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.
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.
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.
PURPOSE: To introduce a model of equipment for inhalation anesthesia in rats that offers better control of both flow and losses of ether during induction, maintenance, and recuperation. METHODS: The equipment consists of an air compressor with two outlets, a closed glass induction chamber, a glass reservoir for the anesthetic agent, a pediatric inhalation mask, a three-way stopcock, a Y-connector, and urinary catheters. Three hundred Wistar rats (Rattus norvegicus albinus) were given inhalation anesthesia. The evaluated parameters were equipment operation, duration of each phase of anesthesia, corneal reflex, muscular tonus, respiration during induction and maintenance, and volume of anesthesia. RESULTS: The average time taken for induction was 7.3 minutes; the average anesthetic recuperation time was 6.4 minutes. The amount of anesthetic used varied according to the weight of the animal, with the average volume of ether used being 6.5ml/hour. The availability of oxygen (room air) decreased the recuperation time and averted both respiratory depression and insufficient depth of anesthesia. CONCLUSION: The proposed equipment is practical, inexpensive, and allows for satisfactory control of anesthetic parameters during the entire procedure, making inhalation anesthesia in rats safe and essentially complication free.
OBJETIVO: Apresentar modelo de equipamento para anestesia inalatória em ratos, com melhor controle do fluxo, das perdas de anestésico na indução, da manutenção e da recuperação anestésica. MÉTODOS: O equipamento é constituído por compressor de ar com saída dupla, câmara de vidro fechada para indução, reservatório de vidro para o agente anestésico, máscara de inalação pediátrica, torneira de três vias, conexão em "Y" e sondas uretrais. Trezentos ratos Wistar (Rattus norvegicus albinus) foram anestesiados por via inalatória. Os parâmetros avaliados foram: viabilidade do equipamento, tempo de cada fase, reflexo corneano, tônus muscular e movimentos respiratórios na indução e na manutenção anestésica, além do volume de anestésico. RESULTADOS: A média do tempo gasto na indução anestésica foi de 7,3 minutos. A média do tempo de recuperação anestésica foi de 6,4 minutos. A média do volume de éter utilizado foi de 6,5 ml/h. Não foram observadas neste experimento morte dos animais, aumento excessivo de secreção traqueobrônquica ou interrupção da operação por superficialização anestésica. CONCLUSÃO: O equipamento proposto é prático, possui baixo custo e permite o controle adequado dos parâmetros de controle da anestesia durante todo o procedimento, tornando a anestesia inalatória em ratos segura e praticamente isenta de complicações.
PURPOSE: To study morphologic and histochemical alterations arising at the ileocystoplasty site. METHODS: Sixteen Wistar female rats were subjected to ileocystoplasty and sacrificed after eight weeks. Material collected was divided into four groups for histological and histochemical studies: Group I (control) - isolated ileum segment removed during ileocystoplasty; Group II - ileoileal anastomosis; Group III - ileovesical anastomosis and Group IV - ileal segment from the neobladder. Histological and histochemical study assessed dysplasia, metaplasia, acute and chronic inflammation, fibrosis, atrophy, hypertrophy, total mucins, sialomucins and sulfomucins. The non-parametric Wilcoxon and Mann-Whitney tests were employed in statistical analysis. RESULTS: None of the groups presented dysplasia. Acute inflammation and atrophy occurred in Groups II, III and IV, not reaching statistical significance. Metaplasia was significant only in Group III (p=0.012). Chronic inflammation, fibrosis and hypertrophy were significant in Groups II, III and IV. There was a significant increase in total mucin content in Group IV (p=0.014) and a reduction in Group III (p=0.016). Increases in sialomucins were observed in samples for Groups III (p=0.003) and IV (p=0.002) along with reduced sulfomucins in samples from Groups III (p=0.013) and IV (p=0.008). CONCLUSION: Ileocystoplasty in female rats caused squamous metaplasia, chronic inflammatory infiltration, fibrosis, hypertrophy, increase in sialomucin content, reduction in sulfomucins, and alterations in total mucin content with statistical significance, as well acute inflammatory infiltration and muscular atrophy with less intensity.
OBJETIVO: Estudar alterações morfológicas e histoquímicas nas ileocistoplastias em ratos fêmea. MÉTODOS: 16 ratos fêmea foram submetidos à ileocistoplastia, sacrificadas após oito semanas. O material coletado foi e dividido em quatro grupos para análise morfológica e histoquímica: Grupo I (controle) biópsia intestinal no momento da cirurgia; Grupo II - anastomose íleo-ileal; Grupo III - anastomose íleo-vesical e Grupo IV - segmento intestinal da neobexiga. Os parâmetros avaliados foram: displasia, metaplasia, processo inflamatório agudo e crônico, fibrose, atrofia, hipertrofia, conteúdo total de mucinas, sialomucinas e sulfomucinas. Utilizou-se os testes não-paramétricos de Wilcoxon e Mann-Whitney para estudo estatístico. RESULTADOS: Não houve displasia. Processo inflamatório agudo e atrofia ocorreram nos grupos II, III e IV, sem significância estatística. Metaplasia com significância estatística ocorreu somente no grupo III (p=0.012). Processo inflamatório crônico, fibrose e hipertrofia foram significantes nos grupos II, III e IV. Observou-se aumento significante no conteúdo total de mucinas no grupo IV (p=0.014) e redução no grupo III (p=0.013). Aumento significante de sialomucinas foi observado nos grupos III (p=0.003) e IV (p=0.002) e redução significante das sulfomucinas nos grupos III (p=0.013) e IV (p=0.008). CONCLUSÃO: Nas ileocistoplastias em ratos fêmea observou-se metaplasia escamosa, processo inflamatório crônico, fibrose, hipertrofia, aumento do conteúdo de sialomucinas, redução das sulfomucinas e alterações no conteúdo total de mucinas com significância estatística, bem como atrofia e processo inflamatório agudo em menor intensidade.