were sacrificed after 12 weeks. The neobladder was withdrawn for immunohistochemistry analysis of p53, HER2 and Ki67 expression. Wilcoxon and Mann-Whitney tests were used for statistical study. RESULTS: There were no significant changes in the expression of p53 and HER2 proteins. It was observed significant increase (p<0.0001) in Ki67 expression in all groups, when compared with their respective controls. When the study groups were compared with each other, there was increase of cell proliferation in the largest gastrocystoplasties in respect of ileocystoplasties (p=0.004) and colocystoplasties (p=0.003). CONCLUSION: We observed significant increase of cell proliferation characterized by Ki67 protein in the digestive tract of the ileocystoplasties, the colocystoplasties and the gastrocystoplasties and this increase was significantly greater in gastrocystoplasties.
Assuntos
Colo/metabolismo , Mucosa Gástrica/metabolismo , Íleo/metabolismo , Antígeno Ki-67/metabolismo , Trato Gastrointestinal Inferior/cirurgia , Receptor ErbB-2/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Bexiga Urinária/cirurgia , Animais , Colo/transplante , Íleo/transplante , Imuno-Histoquímica , Trato Gastrointestinal Inferior/metabolismo , Ratos Wistar , Estatísticas não Paramétricas , Estômago/transplante , Bexiga Urinária/metabolismoRESUMO
PURPOSE: To study the expression of HER2, p53 and Ki67 proteins in cystoplasties. METHODS: Sixty rats were distributed randomly into three groups of 20 animals. Bladder augmentation was held to increase with ileum (Group I), colon (Group II) and stomach (Group III). Tissue samples of neobladder was collected from each rat to its own control. The animals were sacrificed after 12 weeks. The neobladder was withdrawn for immunohistochemitry analysis of p53, HER2 and Ki67 expression. Wilcoxon and Mann-Whitney tests were used for statistical study. RESULTS: There were no significant changes in the expression of p53 and HER2 proteins. It was observed significant increase (p<0.0001) in Ki67 expression in all groups, when compared with their respective controls. When the study groups were compared with each other, there was increase of cell proliferation in the largest gastrocystoplasties in respect of ileocystoplasties (p=0.004) and colocystoplasties (p=0.003). CONCLUSION: We observed significant increase of cell proliferation characterized by Ki67 protein in the digestive tract of the ileocystoplasties, the colocystoplasties and the gastrocystoplasties and this increase was significantly greater in gastrocystoplasties.
