Bronchopulmonary dysplasia is a chronic lung disease of preterm infants. Mouse models of hyperoxia-induced lung injury are often used to study pathogenesis and potential therapeutic approaches of BPD. Beside histological studies, gene expression analysis of lung tissue is typically used as experimental read out. RT-qPCR is the standard method for gene expression analysis, however, the accuracy of the quantitative data depends on the appropriate selection of reference genes. No data on validated reference genes for hyperoxia-induced neonatal lung injury in mice is available. In this study, 12 potential reference genes were systematically analyzed for their expression stability in lung tissue of neonatal mice exposed to room air or hyperoxia and healthy adult controls using published software algorithms. Analysis of gene expression data identified Hprt, Tbp and Hmbs as the most stable reference genes and proposed combinations of Hprt/Sdha or Hprt/Rpl13a as potential normalization factors. These reference genes and normalization factors were validated by comparing Il6 gene and protein expression and may facilitate accurate gene expression analysis in lung tissues of similar designed studies.

中文翻译：

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在基于高氧的支气管肺发育不良小鼠模型中进行基因表达分析的候选参考基因的系统分析

支气管肺发育不良是早产儿的一种慢性肺部疾病。高氧诱导肺损伤的小鼠模型常用于研究 BPD 的发病机制和潜在的治疗方法。除了组织学研究，肺组织的基因表达分析通常用作实验读数。RT-qPCR is the standard method for gene expression analysis, however, the accuracy of the quantitative data depends on the appropriate selection of reference genes. 没有关于小鼠高氧诱导的新生儿肺损伤的经过验证的参考基因的数据。在这项研究中，使用公开的软件算法系统地分析了 12 个潜在参考基因在暴露于室内空气或高氧的新生小鼠和健康成人对照的肺组织中的表达稳定性。基因表达数据分析确定 Hprt、Tbp 和 Hmbs 是最稳定的参考基因，并建议将 Hprt/Sdha 或 Hprt/Rpl13a 组合作为潜在的标准化因子。通过比较 Il6 基因和蛋白质表达来验证这些参考基因和标准化因子，并且可能有助于类似设计研究的肺组织中的准确基因表达分析。