Perfil da expressão gênica do estresse oxidativo no pulmão de camundongos isogênicos após lesão de isquemia e reperfusão em intestino delgado
Arquivos
Data
2014
Autores
Ikejiri, Adauto Tsutomu 
Orientadores
Fagundes, Djalma José
Tipo
Dissertação de mestrado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
Objetivo: determinar o perfil de expressão gênica associada ao estresse oxidativo e a defesa antioxidante no tecido pulmonar de camundongos submetidos à isquemia e reperfusão intestinal. Métodos: doze camundongos isogênicos do sexo masculino (C57BL/6) foram distribuídos aleatoriamente em dois grupos: Grupo Controle (GC) com animais submetidos à anestesia, laparotomia e observação de 120 minutos; Grupo isquemia/reperfusão (GIR), submetido à anestesia, laparotomia, isquemia por 60 minutos de intestino delgado e 60 minutos de reperfusão. Um conjunto de amostra dos seis animais de cada grupo foi submetido a transcrição reversa reação em cadeia da polimerase quantitativa (RTqPCR) em tempo real, para determinação da expressão gênica do estresse oxidativo e de defesa antioxidante. Todos os genes que foram hiperexpressos ou hipoexpressos acima de três vezes o limiar permitido pelo algoritmo [2^(ΔΔCt)] foram considerados biologicamente relevantes. Resultados: nossos dados mostram que dos 84 genes no pulmão relacionados ao estresse oxidativo 67 (79,7%) deles se expressaram positivamente e 17 (20,2%) se expressaram negativamente. Apenas dois genes (2,3%): Lpo Lactoperoxidase (+3,51) e Gpx4 Glutationa peroxidase (+4,10), foram expressos além do triplo do limiar do ciclo, permitido pelo algoritmo utilizado, enquanto nenhum dos genes que se expressaram negativamente ultrapassou o triplo do limiar do ciclo. Conclusão: a lesão de isquemia/reperfusão intestinal promove um perfil de expressão positiva de genes relacionados ao estresse oxidativo sobre órgãos à distância, sugerindo que ativam as vias de sinalização implicadas na sobrevivência e manutenção da integridade do genoma de células do tecido pulmonar.
Purpose: To determine the gene expression profile associated with oxidative stress and antioxidant defense in the lung tissue of mice subjected to intestinal ischemia and reperfusion. Methods: Twelve male, inbred mice (C57BL/6) were randomly assigned to one of two groups. The control group (GC) underwent anesthesia and laparotomy and was observed for 120 minutes; the ischemia/reperfusion group (GIR) was subjected to anesthesia, laparotomy, and ischemia of the small intestine for 60 minutes and to 60 minutes of reperfusion. A pool of six mice from each group was subjected to a real time reverse transcription quantitative polymerase chain reaction (RTqPCR), to analyze the oxidative stress and antioxidant defense genes. All genes that were upregulated or downregulated greater than threefold, based on the algorithm [2^(ΔΔCt)], were considered to be biologically meaningful. Results: Out of a total of 84 genes in the lung that are related to oxidative stress, 67 (79.7%) were upregulated and 17 (20.2%) were downregulated. Only two genes (2.3%), Lpo (Lactoperoxidase) (+3.51) and Gpx4 (Glutathione peroxidase) (+4.10), were expressed above the threefold threshold, while none of the downregulated genes were expressed outside of this threshold. Conclusion: The intestinal ischemia/reperfusion injury promoted a gene expression profile consisting of the positive expression of oxidative genes in a remote organ. This suggests that activate signaling pathways are implicated in both cell survival and the maintenance of genome integrity in the lung.
Purpose: To determine the gene expression profile associated with oxidative stress and antioxidant defense in the lung tissue of mice subjected to intestinal ischemia and reperfusion. Methods: Twelve male, inbred mice (C57BL/6) were randomly assigned to one of two groups. The control group (GC) underwent anesthesia and laparotomy and was observed for 120 minutes; the ischemia/reperfusion group (GIR) was subjected to anesthesia, laparotomy, and ischemia of the small intestine for 60 minutes and to 60 minutes of reperfusion. A pool of six mice from each group was subjected to a real time reverse transcription quantitative polymerase chain reaction (RTqPCR), to analyze the oxidative stress and antioxidant defense genes. All genes that were upregulated or downregulated greater than threefold, based on the algorithm [2^(ΔΔCt)], were considered to be biologically meaningful. Results: Out of a total of 84 genes in the lung that are related to oxidative stress, 67 (79.7%) were upregulated and 17 (20.2%) were downregulated. Only two genes (2.3%), Lpo (Lactoperoxidase) (+3.51) and Gpx4 (Glutathione peroxidase) (+4.10), were expressed above the threefold threshold, while none of the downregulated genes were expressed outside of this threshold. Conclusion: The intestinal ischemia/reperfusion injury promoted a gene expression profile consisting of the positive expression of oxidative genes in a remote organ. This suggests that activate signaling pathways are implicated in both cell survival and the maintenance of genome integrity in the lung.
Descrição
Citação
IKEJIRI, Adauto Tsutomu. Perfil da expressão gênica do estresse oxidativo no pulmão de camundongos isogênicos após lesão de isquemia e reperfusão em intestino delgado. 2014. 108 f. Dissertação (Mestrado em Ciências) – Escola Paulista de Medicina, Universidade Federal de São Paulo. São Paulo, 2014.