2型糖尿病阴虚热盛证基因表达的研究

STUDY ON GENE EXPRESSION OF YIN-DEFICIENCY AND EXCESS OF HEAT SYNDROME IN TYPE 2 DIABETES MELLITUS

目的:从分子水平探讨2型糖尿病阴虚热盛证的中医证候实质。方法:抽取2型糖尿病阴虚热盛证患者静脉血,检测血糖、血脂及电解质,分离血液中的白细胞,提取总RNA,化学发光方法检测基因芯片,用ScanAlyze软件采集图像,GEArray Analyzer软件分析数据。对部分差异表达基因进行RT-PCR和Westerm-blotting方法验证。结果:2型糖尿病阴虚热盛证患者血糖、血脂、电解质与健康人有显著差异,基因芯片检测发现和健康人差异表达基因共58条,其中上调的基因有50条,下调的基因有8条。并发现阴虚热盛证的特异性基因。通过RT-PCR和Western-blotting方法反向验证IRS-2和FOXC2基因,证实2型糖尿病阴虚热盛证患者和健康人白细胞中存在IRS-2和FOXC2差异表达。结论:应用基因芯片筛选2型糖尿病阴虚热盛证的差异表达基因,可能为2型糖尿病辨证论治提供新思路。

Objective:To discuss the essence of Chinese medicine syndrome of yin-deficiency and excess of heat in patients with type 2 diabetes mellitus at molecular level.Methods:Collecting venous blood samples of diabetic patients with yin-deficiency and excess of heat syndrome to determinate blood glucose,blood lipid and electrolyte,separate leukocyte and extract total RNA.The gene expression profile was analyzed by scanAlyze and GEArray analyzer software,then the microarray results were confirmed by RT-PCR and Western-blotting.Results:There were significant difference on blood glucose,blood lipid and electrolyte level between type 2 diabetes mellitus patients with yin-deficiency and excess of heat syndrome and healthy people.There were 58 different genes expression of 96 key genes were detected by technique of gene microarray between the two groups,50 genes were up-regulated and 8 genes were down-regulated.The specific genes of yin-deficiency and excess of heat syndrome were found,the differences of FOXC2 and IRS-2 were further confirmed by RT-PCR and Western blotting.Conclusion:There were different gene expression in type 2 diabetes mellitus patients with yin-deficiency and excess of heat syndrome,which can provide valuable information for the differentiation and treatment of diabetes mellitus.