Identification of optimal reference genes in golden Syrian hamster with ethanol- and palmitoleic acid-induced acute pancreatitis using quantitative real-time polymerase chain reaction

IF 3.4 Q1 Health Professions Animal models and experimental medicine Pub Date : 2023-05-18 DOI:10.1002/ame2.12321

Jinxin Miao, Le Kang, Tianfeng Lan, Jianyao Wang, Siqing Wu, Yifan Jia, Xia Xue, Haoran Guo, Pengju Wang, Yan Li

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Abstract

Background

Acute pancreatitis (AP) is a severe disorder that leads to high morbidity and mortality. Appropriate reference genes are important for gene analysis in AP. This study sought to study the expression stability of several reference genes in the golden Syrian hamster, a model of AP.

Methods

AP was induced in golden Syrian hamster by intraperitoneal injection of ethanol (1.35 g/kg) and palmitoleic acid (2 mg/kg). The expression of candidate genes, including Actb, Gapdh, Eef2, Ywhaz, Rps18, Hprt1, Tubb, Rpl13a, Nono, and B2m, in hamster pancreas at different time points (1, 3, 6, 9, and 24 h) posttreatment was analyzed using quantitative polymerase chain reaction. The expression stability of these genes was calculated using BestKeeper, Comprehensive Delta CT, NormFinder, and geNorm algorithms and RefFinder software.

Results

Our results show that the expression of these reference genes fluctuated during AP, of which Ywhaz and Gapdh were the most stable genes, whereas Tubb, Eef2, and Actb were the least stable genes. Furthermore, these genes were used to normalize the expression of TNF-α messenger ribonucleic acid in inflamed pancreas.

Conclusions

In conclusion, Ywhaz and Gapdh were suitable reference genes for gene expression analysis in AP induced in Syrian hamster.

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利用实时定量聚合酶链式反应鉴定乙醇和棕榈油酸诱发急性胰腺炎的金色叙利亚仓鼠的最佳参考基因。

背景：急性胰腺炎（AP）是一种导致高发病率和高死亡率的严重疾病。适当的参考基因对于急性胰腺炎的基因分析非常重要。本研究试图研究几种参考基因在金色叙利亚仓鼠（急性胰腺炎的模型）中的表达稳定性：方法：通过腹腔注射乙醇（1.35 克/千克）和棕榈油酸（2 毫克/千克）诱导金色叙利亚仓鼠发生 AP。使用定量聚合酶链反应分析了处理后不同时间点（1、3、6、9 和 24 h）仓鼠胰腺中候选基因（包括 Actb、Gapdh、Eef2、Ywhaz、Rps18、Hprt1、Tubb、Rpl13a、Nono 和 B2m）的表达情况。使用 BestKeeper、Comprehensive Delta CT、NormFinder 和 geNorm 算法和 RefFinder 软件计算了这些基因的表达稳定性：结果表明，这些参考基因的表达在AP期间出现波动，其中Ywhaz和Gapdh是最稳定的基因，而Tubb、Eef2和Actb则是最不稳定的基因。此外，这些基因还被用来归一化炎症胰腺中TNF-α信使核糖核酸的表达：总之，Ywhaz和Gapdh是叙利亚仓鼠AP诱导基因表达分析的合适参考基因。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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