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在体外非酒精性脂肪肝疾病中实时聚合酶连锁反应的定量参考基因

Jingzhi Chen¹, Xingmi Chen², Ying Liu²

¹School of Integrative Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, China.

|December 3, 2025

概括

这项研究确定Ywhaz是自由脂肪酸 (FFA) 诱导的代谢功能障碍相关脂肪肝疾病 (MAFLD) 模型中定量PCR (qPCR) 最稳定的参考基因. 此外,还建立了一个可靠的选程序.

科学领域:

生物化学生化学
细胞生物学细胞生物学
分子生物学分子生物学

背景情况:

参考基因是必不可少的,但在代谢功能障碍相关的脂肪肝疾病 (MAFLD) 研究中往往没有得到验证.
通过定量PCR (qPCR) 准确的基因表达分析依赖于稳定的参考基因.

研究的目的:

在自由脂肪酸 (FFA) 诱导的MAFLD细胞模型中识别和验证qPCR最稳定的参考基因.
建立一种可靠的方法来选MAFLD研究中的参考基因.

主要方法:

RNA测序 (RNA-seq) 在对照和FFA治疗的AML12细胞上进行.
候选参考基因从文献中选择,并使用RNA-seq数据进行过 (双指数排名,变化系数,Log2FoldChange).
用多个算法 (NormFinder,BestKeeper,geNorm,ΔCt,RefFinder) 评估候选基因的表达稳定性.

主要成果:

FFA治疗导致AML12细胞中显著的脂质滴积累.
RNA-seq分析和RefFinder确定Ywhaz是最稳定的参考基因.
与其他候选人相比,Ywhaz表现出优越的表达稳定性.

结论:

使用AML12细胞模型,Ywhaz是使用FFA诱导的MAFLD中的qPCR最稳定的参考基因.
该研究提供了一种验证的程序,用于在MAFLD研究中选择参考基因.
这一发现支持在MAFLD中进行更准确的基因表达研究.

关键词:

马夫尔德 (MAFLD) 是一个有关RNA测序的RNA测序参考基因是一种基因.qPCRR 是一个很好的方法.

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