During quantitative real-time polymerase chain reaction (RT-qPCR) data analysis, the selection of optimal housekeeping gene is necessary to ensure the accuracy of results. It is noteworthy that housekeeping genes commonly used in adult studies may not be applicable for fetus. However, the stability analysis of housekeeping gene in fetal kidney has not been reported. This study intends to screen the applicable compound housekeeping genes in rat fetal kidney. In this study, eight housekeeping genes used in kidney studies based on literature reports (GAPDH, ACTB, 18S, HPRT, YWHAZ, HMBS, PPIA, and TBP) were selected as the research object. Their expression levels in the rat fetal kidney in physiological condition and the intrauterine growth retardation (IUGR) model induced by prenatal dexamethasone exposure (PDE) (0.2 mg/kg·day from gestation Days 9 to 20) was measured. Furthermore, these eight housekeeping genes were used to conduct relative quantitative analysis of nephrin expression in the fetal kidney in PDE-induced IUGR model, to compare the influence of choosing different housekeeping gene on data analysis of nephrin expression and to verify the reliability of selected compound housekeeping genes. In this study, stable housekeeping genes of fetal kidney tissues in PDE-induced IUGR model were identified: ACTB, GAPDH, TBP, and HMBS for males; ACTB, YWHAZ, and GAPDH for females. Besides, our results suggest that ACTB + GAPDH were the best compound housekeeping genes for normalization analysis in male fetal kidney studies, and ACTB + YWHAZ in females. This study will provide an experimental evidence basis for the selection of housekeeping genes in the RT-qPCR experiment in renal development toxicology-related models.

中文翻译：

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用于大鼠胎肾定量实时聚合酶链反应分析的复合看家基因的选择和鉴定

在定量实时聚合酶链反应 (RT-qPCR) 数据分析过程中，需要选择最佳管家基因以确保结果的准确性。值得注意的是，成人研究中常用的管家基因可能不适用于胎儿。然而，关于管家基因在胎儿肾脏中的稳定性分析尚未见报道。本研究旨在筛选大鼠胎肾中适用的复合看家基因。在这项研究中，基于文献报道的肾脏研究中使用了八种管家基因（GAPDH、ACTB、18S、HPRT、YWHAZ、HMBS、PPIA和TBP) 被选为研究对象。测定了它们在生理条件下大鼠胎肾和产前地塞米松暴露（PDE）（0.2 mg/kg·day，妊娠第9-20天）诱导的宫内发育迟缓（IUGR）模型中的表达水平。此外，将这8个看家基因用于PDE诱导的IUGR模型中胎儿肾脏nephrin表达的相对定量分析，比较选择不同看家基因对nephrin表达数据分析的影响，并验证所选化合物的可靠性。管家基因。本研究鉴定了PDE诱导的IUGR模型中胎儿肾组织的稳定管家基因：ACTB、GAPDH、男性的TBP和HMBS；女性的ACTB、YWHAZ和GAPDH。此外，我们的结果表明，在男性胎儿肾脏研究中， ACTB  +  GAPDH是用于标准化分析的最佳复合管家基因，而在女性中，ACTB  + Y WHAZ是最佳复合管家基因。本研究将为肾发育毒理学相关模型中RT-qPCR实验中管家基因的选择提供实验依据。