多芯片联合药物靶点基因探讨骨关节炎患者滑膜中生物标志物和治疗靶点

Study on Biomarkers and Therapeutic Targets in Synovium of Patients with Osteoarthritis by Multi Chip Combined with Drug Target Gene

目的:探索骨关节炎滑膜组织中的差异基因和相关通路,寻找骨关节炎的关键基因,阐明骨关节炎的发病机制。方法:从GEO数据库中收集4个相关数据集,分别是GSE1919、GSE32317、GSE41038和GSE82107,包括37个骨关节炎滑膜组织样本和16个正常滑膜组织样本,鉴定出差异表达基因(DEGs)。对这些DEGs进行GO功能富集分析和KEGG通路富集分析,使用Cytoscape软件构建DEGs的蛋白质相互作用(PPI)网络,筛选Degree前10位的基因。通过Drugbank数据库筛选获准的用于治疗骨关节炎的药物,Drug Gene Interaction数据库挖掘这些药物的靶基因。利用微生信在线作图工具将药物靶基因和PPI Degree前10位的基因取交集,获得药物靶基因与PPI Degree前10位基因相同的基因靶点。最后,再利用定量逆转录聚合酶链反应(qRT-PCR)对临床样本检测评估关键基因的基因表达量。结果:总共鉴定出68个DEGs,包括29个上调的DEGs(MRC2、CDH11、HK3等)和39个下调的DEGs(ADH1B、APOD、ADIPOQ等)。其功能主要富集在皮质类固醇的反应、血液循环、氧化还原酶活性和冷诱导产热的正向调节过程,以及IL^(-1)7信号通路、PPAR信号通路、肿瘤坏死因子信号通路和糖酵解。PPI Degree前10位的基因分别是LEP、MMP-9、FOS、CXCL10、MMP-1、TNFSF11、ADIPOQ、FABP4、LPL和MMP-3。通过Drugbank数据库和Drug Gene Interaction数据库共挖掘出30种获得批准的治疗骨关节炎药物及对应的284个靶基因。利用微生信在线作图工具,获得治疗骨关节炎药物靶基因与PPI Degree前10位的基因交集为MMP-9、LPL和MMP-1。定量逆转录聚合酶链反应结果显示,MMP-9在骨关节炎患者滑膜中呈高表达,且差异有统计学意义(P<0.01);LPL在骨关节炎患者滑膜中呈低表达,且差异有统计学意义(P<0.05)。结论:LPL和MMP-9是用于治疗骨关节炎的有效分子靶标。

Objective:To explore the differential genes and related pathways in the synovium of osteoarthritis,find the key genes of osteoarthritis,and clarify the pathogenesis of osteoarthritis.Methods:Four related data sets were collected from GEO database,namely GSE1919,GSE32317,GSE41038 and GSE82107,including 37 synovial tissue samples of osteoarthritis and 16 normal synovial tissue samples,and differential expression genes(DEGs)were identified.GO function enrichment analysis and KEGG pathway enrichment analysis were carried out for these DEGs,and protein interaction(PPI)network of DEGs was constructed using software Cytoscape to screen the top 10 genes of Degree.The Drugbank database was used to screen approved drugs for the treatment of osteoarthritis,and the Drug Gene Interaction database was used to mine the target genes of these drugs.The drug target gene and the top 10 genes of PPI Degree were intersected by using the online mapping tool of Bioinformatics to obtain the same gene target of the drug target gene and the top 10 genes of PPI Degree.Finally,quantitative reverse transcription polymerase chain reaction(q RT-PCR)was used to detect and evaluate the gene expression of key genes in clinical samples.Results:A total of 68 DEGs were identified,including 29 up regulated DEGs(MRC2,CDH11 and HK3)and 39 down regulated DEGs(ADH1B,APOD and ADIPOQ).Their functions are mainly concentrated in the positive regulation process of corticosteroid response,blood circulation,oxidoreductase activity and cold induced thermogenesis,as well as IL^(-1)7 signal pathway,PPAR signal pathway,tumor necrosis factor signal pathway and glycolysis.The first 10 genes of PPI Degree are respectively LEP,MMP-9,FOS,CXCL10,MMP-1,TNFSF11,ADIPOQ,FABP4,LPL and MMP-3.Through the Drugbank database and the Drug Gene Interaction database,a total of 30 approved drugs for the treatment of osteoarthritis and their corresponding 284 target genes were mined.Using the online mapping tool of Bioinformatics,the intersection(MMP-9,LPL and MMP-1)of the drug target gene for osteoarthritis and the top 10 genes of PPI Degree were obtained.Quantitative reverse transcription polymerase chain reaction showed that MMP-9 was highly expressed in synovium of patients with osteoarthritis,and the difference was statistically significant(P<0.01);LPL was expressed low in synovium of patients with osteoarthritis,and the difference was statistically significant(P<0.05).Conclusion:LPL and MMP-9 are effective molecular targets for the treatment of osteoarthritis.