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

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