Análise da expressão de biomarcadores inflamatórios em enxertos utilizados em pacientes diabéticos e não diabéticos submetidos à revascularização do miocárdio
Análise da expressão de biomarcadores inflamatórios em enxertos utilizados em pacientes diabéticos e não diabéticos submetidos à revascularização do miocárdio
Data
2016
Autores
Silva, Ana Catarina Romano e
Journal Title
Journal ISSN
Volume Title
Publisher
Instituto Nacional de Cardiologia
Resumo
Introdução: Sabe-se que os pacientes diabéticos apresentam uma progressão da doença aterosclerótica mais acentuada que não-diabeticos. A inflamação, o estresse oxidativo e a deposição de matriz extracelular na parede do vaso são descritos como pontos-chave na fisiopatologia da aterosclerose, em pesquisas feitas principalmente com modelos animais. Existem poucos estudos analisando estes parâmetros em artérias de médio calibre de pacientes diabéticos e comparando com não-diabeticos. Objetivos: Avaliar a presença dos marcadores inflamatórios fator de necrose tumoral-α (TNF-α), fator de crescimento transformador-β1 (TGF-β1), fator nuclear kappa B (NF-κB), as enzimas superóxido dismutase (SOD), catalase (CAT) e óxido nítrico sintase na forma induzida (iNOS) e do colágeno e fibras elásticas presentes na parede vascular, do enxerto utilizado na CRVM em DM e NDM buscando relações entre resultados obtidos com parâmetros clínicos dos grupos estudados. Metodologia: foram incluídas no estudo amostras de enxerto arterial de 23 pacientes masculinos que realizaram CRVM no Instituto Nacional de Cardiologia entre dezembro de 2014 e dezembro de 2015. As análises de expressão dos marcadores inflamatórios foram realizadas através de quantificação relativa por reação em cadeia da polimerase (qPCR) em tempo real e imunohistoquímica (IHQ). Foram realizadas ainda colorações para colágeno e fibras elásticas no tecido de enxerto. Resultados: Os dados clínicos demonstraram que os grupos de DM e NDM eram clinicamente semelhantes, com exceção da existência de diabetes mellitus. A mediana da idade do grupo DM foi de 59 anos e do grupo NDM foi de 64,5 anos. Os pacientes diabéticos apresentaram maior elevação de troponina T no pós-operatório com mediana de 5,81 mg/dL enquanto o grupo NDM apresentou mediana de 2,62 mg/dL. Os resultados obtidos por qPCR não mostraram diferença estatística entre os grupos analisados, embora tenha havido um aumento na expressão de CAT de 84% no grupo de DM quando comparado com os NDM. Os marcadores NF-kB, SOD e TGF-β apresentaram aumentos na expressão entre 12 e 33% no grupo DM. Em IHQ também não houve diferença estatística entre os grupos. O percentual da área do corte histológico marcada para o NF-kB apresentou no grupo NDM uma mediana de 55,09% e nos DM 46,91%, o TGF-β com mediana de 45,46% para os NDM e 56,15% para os DM e o TNF-α com mediana de 54,95% para NDM e 68,88% para DM. A mediana da área do corte histológico marcada para o colágeno foi de 77,90% para NDM e 77,50% para DM, e as fibras elásticas tiveram mediana de 56,52% no grupo de NDM e 69,20% no grupo de DM. Conclusões: A avaliação de marcadores relacionados com a inflamação e estresse oxidativo em fragmentos de artéria torácica interna, não apresentou diferenças ao compararmos pacientes coronariopatas diabéticos versus não-diabéticos. Acreditamos que o pequeno tamanho amostral analisado pode ter contribuído para a ausência de resultados com significância estatística
Introduction: It is known that diabetic patients have a more severe progression of atherosclerotic disease than non-diabetics. Inflammation, oxidative stress and extracellular matrix deposition in the vessel wall are described as key points in the pathophysiology of atherosclerosis, particularly in research made with animal models. There are few studies analyzing these parameters in arteries of medium caliber and diabetic patients compared to non-diabetics. Objectives: To assess the presence of inflammatory markers tumor-α necrosis factor (TNF-α), transformer-β1 growth factor (TGF-β1), nuclear factor kappa B (NF-kB), the enzymes superoxide dismutase (SOD) catalase (CAT) and nitric oxide synthase in inducible form (iNOS) and of the collagen and elastic fibers present in the vascular wall in the graft used in CABG of DM and NDM searching for relationships between results obtained with clinical parameters of the studied groups. Methodology: arterial graft samples from 23 male patients who underwent CABG at the National Institute of Cardiology between December 2014 and December 2015 were included in the study. The evaluation of inflammatory markers expression was performed using relative quantification by real time polymerase chain reaction (qPCR) and immunohistochemistry (IHC). Staining was performed for collagen and elastic fibers in the graft tissue. Results: Clinical data demonstrated that DM and NDM groups were clinically similar with the exception of the existence of diabetes mellitus. The median age of the DM group was 59 years and the NDM group was 64.5 years. Diabetic patients had higher troponin T elevation postoperatively with a median of 5.81 mg / dL while the NDM group had a median of 2.62 mg / dL. The results obtained by qPCR showed no statistical difference among the groups, although there was an increase in CAT expression of 84% in the DM group compared with NDM. The markers NF-kB, SOD and TGF-β showed increases in expression between 12 and 33% in the DM group. In IHC we also did not find statistical difference between the groups. The percentage of the area marked in histologic sections for NF-kB in NDM group showed a median of 55.09% and of 46.91% in DM, for TGF-β a median of 45.46% in NDM and 56.15 % in DM and TNF-α with a median of 54.95% in NDM and 68.88% in DM. The median of the histological section area marked for collagen was 77.90% and 77.50% for NDM and DM, respectively, and elastic fibers had a median of 56.52% in the NDM group and 69.20% in DM group. Conclusions: The evaluation of markers related to inflammation and oxidative stress in internal thoracic artery fragments showed no differences when comparing diabetic coronary patients versus non-diabetics. We believe that the small sample size analyzed may have contributed to the lack of results with statistical significance.
Introduction: It is known that diabetic patients have a more severe progression of atherosclerotic disease than non-diabetics. Inflammation, oxidative stress and extracellular matrix deposition in the vessel wall are described as key points in the pathophysiology of atherosclerosis, particularly in research made with animal models. There are few studies analyzing these parameters in arteries of medium caliber and diabetic patients compared to non-diabetics. Objectives: To assess the presence of inflammatory markers tumor-α necrosis factor (TNF-α), transformer-β1 growth factor (TGF-β1), nuclear factor kappa B (NF-kB), the enzymes superoxide dismutase (SOD) catalase (CAT) and nitric oxide synthase in inducible form (iNOS) and of the collagen and elastic fibers present in the vascular wall in the graft used in CABG of DM and NDM searching for relationships between results obtained with clinical parameters of the studied groups. Methodology: arterial graft samples from 23 male patients who underwent CABG at the National Institute of Cardiology between December 2014 and December 2015 were included in the study. The evaluation of inflammatory markers expression was performed using relative quantification by real time polymerase chain reaction (qPCR) and immunohistochemistry (IHC). Staining was performed for collagen and elastic fibers in the graft tissue. Results: Clinical data demonstrated that DM and NDM groups were clinically similar with the exception of the existence of diabetes mellitus. The median age of the DM group was 59 years and the NDM group was 64.5 years. Diabetic patients had higher troponin T elevation postoperatively with a median of 5.81 mg / dL while the NDM group had a median of 2.62 mg / dL. The results obtained by qPCR showed no statistical difference among the groups, although there was an increase in CAT expression of 84% in the DM group compared with NDM. The markers NF-kB, SOD and TGF-β showed increases in expression between 12 and 33% in the DM group. In IHC we also did not find statistical difference between the groups. The percentage of the area marked in histologic sections for NF-kB in NDM group showed a median of 55.09% and of 46.91% in DM, for TGF-β a median of 45.46% in NDM and 56.15 % in DM and TNF-α with a median of 54.95% in NDM and 68.88% in DM. The median of the histological section area marked for collagen was 77.90% and 77.50% for NDM and DM, respectively, and elastic fibers had a median of 56.52% in the NDM group and 69.20% in DM group. Conclusions: The evaluation of markers related to inflammation and oxidative stress in internal thoracic artery fragments showed no differences when comparing diabetic coronary patients versus non-diabetics. We believe that the small sample size analyzed may have contributed to the lack of results with statistical significance.
Description
Palavras-chave
Aterosclerose, Diabetes mellitus, Enxerto arterial, qPCR, Imunohistoquímica, Inflamação e estresse oxidativo, Atherosclerosis, Arterial graft, Immunohistochemistry, Inflammation and oxidative stress.
Citação
Silva ACR. Análise da expressão de biomarcadores inflamatórios em enxertos utilizados em pacientes diabéticos e não diabéticos submetidos à revascularização do miocárdio. Rio de Janeiro. Tese [Mestrado Profissional em Ciências Cardiovasculares] - Instituto Nacional de Cardiologia; 2016