2型糖尿病中医湿热困脾证的基因表达研究

Study of Gene Expression of Damp-heat-accumulate Spleen with Type 2 Diabetes Mellitus

[目的]从分子水平探讨2型糖尿病湿热困脾证的中医证候实质。[方法]抽取2型糖尿病气阴两虚证患者静脉血,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 Ana-lyzer软件分析数据。对部分差异表达基因进行RT-PCR和Western-blotting方法验证。[结果]2型糖尿病湿热困脾证患者血糖,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38条,其中上调的基因有24条,下调的基因有14条。并发现湿热困脾证的特异性基因。通过RT-PCR和Western-blotting方法反向验证IRS-2和FOXC2基因,证实2型糖尿病湿热困脾证患者和健康人白细胞中存在IRS-2和FOXC2存在差异表达。[结论]应用基因芯片筛选2型糖尿病湿热困脾证的差异表达基因,为2型糖尿病辨证论治提供新思路。

[Objective]Study type 2 diabetes mellitus of deficiency of damp-heat-accumulate spleen of syndromes at molecular level.[Methods] Hemospasia venous blood samples of diabetes,measured bloodg lucose,blood lipid and electroyte.Extracti leukocyte and RNA.The gene expression profile was analyzed by scan Alyze and gEArray Analyzer software.And then the microarray results were confirmed by RT-PCR and Western-blotting.[Results]The results of bloodglucose,blood lipid and electroyte had significant differences between diabetes and healthy subjects.Use Diabetesgene Array of Superarray Bioscience,containing 96 key genes,find 38 different expressiongene,24 up-regulatedgenes and 14 down-regulatedgenes,and find the specific gene of damp-heat-accumulate spleen of syndromes;FOXC2,IRS-2 mRNA were further confirmed by RT-PCR and IRS-2 protein confirmed by western-blotting respectively and the expression of them was coincided well with the gene chip results.[Conclusion]The screen of gene expression in type 2 diabetes mellitus of damp-heat-accumulate spleen of syndromes can provide valuable information for the research of diabetes mellitus syndrome differentiation and treatments.