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鉴定注射β酪啡肽的 C57BL/6 小鼠肝脏中表达数据正常化的稳定内参基因（ICGs）。

Identification of stably expressed Internal Control Genes (ICGs) for normalization of expression data in liver of C57BL/6 mice injected with beta casomorphins.

机构信息

Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana, India.

Animal Biotechnology Division, National Bureau of Animal Genetic Resources, Karnal, Haryana, India.

出版信息

PLoS One. 2023 May 5;18(5):e0282994. doi: 10.1371/journal.pone.0282994. eCollection 2023.

DOI:10.1371/journal.pone.0282994
PMID:37145997
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10162558/
Abstract

In recent years, beta-casomorphin peptides (BCM7/BCM9) derived from the digestion of cow milk have drawn a lot of attention world over because of their proposed impact on human health. In order to evaluate the transcriptional modulation of target genes through RT-qPCR in response to these peptides, availability of appropriate reference or internal control genes (ICGs) will be the key. The present study was planned to identify a panel of stable ICGs in the liver tissue of C57BL/6 mice injected with BCM7/BCM9 cow milk peptides for 3 weeks. A total of ten candidate genes were evaluated as potential ICGs by assessing their expression stability using software suites; geNorm, NormFinder and BestKeeper. The suitability of the identified ICGs was validated by assessing the relative expression levels of target genes, HP and Cu/Zn SOD. Based on geNorm, PPIA and SDHA gene pair was identified to be most stably expressed in liver tissue during the animal trials. Similarly, NormFinder analysis also identified PPIA as the most stable gene. BestKeeper analysis showed crossing point SD value for all the genes in the acceptable range that is closer to 1. Overall, the study identified a panel of stable ICGs for reliable normalization of target genes expression data in mice liver tissues during BCM7/9 peptides trial.

摘要

近年来，源于牛奶消化的β-酪啡肽（BCM7/BCM9）因其对人类健康的潜在影响而备受关注。为了通过 RT-qPCR 评估这些肽对靶基因的转录调控，合适的参考或内参基因（ICGs）的可用性将是关键。本研究旨在鉴定一组在注射 BCM7/BCM9 牛奶肽 3 周的 C57BL/6 小鼠肝脏组织中稳定的 ICGs。通过使用软件套件 geNorm、NormFinder 和 BestKeeper 评估其表达稳定性，共评估了 10 个候选基因作为潜在的 ICGs。通过评估靶基因 HP 和 Cu/Zn SOD 的相对表达水平来验证鉴定的 ICGs 的适用性。基于 geNorm，鉴定出 PPIA 和 SDHA 基因对在动物试验中在肝脏组织中表达最稳定。同样，NormFinder 分析也鉴定出 PPIA 是最稳定的基因。BestKeeper 分析显示所有基因的 CP 值 SD 值均在可接受范围内，更接近 1。总体而言，该研究鉴定了一组稳定的 ICGs，可用于在 BCM7/9 肽试验期间可靠地对小鼠肝脏组织中靶基因表达数据进行归一化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/785b/10162558/4d029f46d015/pone.0282994.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/785b/10162558/e3dc4ebc941e/pone.0282994.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/785b/10162558/201292357b60/pone.0282994.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/785b/10162558/065b104a5795/pone.0282994.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/785b/10162558/36023c2e2ab0/pone.0282994.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/785b/10162558/b262ffd4570a/pone.0282994.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/785b/10162558/4d029f46d015/pone.0282994.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/785b/10162558/e3dc4ebc941e/pone.0282994.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/785b/10162558/201292357b60/pone.0282994.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/785b/10162558/065b104a5795/pone.0282994.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/785b/10162558/36023c2e2ab0/pone.0282994.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/785b/10162558/b262ffd4570a/pone.0282994.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/785b/10162558/4d029f46d015/pone.0282994.g006.jpg

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