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目的观察12个常用看家基因在正常小鼠和哮喘小鼠颈静脉-结状神经节的表达变化,选择合适的内参基因为进一步基因表达分析研究得到可信数据提供依据。方法雌性BALB/c小鼠随机分为哮喘模型组和对照组,制备哮喘模型,用实时荧光定量PCR技术,经geNorm及Normfinder程序分析,评价了12个常用看家基因CYC1、UBC、RPL13、YWHAZ、18srRNA、B2M、SDHA、ATP5B、CANX、ACTB、EIF4A2和GAPDH在小鼠颈-结状神经节的表达稳定性。结果 12个候选看家基因在小鼠神经节的表达差异较大,其中GAPDH、18srRNA、CANX、ACTB和EIF4A2表达相对稳定,而CYC1、UBC和RPL13的表达在两组小鼠神经节变化幅度较大,结合两种分析证实GAPDH是其中最稳定的内参基因。结论本研究结果提示GAPDH是研究哮喘小鼠和正常小鼠颈静脉-结状神经节基因表达的最佳内参基因,同时也为进一步认识在不同实验模型确定适宜内参基因的重要性提供依据。
Objective To observe the expression of 12 common housekeeping genes in the jugular vein-nodose ganglion of normal mice and asthmatic mice, and to select suitable internal reference genes for further gene expression analysis to provide credible data. Methods Female BALB / c mice were randomly divided into asthma model group and control group, asthma model was prepared, real-time fluorescent quantitative PCR technology, geNorm and Normfinder program analysis, evaluation of 12 commonly used housekeeping genes CYC1, UBC, RPL13, YWHAZ , 18srRNA, B2M, SDHA, ATP5B, CANX, ACTB, EIF4A2 and GAPDH in mouse cervical nodose ganglia. Results The expression of 12 candidate housekeeping genes was significantly different in the mouse ganglia. The expression of GAPDH, 18srRNA, CANX, ACTB and EIF4A2 was relatively stable, while the expressions of CYC1, UBC and RPL13 in the ganglion were significantly different Large, combined with two kinds of analysis confirmed that GAPDH is one of the most stable internal control gene. Conclusions The results of this study suggest that GAPDH is the best internal control gene for studying the gene expression of jugular vein-nodose ganglion in asthmatic mice and normal mice. It also provides a basis for further understanding the importance of determining appropriate internal control genes in different experimental models.