史晓贤 李高文

[摘要] 目的 探讨基因表达谱在筛选糖尿病肾病有效治疗药物中的应用效果。 方法 在GEO数据库中搜索与糖尿病肾病相关的基因芯片，利用d-chip软件进行差异基因的表达分析，采用基因集富集的方法（GSEA）进行相关信号通路的富集分析，再利用Cmap筛选治疗糖尿病肾病的候选药物，并通过动物实验进行药效学评估。 结果 通过对GSE31022芯片中的合格样本进行差异基因的有效分析，在获得的540个差异表达基因中，上调基因150个，下调基因390个；通过GSEA分析主要富集到NF-κB信号通路与糖尿病肾病最为密切（P < 0.05），并且筛选出甲砜霉素可能对糖尿病肾病有治疗作用（score=1），动物实验结果显示，与模型组比较，甲砜霉素组的肾小球硬化及系膜扩张明显改善，基本达到正常组水平，差异有统计学意义（P < 0.05）。此外，甲砜霉素组的尿蛋白和CD68、转化生长因子β、纤溶酶原激活物抑制物1及白介素6的mRNA水平低于模型组（P < 0.05）。 结论 基于糖尿病肾病相关基因表达谱作为疾病有效治疗药物的预测手段，为疾病的治疗药物发现提供了新思路。

[关键词] 基因芯片；糖尿病肾病；甲砜霉素；基因表达谱；富集分析

[中图分类号] R587.2 [文献标识码] A [文章编号] 1673-7210（2018）08（a）-0018-05

[Abstract] Objective To explore the application effects of gene expression profiling in screening the effective therapeutic drugs for diabetic nephropathy. Methods The gene chips related to diabetic nephropathy in GEO databases were searched and the differential gene expression was analyzed by d-chip software. Enrichment analysis of the relevant signaling pathways was carried out by GSEA， Cmap was then used for the screening of candidate drugs of diabetic nephropathy and pharmacodynamic validation was evaluated in animal experiments. Results Through the analysis of differential genes in qualified samples from the GSE31022 chip， a total of 540 genes were obtained， of which， 150 genes were up-regulated and 390 genes were down-regulated. The main enrichment to NF-κB signaling pathway by GSEA was most closely related to diabetic nephropathy （P < 0.05）， and screening for Thiamphenicol may have a therapeutic effect on diabetic nephropathy （score=1）. The results of animal experiments showed that compared with model group， the mesangial expansion of the diabetic nephropathy mice treated by Thiamphenicol was significantly improved and almost reached the normal level （P < 0.05）. In addition， the levels of urine protein and the mRNA levels of CD68， transforming growth factor β， plasminogen activator inhibitor 1 and interleukin 6 in the Thiamphenicol group were significantly lower than those in model group （P < 0.05）. Conclusion As an effective means of predicting effective therapeutic drugs for diabetic nephropathy， the gene expression profiling based on diabetic nephropathy provides a new idea for the discovery of therapeutic drugs for the disease.

[Key words] Gene chips； Diabetic nephroathy； Thiamphenicol； Gene expression profiling； Enrichment analysis

糖尿病是一種以高血糖为主要特征的慢性代谢性疾病，引发的各类并发症更加深了糖尿病患者的致死率和致残率[1-4]。其中糖尿病肾病已经成为最常见的糖尿病并发症之一[5-6]。作为严重危害人类健康的疾病，目前对糖尿病肾病治疗仍集中在抗炎、降血糖及血脂和改善血流动力学等方面，缺乏安全有效的治疗药物[7]。近年来，利用基因表达谱技术和关联性图谱（Cmap）数据库筛选和发现药物成为一个新的研究热点[4，8]。因此，为了快速有效地发现新的治疗糖尿病肾病的有效候选药物，本研究从GEO数据库中搜索与糖尿病肾病相关的基因表达谱数据集，筛选出样本数较多的基因芯片：GSE30122；然后利用R软件进行差异基因的分析，获得了与糖尿病肾病相关的差异基因，对差异基因进行分析后，显示甲砜霉素（Thiamphenicol）的负性富集分数最高，表明甲砜霉素具有潜在的改善糖尿病肾病的生物学功能。……