Expression of oxidative stress and antioxidant defense genes in the kidney of inbred mice after intestinal ischemia and reperfusion

PURPOSE: To determine the gene expressions profile related to the oxidative stress and the antioxidant response in the kidneys of mice subjected to intestinal ischemia and reperfusion. METHODS: Twelve inbred mice (C57BL/6) were randomly assigned to one of two groups: the control group (CG) underwent anesthesia and was observed for 120 min and the ischemia/reperfusion group (IRG), animals were anesthetized and subjected to laparotomy and ischemia for 60 minutes followed by 60 minutes of reperfusion. The expressions of 84 genes from the kidney were determined by the Reverse Transcription qualitative Polymerase Chain Reaction (RT-qPCR). All genes that were up regulated by more than threefold using the algorithm [2(ΔΔCt)] were considered statically significant (p<0.05). RESULTS: In the IRG group 29 (34.52%) of 84 genes, were up regulated by more than threefold. The genes that were differentially up regulated in the glutathione peroxidase cluster (10 genes): were Gpx2 and Gpx7. The genes that were up regulated in the peroxidase cluster (16 genes) were following: Duox1, Epx, Lpo, Mpo, Ptgs2, Rag2, Serpinb1b, Tmod1 and Tpo. The genes that up regulated in the reactive oxygen species cluster (16 genes): Il19, Il22, Nos2, Nox1, Noxa1, Noxo1, Recql4 and Sod2. The genes that were up regulated in the oxidative stress cluster (22 genes) were: Mpp4, Nudt15, Upc3 and Xpa. The genes that were up regulated in the oxygen carriers cluster (12 genes) were: Hbq1, Mb, Ngb, Slc38a1 and Xirp1. The peroxiredoxins genes (10) showed no consistent differential regulation. CONCLUSION: The genes related to oxidative stress and antioxidant defense showed increased expression in renal tissue trigged intestinal ischemia and reperfusion.São Paulo Federal University Postgraduate Program in Interdisciplinary Surgical SciencesMato Grosso do Sul Federal University Surgery DepartmentGrande Dourados Federal University Medical SchoolUFGD Medical SchoolUNIFESP Surgery Department Surgical Techniques and Experimental Surgery DivisionSão Paulo Federal University Postgraduate Program in Interdisciplinary Surgical SciencesUNIFESP, Surgery Department Surgical Techniques and Experimental Surgery DivisionSciEL

Gene expression profile of oxidative stress in the lung of inbred mice after intestinal ischemia/reperfusion injury

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 (CG) underwent anesthesia and laparotomy and was observed for 120 minutes; the ischemia/reperfusion group (IRG) 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 reverse transcription-quantitative polymerase chain reaction (RT-qPCR) to analyze the oxidative stress and antioxidant defense genes. All genes that were up-regulated or down-regulated greater than three-fold, based on the algorithm [2Federal University of Grande Dourados Medical SchoolUFGD Medical SchoolFederal University of Mato Grosso do Sul Surgery DepartmentUniversitary Center Sao CamiloUNIFESP Surgery DepartmentUNIFESP, Surgery DepartmentSciEL

Hyperbaric oxygenation and the genic expression related to oxidative stress in the heart of mice during intestinal ischemia and reperfusion

<div><p>Abstract Purpose: To investigate the effects of hyperbaric oxygenation (HBO) on intestinal ischemia and reperfusion (IR) injury, we evaluated the expression of 84 genes related to oxidative stress and the antioxidant response in mouse hearts. Methods: Four groups were subjected to 60 minutes of intestinal ischemia followed by 60 minutes of reperfusion: IRG, ischemia and reperfusion group without HBO; HBO-IG, which received HBO during ischemia; HBO-RG, which received HBO during reperfusion; and HBO-IRG, which received HBO during ischemia and reperfusion. The control group (CG) underwent anesthesia and laparotomy and was observed for 120 minutes. The (RT-qPCR) method was applied. Genes with expression levels three times below or above the threshold cycle were considered significantly hypoexpressed or hyperexpressed, respectively (Student’s t-test p<0.05). Results: Eight genes (9.52%) were hyperexpressed in the IRG. When the HBO groups were compared to the IRG, we found a decrease in the expression of eight genes in the HBO-IG, five genes in the HBO-RG, and seven genes in the HBO-IRG. Conclusion: The reduction in the expression of genes related to oxidative stress and antioxidant defense following HBO in mouse hearts resulting from intestinal IR injury was more favorable during the ischemic period than during the reperfusion period.</p></div

Oxidative stress gene expression profile in inbred mouse after ischemia/reperfusion small bowel injury Perfil da expressão gênica do estresse oxidativo em camundongos isogênicos após lesão de isquemia e reperfusão intestinal

PURPOSE: To determine the profile of gene expressions associated with oxidative stress and thereby contribute to establish parameters about the role of enzyme clusters related to the ischemia/reperfusion intestinal injury. METHODS: Twelve male inbred mice (C57BL/6) were randomly assigned: Control Group (CG) submitted to anesthesia, laparotomy and observed by 120min; Ischemia/reperfusion Group (IRG) submitted to anesthesia, laparotomy, 60min of small bowel ischemia and 60min of reperfusion. A pool of six samples was submitted to the qPCR-RT protocol (six clusters) for mouse oxidative stress and antioxidant defense pathways. RESULTS: On the 84 genes investigated, 64 (76.2%) had statistic significant expression and 20 (23.8%) showed no statistical difference to the control group. From these 64 significantly expressed genes, 60 (93.7%) were up-regulated and 04 (6.3%) were down-regulated. From the group with no statistical significantly expression, 12 genes were up-regulated and 8 genes were down-regulated. Surprisingly, 37 (44.04%) showed a higher than threefold up-regulation and then arbitrarily the values was considered as a very significant. Thus, 37 genes (44.04%) were expressed very significantly up-regulated. The remained 47 (55.9%) genes were up-regulated less than three folds (35 genes - 41.6%) or down-regulated less than three folds (12 genes - 14.3%). CONCLUSION: The intestinal ischemia and reperfusion promote a global hyper-expression profile of six different clusters genes related to antioxidant defense and oxidative stress.<br>OBJETIVO: Determinar o perfil de expressão dos genes associados com estresse oxidativo e contribuir para estabelecer parâmetros sobre o papel das familias de enzimas relacionadas com a lesão de isquemia / reperfusão intestinal. MÉTODOS: Doze camundongos machos isogênicos (C57BL/6) foram distribuídos aleatoriamente: Grupo Controle (CG) submetido à laparotomia anestesia, e observado por 120min; Grupo isquemia/reperfusão (IRG) submetido à anestesia, laparotomia, 60min de isquemia do intestino delgado e 60min de reperfusão. Um pool dos seis camundongos de cada grupo foi submetido ao protocolo de qPCR-RT (seis famílias) para o estresse oxidativo e defesa antioxidante. RESULTADOS: Dos 84 genes investigados, 64 (76,2%) tiveram expressão estatística significante e 20 (23,8%) não apresentaram diferença estatística com o grupo controle. Dos 64 genes expressos de forma significante, 60 (93,7%) foram hiper-expressos e 04 (6,3%) foram hipo-expressos. Do grupo sem expressão estatisticamente significante, 12 genes foram hiper e 8 genes foram hipo-expressos. Surpreendentemente, 37 (44,04%) apresentaram expressão três maior que o limiar de normalidade e arbitrariamente os valores foram considerados como altamente significantes. Assim, 37 genes (44,04%) foram hiper-expressos de modo muito significante. Nos demais, 47 (55,9%) dos genes foram hiper-expressos menos de três vezes (35 genes - 41,6%) ou hipo-expressos menos de três vezes(12 genes - 14,3%). CONCLUSÃO: A isquemia e reperfusão intestinal promoveu um perfil de hiper-expressão global das seis familias de genes relacionados com estresse oxidativo antioxidante e defesa antioxidante