Analysis of the stability of housekeeping gene expression in the left cardiac ventricle of rats submitted to chronic intermittent hypoxia
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The effects of chronic intermittent hypoxia on housekeeping gene expression in the left cardiac ventricle of rats is analyzed and all genes were found to be suitable for use, glyceraldehyde-3-phosphate dehydrogenase and 18S being classified as the most and the least stable, respectively.Abstract:
Obstructive sleep apnea (OSA) has been associated with oxidative stress and various cardiovascular consequences, such as increased cardiovascular disease risk. Quantitative real-time PCR is frequently employed to assess changes in gene expression in experimental models. In this study, we analyzed the effects of chronic intermittent hypoxia (an experimental model of OSA) on housekeeping gene expression in the left cardiac ventricle of rats. Analyses via four different approaches-use of the geNorm, BestKeeper, and NormFinder algorithms; and 2-ΔCt (threshold cycle) data analysis-produced similar results: all genes were found to be suitable for use, glyceraldehyde-3-phosphate dehydrogenase and 18S being classified as the most and the least stable, respectively. The use of more than one housekeeping gene is strongly advised. RESUMO A apneia obstrutiva do sono (AOS) tem sido associada ao estresse oxidativo e a varias consequencias cardiovasculares, tais como risco aumentado de doenca cardiovascular. A PCR quantitativa em tempo real e frequentemente empregada para avaliar alteracoes na expressao genica em modelos experimentais. Neste estudo, analisamos os efeitos da hipoxia intermitente cronica (um modelo experimental de AOS) na expressao de genes de referencia no ventriculo cardiaco esquerdo de ratos. Analises a partir de quatro abordagens - uso dos algoritmos geNorm, BestKeeper e NormFinder e analise de dados 2-ΔCt (ciclo limiar) - produziram resultados semelhantes: todos os genes mostraram-se adequados para uso, sendo que gliceraldeido-3-fosfato desidrogenase e 18S foram classificados como o mais e o menos estavel, respectivamente. A utilizacao de mais de um gene de referencia e altamente recomendada.read more
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