小鼠哮喘模型颈静脉-结状神经节内参基因的筛选

Selection of reference genes for expression studies in the jugular-nodose ganglion of a mouse model of asthma

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 investigate the expression stability of 12 common housekeeping genes （HKGs） in jugular-nodose ganglion complex （JNC） from control and asthmatic mice. Methods Female BALB/e mice were randomly divided into control group and asthma group. Expression levels of 12 candidate HKGs, CYC1, UBC, RPL13, YWHAZ, 18srRNA, B2M, SDHA, ATP5B, CANX, ACTB, EIF4A2 and GAPDH, were measured by quantitative real-time PCR in JNCs from 5 normal mice and 5 asthmatic mice. Gene expression stability was subsequently determined using geNorm and Normfinder applets. Results GAPDH, 18srRNA, CANX, ACTB and EIF4A2 were relative stably expressed. However, the expression levels of CYC1, UBC and RPL13 were significantly different between the two groups. GAPDH was ranked the most stable by geNorm and Normfmder. Conclusions The results suggest that GAPDH is the most suitable housekeeping gene analyzed for expression studies in JNCs from normal and asthmatic mice, and provide the evidence of the importance of validating housekeeping genes for different experimental model.