利用肾小球全基因组表达数据探索糖尿病肾病治疗药物的研究

发布时间：2018-10-25 16:03

【摘要】：目的:利用糖尿病肾病(diabetic nephropathy,DN)患者微分离的肾小球全基因组表达数据,借助CMAP(Connectivity Map)数据库和生物信息学方法,探寻具有DN治疗作用的潜在药物。方法:选取中国汉族DN患者29例和正常对照6例,获取肾组织后,激光微分离肾小球,提取RNA后,利用Affymetrix U133 Plus 2.0全基因组表达谱芯片检测获得基因表达数据,获得不同分期DN患者之间的差异表达基因后,进一步通过2种不同的生物信息学方法寻找DN的潜在治疗药物;并阐明DN治疗药物可能的分子作用机制。结果:1.与正常对照相比,早期和晚期DN患者肾小球的差异表达基因分别为100和1602个(FDR0.05,变化倍数1.5);DN晚期和早期相比,肾小球有1194个基因出现差异表达(FDR0.05,变化倍数1.5)。对DN晚期和早期相比的差异表达基因进行功能分析,基因本体论(Gene ontology,GO)分析显示,差异表达基因主要涉及细胞外刺激、组织生长发育、免疫和细胞信号传导等,进一步的通路分析显示细胞外基质受体相互作用、细胞与细胞外基质的连接、补体系统、细胞因子受体相互作用和PI3K-Akt信号通路等出现明显变化;2.利用DN晚期和早期相比的差异表达基因和CMAP数据库,筛选出小白菊内酯(parthenolide)、荜茇酰胺(piperlongumine)、15d-PGJ2(15-脱氧前列腺素 J2)和 LY-294002(PIBK抑制剂)等候选药物具有逆转晚期DN患者基因变化的作用,提示上述药物可能具有DN治疗的潜能,进一步的文献分析表明这些药物对DN均有一定的治疗作用,可能作为NF-kB抑制剂、组蛋白去乙酰化酶抑制剂、PI3K信号通路抑制剂或者PPARγ激动剂等发挥肾脏保护作用。结论:利用DN患者肾小球全基因组表达谱和CMAP数据库能够快速筛选出具有DN治疗潜能的药物;文献分析也验证了该方法的可靠性;相关药物可以进行下一步的动物和临床试验以证实其治疗的安全性和有效性。
[Abstract]:Aim: to explore the potential drugs for the treatment of diabetic nephropathy (diabetic nephropathy,DN) by using microisolated glomerular genomic expression data, CMAP (Connectivity Map) database and bioinformatics. Methods: 29 patients with DN in Chinese Han nationality and 6 normal controls were selected. After renal tissue was obtained, glomeruli were isolated by laser microseparation, RNA was extracted, and gene expression data were obtained by Affymetrix U133 Plus 2.0 whole genome expression microarray. After obtaining the differentially expressed genes among DN patients at different stages, we further explored the potential therapeutic drugs for DN by two different bioinformatics methods, and elucidated the possible molecular mechanism of DN treatment drugs. The result is 1: 1. Compared with normal controls, the differential expression genes in glomeruli of early and late DN patients were 100,1602 (FDR0.05, variation multiple 1.5); DN late and early stage respectively), and there were 1194 differentially expressed genes (FDR0.05, change multiple 1.5) in glomeruli of early and late DN patients. Functional analysis of differentially expressed genes in late and early stages of DN showed that differentially expressed genes were mainly involved in extracellular stimulation, tissue growth and development, immunity and cell signal transduction. Further pathway analysis showed that there were significant changes in extracellular matrix receptor interaction, cell / extracellular matrix connection, complement system, cytokine receptor interaction and PI3K-Akt signaling pathway. 2. By using the differentially expressed genes and CMAP database of late and early stage of DN, the candidate drugs such as (parthenolide), lonymine (piperlongumine), 15d-PGJ2 (15-deoxyprostaglandin J _ 2) and LY-294002 (PIBK inhibitor) could reverse the gene changes in patients with advanced DN. It is suggested that these drugs may have the potential of DN therapy. Further literature analysis shows that these drugs have some therapeutic effects on DN and may be used as NF-kB inhibitors. Histone deacetylase inhibitors, PI3K signaling pathway inhibitors or PPAR 纬 agonists play a role in renal protection. Conclusion: the genome-wide expression profile and CMAP database of DN patients can be used to quickly screen the drugs with DN therapeutic potential, and the reliability of the method is verified by literature analysis. Related drugs can be tested in future animal and clinical trials to demonstrate the safety and efficacy of the treatment.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R587.2;R692.9

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