生物信息学分析及实验验证探索抗中性粒细胞胞质抗体相关性血管炎炎症应答候选基因及分子机制  

Identification of inflammatory response genes in ANCA-associated vasculitis by bioinformatics analysis and experimental validation

作　　者：张冬梅 张妍楠 秦建华 欧三桃 吴蔚桦 Zhang Dongmei;Zhang Yannan;Qin Jianhua;Ou Santao;Wu Weihua(Dept of Nephrology,The Affiliated Hospital of Southwest Medical University,Luzhou 646000;Sichuan Clinical Research Center for Nephropathy,Luzhou 646000)

机构地区：[1]西南医科大学附属医院肾病内科,泸州646000 [2]四川省肾脏疾病临床研究中心,泸州646000

出　　处：《安徽医科大学学报》2024年第4期581-589,共9页Acta Universitatis Medicinalis Anhui

基　　金：四川省卫生健康委员会医学科技项目(编号:21PJ099)。

摘　　要：目的通过生物信息学方法及实验验证探索抗中性粒细胞胞质抗体(ANCA)相关性血管炎炎症应答候选基因及潜在的分子机制,为治疗ANCA相关性血管炎潜在炎症靶标提供科学的理论依据。方法从GEO数据库检索获得GSE108109芯片数据,利用R语言相关程序包处理、分析并筛选出差异基因。利用DAVID在线网站进行京都基因与基因组百科全书(KEGG)和基因本体(GO)富集分析,并通过STRING网站构建炎症候选基因编码蛋白的相互作用网络。进一步通过miRWalk和DIANA-LncBase数据库预测并构建内源性竞争性RNA(ceRNA)调控网络,并从网络中筛选出关键基因绘制ROC曲线。纳入西南医科大学附属医院确诊并经肾穿刺活检证实的ANCA相关性血管炎患者肾组织标本进行验证,以非ANCA相关性血管炎患者肾组织标本(IgA肾病、微小病变型肾病)为对照组。对收集到的肾组织标本进行免疫组化染色,并通过免疫组化染色半定量分析计算平均光密度,进一步验证生信分析筛选出的关键基因的表达情况,同时将关键基因的平均光密度值与炎症指标进行Pearson线性相关性分析。结果共筛选出差异表达基因846个,其中444个基因表达明显上调,402个基因表达明显下调。通过KEGG和GO富集分析获得了与炎症调控相关的重要差异表达基因,其中CSF1R和TNFRSF1B为首次在ANCA相关性血管炎中报道的差异基因。同时构建了包括KCNQ1OT1-hsa-miR-125a-5p-TNFRSF1B在内的多条内源性竞争RNA(ceRNA)调控轴。收集到ANCA相关性血管炎标本15例,IgA肾病标本6例,微小病变型肾标本3例。肾穿组织标本的免疫组化结果提示CSF1R、TNFRSF1B在ANCA相关性血管炎肾组织表达较对照组均有升高,对ANCA组患者临床数据做Pearson相关性分析,得出CSF1R的表达量与中性粒细胞计数含量呈正相关(r=0.587),TNFRSF1B的表达量和血清C反应蛋白含量呈正相关(r=0.646)。结论通过生物信息学的�Objective To explore the candidate genes and potential molecular mechanisms of anti-neutrophil cytoplasmic antibodies(ANCA)-associated vasculitis by bioinformatics and experimental validation,and to provide a scientific theoretical basis for the treatment of potential inflammatory targets for ANCA-associated vasculitis.Methods The GSE108109 chip data was retrieved from the Gene Expression Omnibus(GEO)database,and the differential genes were processed,analyzed and screened using the R language related program package.Kyoto encyclopedia of genes and genomes(KEGG)and gene ontology(GO)enrichment analysis was carried out using DAVID online network cable,and the interaction network of the protein encoded by the selected genes of inflammatory syndrome was constructed through STRING website.Further endogenous competitive RNA(ceRNA)regulatory network was predicted and constructed through miRWalk and DIANA-LncBase databases,and key genes were screened from the network to draw ROC curve.The renal biopsy samples of patients with ANCA-associated vasculitis confirmed by our hospital were collected as the experimental group,and the renal biopsy samples of IgA nephropathy and micro-adaptive nephropathy were collected as the control group.Immunohistochemical staining was performed on the collected renal biopsy samples,and the average optical density was calculated by semi-quantitative analysis of immunohistochemical staining to further verify the expression of the key genes screened by the bioinformatics analysis.Pearson linear correlation analysis was performed between the average optical density results and the clinical inflammatory data of patients.Results 846 differential genes were screened,of which 444 genes were significantly up-regulated and 402 genes were significantly down-regulated.Through KEGG and GO analysis,important differentially expressed genes related to inflammation regulation were obtained.Among them,CSF1R and TNFRSF1B,two differentially expressed genes never reported in ANCA-associated vasculitis,attracted ou

关 键 词：ANCA相关性血管炎 生物信息学分析 CSF1R TNFRSF1B 炎症 内源性竞争RNA 

分 类 号：R593.2[医药卫生—内科学]