Liu Heze, Liu Liang, Han Hui, Liu Kexin, Wang Hui
Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan, 430071, China; Hubei Provincial Key Laboratory of Developmentally Originated Disease, Wuhan, 430071, China.
Department of Orthopedic Surgery, Zhongnan Hospital of Wuhan University, Wuhan, 430071, China.
Reprod Toxicol. 2021 Mar;100:74-82. doi: 10.1016/j.reprotox.2021.01.005. Epub 2021 Jan 14.
The panel of suitable reference genes in the fetal liver have not been reported. In this study, five commonly used reference genes (GAPDH, β-actin, Rn18 s, Rpl13a, and Rps29) were firstly selected as candidates. Bestkeeper, GeNorm, and NormFinder software were then used to screen out the panel of suitable reference genes of male and female fetal rat liver under physiological and prenatal dexamethasone exposure (PDE) conditions. Finally, we verified the reliability of the screened panel of reference genes by standardizing sterol regulatory element binding protein 1c (SREBP1c) expression with different reference genes. The results showed that GAPDH + Rn18 s and GAPDH + Rpl13a were respectively the panel of suitable reference genes in male and female rat fetal liver under the physiological model, while Rn18 s + Rps29 and GAPDH + Rn18 s were respectively under the PDE model. The results showed that different reference genes affected the statistical results of SREBP1c expression, and the screened panel of suitable reference genes under the PDE model had smaller intragroup differences, when compared with other reference genes under physiological and PDE models. In conclusion, we screened and determined that the panel of suitable reference genes were GAPDH + Rn18 s and Rn18 s + Rps29 in the male rat fetal liver under physiological and PDE models, while they were GAPDH + Rpl13a and GAPDH + Rn18 s in the females, and confirmed that the selection of the panel of suitable reference genes in the fetal liver had gender differences and pathological model specificity.
尚未有关于胎儿肝脏中合适的内参基因的报道。在本研究中,首先选择了五个常用的内参基因(甘油醛-3-磷酸脱氢酶(GAPDH)、β-肌动蛋白、18S核糖体RNA(Rn18s)、核糖体蛋白L13a(Rpl13a)和核糖体蛋白S29(Rps29))作为候选基因。然后使用Bestkeeper、GeNorm和NormFinder软件筛选出生理条件下以及产前地塞米松暴露(PDE)条件下雄性和雌性胎鼠肝脏中合适的内参基因组合。最后,我们通过用不同的内参基因标准化固醇调节元件结合蛋白1c(SREBP1c)的表达来验证所筛选的内参基因组合的可靠性。结果表明,在生理模型下,GAPDH + Rn18s和GAPDH + Rpl13a分别是雄性和雌性大鼠胎儿肝脏中合适的内参基因组合,而在PDE模型下,Rn18s + Rps29和GAPDH + Rn18s分别是合适的内参基因组合。结果表明,不同的内参基因会影响SREBP1c表达的统计结果,并且与生理和PDE模型下的其他内参基因相比,PDE模型下所筛选的合适内参基因组合组内差异更小。总之,我们筛选并确定,在生理和PDE模型下,雄性大鼠胎儿肝脏中合适的内参基因组合是GAPDH + Rn18s和Rn18s + Rps29,而雌性大鼠中是GAPDH + Rpl13a和GAPDH + Rn18s,并证实胎儿肝脏中合适的内参基因组合的选择存在性别差异和病理模型特异性。