基因芯片技术筛选2型糖尿病胰岛素抵抗脂肪代谢相关差异表达基因

目的:应用基因芯片技术比较2型糖尿病胰岛素抵抗(IR)患者和正常人大网膜组织脂肪代谢相关基因表达谱的差异,探讨IR可能的发病机制.方法:分别用Cy5 和Cy3两种不同的荧光染料通过逆转录反应将IR组(n＝5)和正常对照组(n＝5)脂肪组织的mRNA标记成探针,并与载有一组靶基因的基因表达谱芯片进行杂交,通过扫描荧光强度,计算机软件分析,寻找两组差异表达基因.然后对芯片筛选结果中差异表达的FOXC2基因用Northern印迹法进行验证.结果:IR组和正常对照组之间共筛选出82条差异表达已知基因;其中脂肪代谢相关基因10条,表达增加的基因5条,表达降低的基因5条.Northern印迹证实IR组FOXC2 mRNA表达明显升高,与基因芯片检测结果一致. 结论:IR的发病与脂肪代谢相关,FOXC2可能是2型糖尿病IR的候选基因.

cDNA microarray in screening of differentially expressed genes associated with lipid metabolism in adipose tissues of patients with insulin resistant type 2 diabetes mellitus

Objective:To study the differentially expressed genes associated with lipid metabolism in adipose tissues of patients with insulin resistant(IR) type 2 diabetes mellitus(T2DM),in an effort to explore the mechanism of IR.Methods: mRNA from the omental adipose tissues of T2DM patients(n=5) and normal controls(n=5) were reversely transcribed into cDNAs with the incorporation of fluorescent dUTP(cy-5 or cy-3) to prepare hybridization probes.The mixed probes were hybridized with a cDNA microarray containing the target genes.The results were scanned and subjected to computer analysis to search for the difference between the gene spectrum of T2DM patients and normal controls.The differentially expressed gene,FOXC2,was verified by Northern blot.Results: Eighty-two differentially expressed genes were identified between these 2 groups,with 10 associated with lipid metabolism,including 5 upregulated ones and 5 downregulated ones.Northern blotting confirmed that the expression of FOXC2 mRNA was increased in IR group,which was in accordance with the result of cDNA microarray analysis.Conclusion: The pathogenesis of IR is related with lipid metabolism and the FOXC2 gene may be a candidate gene of(IR. )