内蒙古农业大学学报（自然科学版）2026，Vol.47Issue(5)：1-7,7.DOI:10.16853/j.cnki.1009-3575.2026.05.001
不同氧气浓度对小鼠肠道精密切片内参基因Ct值的影响
Effect of Different Oxygen Concentrations on the Ct Value of the House Keeping Genes in Mice Precision-cut Intestinal Slices
摘要
Abstract
This study aimed to investigate the impact of different oxygen concentrations(20%and 80%)during culture on the stabili-ty of reference gene Ct values within mouse precision-cut intestinal slices(PCIS).PCIS were prepared and cultured under either 20%O₂(normoxia)or 80%O₂(hyperoxia)for 1,3,6,12,and 24 hours.Total RNA was subsequently extracted,its quality and concentration were assessed,followed by the RT-qPCR analysis to determine the Ct values of the candidate reference genes β-actin,18S rRNA,Ywhaz,Gapdh,and Hprt1.The results showed that,the RNA quality was confirmed to be acceptable(A260/280 absor-bance ratios:1.93-2.03),although the RNA concentration decreased with prolonged culture duration.RT-qPCR analysis revealed that Ywhaz exhibited the highest stability under hyperoxic conditions(80%O₂),with Ct values ranging narrowly from 21.50 to 22.67.β-actin also demonstrated relative stability under hyperoxia(Ct range:15.44-17.74)but exhibited significant variations under normoxic conditions.The genes 18S rRNA,Gapdh,and Hprt1 showed moderate stability under hyperoxia,but their Ct values fluctuated considerably under normoxic conditions.In conclusion,Ywhaz is the most stable reference gene for PCIS cultured under hyperoxic(80%O₂)conditions.While β-actin may also be suitable under hyperoxia,its stability is compromised under normoxia.The stability of 18S rRNA,Gapdh,and Hprt1 is significantly influenced by the ambient oxygen concentration.关键词
精密肠道切片/内参基因/氧气浓度Key words
Precision-cut intestinal slices/Reference genes/Oxygen concentration分类
生物科学引用本文复制引用
云舒,韩旭,贺幼义东,苏日古嘎,哈斯苏荣..不同氧气浓度对小鼠肠道精密切片内参基因Ct值的影响[J].内蒙古农业大学学报（自然科学版）,2026,47(5):1-7,7.基金项目
内蒙古自治区自然科学基金项目(2025QN03082) （2025QN03082）
内蒙古农业大学优秀博士人才引进科研启动项目(NDYB2021-18) （NDYB2021-18）
