应用基因表达谱芯片技术筛选老龄肾阳虚证特征功能类基因

Screening and Identification of Functional Gene Sets in Aged Kidney-Yang Deficiency Syndrome:cDNA microarray assay

目的:利用基因芯片技术探讨肾阳虚证发生的特征功能基因集。方法:在老龄肾阳虚证流行病学调查过程中,筛选出典型肾阳虚证患者2例和正常对照2例,采用美国Affymetrix公司的HG-u133Plus2.0基因表达谱芯片,按一步法抽提总RNA,分离纯化两种样品的mRNA,经逆转录合成掺入生物素标记的cDNA合成探针,与芯片杂交和严格洗片后,用荧光扫描仪扫描芯片荧光信号图像,获得肾阳虚患者差异表达基因。结果:两例患者和正常人比较获得680和503条差异表达基因,通过g:GOSt基因芯片数据分析平台进行基因功能分类,发现免疫反应功能类基因出现显著性。结论:肾阳虚证的发生与免疫反应功能类基因的异常表达密切相关。

Objective：To screen and identify functional gene sets in aged kidney-Yang deficiency syndrome by eDNA microarray assay. Methods ： Two aged kidney-Yang deficiency patients and two healthy peoples were selected as samples during the epidemialogic survey of the kidney-Yang deficiency syndrome. The differentially expressed genes were screened by applying HGu133Plus2. 0 of Affymetrix GeneChip. The total RNAs were isolated from leukocyte of 4 samples by TR Izolmethod and then purified, reversely transcribed to eDNA with incorporating biotin labeling probe and hybridizated with GeneChip. Picture signala of fluorescence in gene array were scanned and compared differential expression of genes by computer analysis. Results：680 and 503 differentially expressed genes were obtained. The immune response-associated gene sets were found with significance in GO annotated biological processes. Conclusion：Immune factor play an important role in occurrence and development of the kidney-Yang deficiency.