Abstract: Objective To establish Quantitative Real-time PCR (RT-qPCR) method for detecting the transcription level of xanthine dehydrogenase/oxidase (XDH/XO) gene in Wistar rats，to quantitatively detect XDH/XO gene at the transcriptional level. Method Total RNA was extracted from the liver tissue of Wistar rats. The cDNA was obtained by reverse transcription, and diluted to 5 concentration gradients with 10 dilution factors. RT-qPCR was performed using the designed primer sequences and reference genes to obtain XDH/XO gene standard curve. Furthermore, changes in the transcription level of XDH/XO gene in the animal model of hyperuricemia in Wistar rats were examined. Result The XDH/XO gene standard curve obtained by RT-qPCR method has a single dissolution peak and R2 is close to 1. It can detect the change of XDH/XO gene transcription level in animal model of high uric acid. Conclusion RT-qPCR is a quantitative and accurate method for detecting the XDH/XO gene transcription level in Wistar rats. It can be applied to the pathogenesis of hyperuricemia and new drug research.

Key words: Real-time fluorescence quantitative PCR, XDH/XO, Wistar rats, liver, mRNA

摘要： 目的 建立实时荧光定量(RT-qPCR)检测Wistar大鼠黄嘌呤脱氢酶/氧化酶(XDH/XO)基因转录水平的方法，以在转录水平上对XDH/XO基因进行定量检测。方法 提取Wistar大鼠肝脏组织中总RNA，经逆转录得到cDNA, 以10倍为稀释因子稀释为5个浓度梯度，使用设计的引物序列和内参基因进行RT-qPCR检测，得到XDH/XO基因表达的标准曲线。进而检测Wistar大鼠高尿酸血症动物模型中XDH/XO基因转录水平的变化。结果 RT-qPCR法检测得到的XDH/XO基因标准曲线溶解峰单一，R2接近1，能检测出高尿酸动物模型中XDH/XO基因转录水平的变化。结论 RT-qPCR检测Wistar大鼠XDH/XO基因转录水平的方法具有定量准确，重复性好的特点，可应用于高尿酸血症的发病机理、新药研究等方面。

关键词: 实时荧光定量(RT-qPCR), 嘌呤脱氢酶/氧化酶(XDH/XO), Wistar大鼠, 肝脏, mRNA