The Nile rat (Arvicanthis niloticus) is a novel diurnal carbohydrate-sensitive rodent useful for studies on type 2 diabetes mellitus (T2DM) and the metabolic syndrome. Hepatic responses to T2DM and any interventions thereof can be evaluated via transcriptomic gene expression analysis. However, the study of gene expression via real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) requires identification of stably expressed reference genes for accurate normalisation. This study describes the evaluation and identification of stable reference genes in the livers from Control Nile rats as well as those supplemented with Water-Soluble Palm Fruit Extract, which has been previously shown to attenuate T2DM in this animal model. Seven genes identified as having stable expression in RNA-Sequencing transcriptome analysis were chosen for verification using real-time RT-qPCR. Six commonly used reference genes from previous literature and two genes from a previous microarray gene expression study in Nile rats were also evaluated. The expression data of these 15 candidate reference genes were analysed using the RefFinder software which incorporated analyses performed by various algorithms. The Hpd, Pnpla6 and Vpp2 genes were identified as the most stable across the 36 samples tested. Their applicability was demonstrated through the normalisation of the gene expression profiles of two target genes, Cela1 and Lepr. In conclusion, three novel reference genes which can be used for robust normalisation of real-time RT-qPCR data were identified, thereby facilitating future hepatic gene expression studies in the Nile rat.

尼罗河大鼠（Arvicanthis niloticus）是一种新型的昼间碳水化合物敏感型啮齿动物，可用于研究2型糖尿病（T2DM）和代谢综合征。可以通过转录组基因表达分析评估对T2DM的肝反应及其任何干预。但是，通过实时逆转录定量聚合酶链反应（RT-qPCR）进行基因表达的研究需要鉴定稳定表达的参考基因，以进行准确的标准化。这项研究描述了来自对照尼罗河大鼠以及补充有水溶性棕榈果提取物的肝脏中稳定的参考基因的评估和鉴定，该参考基因先前已在该动物模型中减弱了T2DM。选择了在RNA测序转录组分析中具有稳定表达的七个基因，用于使用实时RT-qPCR进行验证。还评估了来自尼罗河大鼠先前文献中的六个常用参考基因​​和来自先前微阵列基因表达研究中的两个基因。使用RefFinder软件分析了这15个候选参考基因的表达数据，该软件结合了由各种算法执行的分析。的在测试的36个样本中，Hpd，Pnpla6和Vpp2基因被认为是最稳定的。通过对两个靶基因Cela1和Lepr的基因表达谱进行归一化，证明了它们的适用性。总之，鉴定了三个可用于实时RT-qPCR数据稳健归一化的新参考基因，从而促进了今后在尼罗河大鼠中肝基因表达的研究。