中医寒体与热体基因差异性表达的RT-PCR分析

Analysis of RT-PCR on Difference of Gene Expression in Rats of Cold Body and Heat Body Constitution

目的:用基因芯片技术筛选出寒体与热体的特征性基因,并对基因差异性表达进行RT-PCR分析。方法:通过测量大鼠两后脚掌掌心温度,将其分为寒体组、热体组和常体组,取各组大鼠的肝脏组织,抽提总RNA来进行逆转录,标记后与大鼠全基因组芯片杂交,分析3组基因表达谱的差异情况,并用RT-PCR技术对其中部分表达差异基因的表达水平变化进行验证。结果:基因Atp6n l、Ins2、AChE、Txnrd1在热体组和寒体组中的表达存在差异,且具有统计学意义(P<0.05)。结论:寒体和热体在基因水平和基因表达水平上存在显著差异,可为临床通过药物或食物调节特异性基因表达水平来改善体质提供了前提条件。

Objective ： To apply the gene chip technology to select the characteristic genes in rats of cold body and heat body constitution, and analyze the differencc of gene expression by RT-PCR. Methods： The rats were divided into cold body group, hot body group and normal body group by measuring the temperatures of two posterior soles. The liver tissues of all rats were obtained, and the total RNA was extracted for reverse transcription. After labeling, the probes were hybridized with rat genome array, the differences of gene expressions in three groups were analyzed, and the levels of expressions of different genes were detected by RT-PCR technology. Results ： There were statistical differences on the expressions of Atp6nl, Ins2, AChE and Txnrdl gene between heat body group and cold body group （ P 〈9.0S）. Conclusion： There are differences on the level of gene expression between cold body and heat body, and it provides a prerequisite to regulating characteristic gene expression through drug or food in clinic for improving constitution.