Araujo Layanne C C, Bordin Silvana, Carvalho Carla R O
Department of Physiology and Biophysics, Institute of Biological Science, University of Sao Paulo, Sao Paulo 05508-900, Brazil.
Gastroenterol Res Pract. 2020 Feb 3;2020:1093235. doi: 10.1155/2020/1093235. eCollection 2020.
The expression levels of some reference genes and proteins are used for data normalization and quantification. However, these levels can change in response to experimental conditions or treatments. . The aim of this work was to evaluate reference gene and protein expression in models of nonalcoholic fatty liver disease, using mice fed with a high-fat diet (HFD) and mice that are genetically obese (ob/ob). . Histological staining techniques were used to verify the morphology and quantify the amount of lipid droplets present in the liver. Real-time polymerase chain reaction and immunoblotting were employed for monitoring protein expression and gene expression levels, respectively. . The results showed that there was a substantial increase in the amount of lipid droplets in the livers of HFD and ob/ob animals when compared to the standard diet (SD) group. There was an observed reduction in the expression of -actin (10%), -tubulin (6%), GAPDH (19%), and RPL3 (15%) genes when comparing the ob/ob group to the HFD group. Additionally, the ob/ob mice displayed GAPDH protein levels that were substantially, but not significantly, reduced when compared to SD. . It was concluded that there are slight differences in the expression levels of reference genes and proteins in these two NAFLD animal models, and researchers should consider these alterations when working with these models.
一些参考基因和蛋白质的表达水平用于数据标准化和定量。然而,这些水平会因实验条件或处理而发生变化。本研究的目的是评估非酒精性脂肪性肝病模型中参考基因和蛋白质的表达,使用高脂饮食(HFD)喂养的小鼠和遗传性肥胖(ob/ob)小鼠。组织学染色技术用于验证肝脏的形态并量化其中存在的脂滴数量。实时聚合酶链反应和免疫印迹分别用于监测蛋白质表达和基因表达水平。结果表明,与标准饮食(SD)组相比,HFD和ob/ob动物肝脏中的脂滴数量大幅增加。将ob/ob组与HFD组进行比较时,观察到β-肌动蛋白(10%)、β-微管蛋白(6%)、甘油醛-3-磷酸脱氢酶(GAPDH,19%)和核糖体蛋白L3(RPL3,15%)基因的表达降低。此外,与SD相比,ob/ob小鼠的GAPDH蛋白水平大幅降低,但无统计学意义。得出的结论是,在这两种非酒精性脂肪性肝病动物模型中,参考基因和蛋白质的表达水平存在细微差异,研究人员在使用这些模型时应考虑这些变化。