CCL20通过IL-17信号通路调控糖尿病肾病炎症进展

CCL20 regulates inflammatory progression in diabetic nephropathy through IL-17 signaling pathway

目的基于加权基因共表达网络分析(WGCNA)识别糖尿病肾病(DN)外周血中的关键基因,探讨DN的发病机制,为其发生、发展提供新的见解。方法通过基因表达总库(GEO)下载数据集GSE142153进行差异表达基因(DEGs)筛选,应用京都基因与基因组百科全书(KEGG)通路富集分析DEGs参与的分子生物学过程。通过构建WGCNA识别关键模块和中枢(hub)基因,应用验证集GSE65561分析受试者操作特征曲线从而验证hub基因潜在的临床诊断价值;随后采用实时荧光定量PCR(RT-qPCR)检测hub基因在DN患者外周血中的相对表达量。结果数据集GSE142153包含23例DN患者、10例健康对照者。根据筛选标准,DN组有181个DEGs,其中106个基因表达上调和75个基因表达下调。KEGG富集分析聚焦于IL-17、核因子-κB、酪氨酸蛋白激酶/信号转导及转录激活因子等炎症通路激活。WGCNA识别了16个模块,其中,turquoise模块被认为对DN最具临床意义,选择turquoise模块中的基因与DEGs、IL-17信号通路基因相交得C-C趋化因子配体20(CCL20)、IL-1β、C-X-C趋化因子配体1、基质金属蛋白酶-9、FOS样抗原1基因。临床DN患者外周血RT-qPCR验证,结果与生物信息学分析一致。结论CCL20等hub基因可能通过调控IL-17信号通路促进DN的炎症发展,该类基因有助于确定DN的潜在生物标志物和治疗靶点。

Objective To identify key genes in the peripheral blood of diabetic nephropathy(DN)patients based on weighted gene co-expression network analysis(WGCNA),aiming to explore the pathogenesis of DN and provide new insights into the incidence and development of DN.Methods In this study,the dataset GSE142153 was downloaded from the Gene Expression Omnibus(GEO)database to screen the differentially expressed genes(DEGs).Then,Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis was employed to investigate the molecular biological processes of these DEGs.Key modules and hub genes were identified by constructing WGCNA.The validation dataset GSE65561 was applied to analyze the receiver operating characteristic(ROC)curve to validate the potential clinical diagnostic value of hub genes.Subsequently,RT-qPCR was adopted to detect the relative expression levels of hub genes in the peripheral blood of DN patients.Results There were 23 patients with DN and 10 healthy controls in the dataset GSE142153.According to the screening criteria,there were 181 DEGs in the DN group,of which 106 DEGs were up-regulated and 75 were down-regulated.KEGG enrichment analysis mainly showed the activation of inflammatory pathways,such as interleukin(IL)-17,nuclear factor-κB(NF-κB),and JAK-STAT signaling pathways.Sixteen modules were identified by WGCNA and the turquoise module was considered to have the most clinical significance for DN.The genes in the turquoise module were selected to intersect with DEGs and IL-17 signaling pathway-related genes to obtain 5 hub genes including Chemokine(C-C motif)ligand 20(CCL20),IL-1B,CXCL1,matrix metalloproteinase-9(MMP-9)and Fos-like antigen 1(FOSL1).The results of RT-qPCR of the peripheral blood of DN patients were consistent with the bioinformatic analysis.Conclusion The hub genes,such as CCL20,may play a key role in promoting the inflammatory progression of DN through regulating the IL-17 signaling pathway,which contribute to identifying potential biomarkers and therapeutic targets for DN.