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目的:建立碳水化合物反应元件结合蛋白(ChREBP)基因表达的绝对定量实时PCR检测技术。方法:32只SD大鼠随机分为常氧安静组、常氧运动组、低氧安静组和低氧运动组,运动组每天运动1小时,每周运动5天;低氧组每天在低氧下生活10小时。实验6周后取肝组织,应用实时定量PCR方法检测肝组织ChREBP mRNA的表达。结果:实时定量PCR方法可对肝组织ChREBP mRNA表达进行定量检测,ChREBP基因拷贝数与检测阈值呈线性关系,相关系数r>0.99。结论:实时定量PCR方法检测ChREBP mRNA结果准确、可靠。
Objective: To establish an absolute quantitative real-time PCR assay for the expression of ChREBP gene. Methods: Thirty-two Sprague-Dawley rats were randomly divided into normoxia group, normoxia group, hypoxia group and hypoxia group. The exercise group exercised for 1 hour a day and exercised for 5 days a week. In the hypoxia group, Life 10 hours. Six weeks after the experiment, liver tissue was taken and the expression of ChREBP mRNA in liver tissue was detected by real-time quantitative PCR. Results: Quantitative detection of ChREBP mRNA in liver tissue by quantitative real-time PCR showed that the ChREBP gene copy number was linearly correlated with the detection threshold, and the correlation coefficient was> 0.99. Conclusion: The real-time quantitative PCR method to detect ChREBP mRNA results is accurate and reliable.