ABSTRACT
Abstract Purpose: To evaluate the effect of hyperbaric oxygenation (HBO) on the expression of the genes antioxidant glutathione peroxidase 4 (Gpx4) and lactoperoxidase (Lpo) in the lung of mice subjected to intestinal ischemia and reperfusion (IIR). Methods: Control group (CG) in which were subjected to anesthesia, laparotomy and observation for 120 minutes; an ischemia and reperfusion group (IRG) subjected to anesthesia, laparotomy, small bowel ischemia for 60 minutes and reperfusion for 60 minutes; and three groups treated with HBO during ischemia (HBOG + I), during reperfusion (HBOG + R) and during ischemia and reperfusion (HBOG + IR). Studied 84 genes of oxidative stress by the method (RT-qPCR). 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: Gpx4 and Lpo were hiperexpressed on IRG, showing a correlation with these genes with lung oxidative stress. Treated with HBO, there was a significant reduction on genic expression on HBOG+I. Conclusion: Hyperbaric oxygenation showed to be associated with decreased expression of these antioxidant genes, suggesting a beneficial effect on the mechanism of pulmonary oxidative stress whenever applied during the ischemia.
Subject(s)
Animals , Rats , Reperfusion Injury/metabolism , Oxidative Stress/genetics , Glutathione Peroxidase/metabolism , Hyperbaric Oxygenation/methods , Lactoperoxidase/genetics , Lung/metabolism , Oxidative Stress/drug effects , Disease Models, Animal , Intestines/blood supply , Ischemia/metabolism , Antioxidants/metabolism , Antioxidants/pharmacologyABSTRACT
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.
Subject(s)
Animals , Male , Mice , Reperfusion Injury/prevention & control , Gene Expression , Oxidative Stress/genetics , Hyperbaric Oxygenation/methods , Intestines/blood supply , Reperfusion Injury/metabolism , Polymerase Chain Reaction , Oxidative Stress/drug effects , NADPH Oxidases/metabolism , Coronary Vessels/enzymology , Disease Models, Animal , Heart , Heart Diseases , Ischemia/metabolism , Mice, Inbred C57BL , Antioxidants/metabolism , Antioxidants/pharmacologyABSTRACT
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 2.
Subject(s)
Animals , Mice , Ischemia/pathology , Oxidative Stress , Lung/anatomy & histology , Mice/classificationABSTRACT
FUNDAMENTO: O fenômeno da isquemia e reperfusão intestinal é um evento frequente na clínica e está associado a repercussões deletérias em órgãos a distância, em especial ao coração. OBJETIVO: Investigar a expressão gênica do estresse oxidativo e defesa antioxidante no coração de camundongos isogênicos, submetidos a isquemia e reperfusão intestinal (IR). MÉTODOS: Doze camundongos (C57BL/6) foram distribuídos em dois grupos: Grupo IR (GIR) com 60 min de oclusão da artéria mesentérica superior, seguidos de 60 min de reperfusão. Grupo Controle (GC) submetidos a anestesia e a laparotomia sem o procedimento de IR observados por 120 min. As amostras de intestino e coração foram processadas pelo método (RT-qPCR / Reverse transcriptase - quantitative Polymerase Chain Reaction) para determinar a expressão gênica de 84 genes relacionados ao estresse oxidativo ("t" de Student, p < 0,05). RESULTADOS: Observou-se no tecido intestinal (GIR) uma expressão significantemente aumentada em 65 (74,71%) genes em relação ao tecido normal (GC), e 37 (44,04%) genes estiveram hiperexpressos (maior que três vezes o limiar permitido pelo algoritmo). No tocante aos efeitos da I/R intestinal a distância no tecido cardíaco verificou-se a expressão significantemente aumentada de 28 genes (33,33%), mas somente oito genes (9,52%) se hiperexpressaram três vezes acima do limiar. Quatro (7,14%) desses oito genes se expressaram simultaneamente nos tecidos intestinal e cardíaco. No GIR notaram-se cardiomiócitos com núcleos de menor tamanho, picnóticos, ricos em heterocromatina e raros nucléolos, indicando sofrimento cardíaco. CONCLUSÃO: A I/R intestinal promoveu a hiperexpressão estatisticamente significante de oito genes associados ao ...
BACKGROUND: Intestinal ischemia-reperfusion is a frequent clinical event associated to injury in distant organs, especially the heart. OBJECTIVE: To investigate the gene expression of oxidative stress and antioxidant defense in the heart of inbred mice subjected to intestinal ischemia and reperfusion (IR). METHODS: Twelve mice (C57BL / 6) were assigned to: IR Group (GIR) with 60 minutes of superior mesenteric artery occlusion followed by 60 minutes of reperfusion; Control Group (CG) which underwent anesthesia and laparotomy without IR procedure and was observed for 120 minutes. Intestine and heart samples were processed using the RT-qPCR / Reverse transcriptase-quantitative Polymerase Chain Reaction method for the gene expression of 84 genes related to oxidative stress and oxidative defense (Student's "t" test, p < 0.05). RESULTS: The intestinal tissue (GIR) was noted to have an up-regulation of 65 genes (74.71%) in comparison to normal tissue (CG), and 37 genes (44.04%) were hyper-expressed (greater than three times the threshold allowed by the algorithm). Regarding the remote effects of intestinal I/R in cardiac tissue an up-regulation of 28 genes (33.33%) was seen, but only eight genes (9.52%) were hyper-expressed three times above threshold. Four (7.14%) of these eight genes were expressed in both intestinal and cardiac tissues. Cardiomyocytes with smaller and pyknotic nuclei, rich in heterochromatin with rare nucleoli, indicating cardiac distress, were observed in the GIR. CONCLUSION: Intestinal I/R caused a statistically significant over expression of 8 genes associated with oxidative stress in remote myocardial tissue. .
Subject(s)
Animals , Male , Gene Expression/genetics , Intestine, Small/blood supply , Myocardium/metabolism , Oxidative Stress/genetics , Reperfusion Injury/metabolism , Antioxidants/metabolism , Disease Models, Animal , Intestine, Small/metabolism , Ischemia/genetics , Ischemia/metabolism , Random Allocation , Reverse Transcriptase Polymerase Chain Reaction , Reperfusion Injury/genetics , Time Factors , Up-Regulation/geneticsABSTRACT
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.
Subject(s)
Animals , Male , Mice , Gene Expression/genetics , Intestine, Small , Kidney , Oxidative Stress/genetics , Reperfusion Injury/genetics , Antioxidants/metabolism , Down-Regulation/genetics , Intestine, Small/metabolism , Intestine, Small/physiopathology , Kidney/metabolism , Kidney/physiopathology , Random Allocation , Reverse Transcriptase Polymerase Chain Reaction , Time Factors , Up-Regulation/geneticsABSTRACT
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.
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.
Subject(s)
Animals , Male , Mice , Intestine, Small/blood supply , Intestine, Small/surgery , Ischemia/genetics , Oxidative Stress/genetics , Reperfusion Injury/metabolism , Antioxidants/metabolism , Down-Regulation/genetics , Gene Expression Profiling , Ischemia/metabolism , Multigene Family/genetics , Random Allocation , Real-Time Polymerase Chain Reaction , Reperfusion Injury/genetics , Time Factors , Up-Regulation/geneticsABSTRACT
JUSTIFICATIVA E OBJETIVOS: O objetivo deste estudo foi destacar a importância da associação do exame físico acurado à exames complementares, na identificação de doenças raras como a amiloidose cardíaca. RELATO DO CASO: Paciente do sexo masculino, 58 anos, com história de dispneia, cansaço e edema nos membros inferiores (MMII) há aproximadamente nove meses, com evolução e piora rápidas do quadro clínico, passando do grau lll para o grau lV segundo a NYHA (Associação de Cardiologiade Nova York), sendo realizados cateterismo e ecocardiografia (ECO) para avaliar a função hemodinâmica. Ao realizarem-se os exames, foram observados hipertrofia de ventrículo esquerdo (VE) com moderada disfunção sistólica e diastólica; discreta insuficiência mitral e dominância coronariana direita. O paciente foi submetido ao tratamento convencional para o quadro de insuficiência cardíaca congestiva (ICC) moderada, como o uso de digital, diurético de alça associado à um poupador de potássio, beta-bloqueador e inibidor da enzima de conversão da angiotensina (IECA). Seis meses depois, apresentou piora do quadro clínico com evolução para ICC de grau IV. Realizou-se novo ECO, que apresentou hipertrofia do VE, hipocinesia difusa e miocárdio com aspecto infiltrativo e refringente, quadro característico e compatível com os achados sugestivos de amiloidose cardíaca; doença confirmada pela biopsia do miocárdio. CONCLUSÃO: A relevância da avaliação clínica deste caso, em que se destaca a rápida evolução do quadro geral para piora acentuada, após o uso de medicamentos sabidamente úteis no tratamento da ICC, porém, que podem contribuir para piora nos casos de amiloidose, como, por exemplo, diuréticos digitálicos e IECA. Em associação aos exames complementares, com destaque para o ecocardiograma, foi de extrema importância para a aproximação do diagnóstico definitivo, obtido por meio do estudo anatomopatológico.
BACKGROUND AND OBJECTIVES: The objective of this study was to highlight the importance of the combination of physical examination and complementary tests for accurate identification of rare diseases such as cardiac amyloidosis. CASE REPORT: Male patient, 58 years, with a history of dyspnea, fatigue and swelling in the lower limbs (LL) with development and worsening fast, being performed catheterization and echocardiography (ECHO), to assess hemodynamic functions. When carrying out the examination, we observed hypertrophy of the left ventricle (LV) with moderate systolic and diastolic dysfunction, mild mitral regurgitation and right coronary dominance. The patient was undergoing treatment for the conventional framework of moderate congestive heart failure (CHF). Six months later, presented with worsening of clinical development for CCI grade IV. There was new ECO, which showed the LV hypertrophy, and diffuse hypokinesis with myocardial infiltration and refractile appearance, characteristic of this framework cardiac amyloidosis; disease confirmed by myocardial biopsy. CONCLUSION: The accuracy of physical examination, coupled with the complementary examinations, was of extreme importance for the approximation of the final diagnosis obtained by means of histopathological examination.