川崎病患儿尿液外泌体中差异表达miRNA的筛选意义

Significance of screening differentially expressed miRNA in urine exosomes of children with Kawasaki disease

目的研究川崎病(KD)患儿尿液外泌体中差异表达的微小RNA(miRNA),探讨其作为KD诊断新型分子标志物的可能性。方法选取2022年1月至12月在西安市第三医院儿科住院的15例KD患儿作为试验组,另选取同期在儿童保健科体检的15例正常儿童作为对照组。采用试剂法从两组患儿中分别提取3组中段晨尿中的外泌体;采用高通量测序分析检测两组尿液外泌体中的差异表达miRNA,并对其进行GO、KEGG通路分析。比较两组的miRNA差异表达情况。结果共筛选出29种上调差异表达miRNA。GO富集分析发现,差异miRNA参与的生物学过程(BP)包括信号转导、DNA转录调控、RNA聚合酶Ⅱ转录调控、细胞分化、磷酸化过程等;细胞组分(CC)包括细胞膜、细胞质、细胞核等;分子功能(MF)包括蛋白结合、金属离子结合、DNA结合等。KEGG通路分析发现,差异miRNA明显富集的信号通路是细胞因子-细胞因子受体相互作用、配体-受体相互作用、物质转运和分解代谢、细胞内信号传导、肿瘤、病毒感染等。结论尿液外泌体中差异表达的miRNA可作为诊断KD的新型分子标志物。

Objective To study the differential expression of microRNA(miRNA)in urine exosomes of children with Kawasaki disease(KD),and to explore its possibility as a new molecular marker for KD diagnosis.Methods A total of 15 children with KD who were hospitalized in the pediatrics department of the Xi'an No.3 Hospital from January to December 2022 were selected as experimental group,and 15 normal children who underwent physical examination in the child health care department during the same period were selected as control group.Exosomes were extracted from 3 mid-morning urine samples of the two groups by reagent method;high-throughput sequencing analysis was used to detect differentially expressed miRNA in urine exosomes of the two groups,and GO and KEGG pathway analysis were performed.The differential expression of miRNA between the two groups was compared.Results A total of 29 up-regulated differentially expressed miRNA were screened.GO enrichment analysis showed that the biological process(BP)involved in differential miRNA included signal transduction,DNA transcription regulation,RNA polymerase Ⅱ transcription regulation,cell differentiation and phosphorylation process,etc.;cellular component(CC)included cell membrane,cytoplasm and nucleus,etc.;molecular function(MF)included protein binding,metal ion binding and DNA binding,etc.KEGG pathway analysis showed that the signaling pathways significantly enriched by differential miRNA were cytokine-cytokine receptor interaction,ligand-receptor interaction,material transport and catabolism,intracellular signal transduction,tumor and virus infection,etc.Conclusion The differentially expressed miRNA in urine exosomes can be used as a new molecular marker for the diagnosis of KD.