基于转录组学数据的抗高尿酸血症肾病药物的预测和活性验证

Prediction and validation of activity of anti-hyperuricemic nephropathy drugs based on transcriptomic data

目的运用转录组学数据从现有药物中筛选可用于抗高尿酸血症肾病(hyperuricemic nephropathy,HN)的小分子药物,为HN的治疗提供新思路。方法从GEO数据获得HN数据集,筛选获得差异表达基因;通过Metascape平台对核心靶点进行KEGG和GO富集分析;采用String数据库构建蛋白质-蛋白质相互作用网络(PPI),Cytoscape计算节点连接度筛选HN发病核心基因;CMAP(connectivity map)数据库中预测抗HN药物,分子对接分析候选药物与核心靶点之间的相互作用;氧嗪酸钾联合腺嘌呤建立HN模型,验证候选药物的肾保护功能。结果从HN转录组数据集GSE190205,共筛选获得557个差异表达基因,其中上调基因374个,下调基因183个,差异基因涉及类风湿性关节炎、脂肪酸降解、TNF信号通路、吞噬小泡等信号通路,ITGAM、TLR2、CD68、CD44和CCL5等基因可能参与HN的发病;通过CMAP分析,筛选到M-3M3FBS、恩扎妥林、白桦脂酸和恩替卡韦等候选药物,进一步动物实验证实恩扎妥林具有良好的肾保护功能,降低HN大鼠的肾脏指数,血清肌酐和尿素氮显著降低(P<0.05或P<0.01),改善肾组织损伤。结论基于转录组学数据的“药物重定位”策略可从现有药物中筛选到有效的抗HN药物,为HN药物的研究提供了新思路。

Objective To screen small molecule drugs that can be used to treat hyperuricemic nephropathy(HN)from existing drugs by using transcriptomic data,providing new ideas for the treatment of HN.Methods HN dataset was obtained from GEO data,and differentially expressed genes were screened.KEGG and GO enrichment analyses were performed on the core target using Metascape platform.The protein-protein interaction network(PPI)was constructed using String database,and Cytoscape calculated node connectivity to screen the core genes of HN pathogenesis.Anti-HN drugs were predicted in CMAP database,and interactions between drug candidates and core targets were analyzed by molecular docking.The HN model was established by potassium oxazinate combined with adenine to validate the renal protective function of candidate drugs.Results A total of 557 differentially expressed genes were screened from HN transcriptome dataset GSE190205,including 374 up-regulated genes and 183 down-regulated genes.The differentially expressed genes were involved in rheumatoid arthritis,fatty acid degradation,TNF signaling pathway,phagosome et al.ITGAM,TLR2,CD68,CD44 and CCL5 genes might be involved in the pathogenesis of HN.By CMAP analysis,candidate drugs such as M-3M3FBS,enzastaurin,betulinate and entecavir were screened.Further animal experiments confirmed that enzastaurin had good renal protection function,significantly decreased the kidney index,serum Cre and serum BUN of HN rats(P<0.05 or P<0.01),and improved the kidney tissue injury.Conclusion The"drug relocation"strategy based on transcriptomic data can screen effective anti-HN drugs from old drugs,which provides a new idea for the study of HN drugs.