Jiang Congshan, Meng Liesu, Zhu Wenhua, Shahzad Muhammad, Yang Xudong, Lu Shemin
Department of Genetics and Molecular Biology, Xi'an Jiaotong University School of Medicine, Post Box 81, Western Yanda Road 76, Xi'an, Shaanxi, 710061, People's Republic of China.
Inflamm Res. 2009 Sep;58(9):601-9. doi: 10.1007/s00011-009-0027-5. Epub 2009 Apr 9.
Aim of this study is to explore the expression stability of 12 commonly used housekeeping genes in inflammation condition.
Two rat models, pristane-induced arthritis (PIA) and antigen-induced pulmonary inflammation (AIPI) were established and reverse transcription-PCR was performed from rat spleens and inguinal lymph nodes.
The reference gene expression fluctuated in the immune-associated tissues and 18S rRNA expression from the various inflammatory conditions was the most stably in all examined gene expression. Besides, peptidylprolyl isomerase A and beta-glucuronidase in PIA spleens, beta-actin and GAPDH in PIA inguinal lymph nodes, and peptidylprolyl isomerase A and beta-actin in AIPI spleens, expressed more stably than others. These results were further confirmed by expression study of TNF-alpha in PIA rat inguinal lymph nodes.
We suggest that the stable gene mentioned above or geometric mean of three best references in each experimental condition should be used as the reference and the variable ones should be avoided in similar study designs.
本研究旨在探讨12种常用管家基因在炎症状态下的表达稳定性。
建立了两种大鼠模型，即 pristane诱导的关节炎（PIA）和抗原诱导的肺部炎症（AIPI），并从大鼠脾脏和腹股沟淋巴结进行逆转录-聚合酶链反应。
参考基因在免疫相关组织中的表达存在波动，在所有检测的基因表达中，来自各种炎症状态的18S rRNA表达最稳定。此外，PIA脾脏中的肽基脯氨酰异构酶A和β-葡萄糖醛酸酶、PIA腹股沟淋巴结中的β-肌动蛋白和甘油醛-3-磷酸脱氢酶以及AIPI脾脏中的肽基脯氨酰异构酶A和β-肌动蛋白，表达比其他基因更稳定。这些结果在PIA大鼠腹股沟淋巴结中TNF-α的表达研究中得到了进一步证实。
我们建议在类似的研究设计中，应使用上述稳定基因或每种实验条件下三个最佳参考基因的几何平均值作为参考，避免使用不稳定的基因。
