脓毒症患者血浆循环miRNA表达谱与正常人群差异的初步研究  

Preliminary study on expression profiles of plasma circulating microRNAs in patients with sepsis and healthy control people

作　　者：姚纪友 吕嘉贤 童荔[1] 胡晓光[1] 曹璐[1] 朱艳平[1] 许静红[1] 蔡常洁[1] 

机构地区：[1]中山大学附属第一医院重症二科,广州市510080

出　　处：《实用医学杂志》2017年第24期4024-4028,共5页The Journal of Practical Medicine

基　　金：广州市科学研究专项(编号:201607010186);广州市科技计划项目(编号:201704020153);广东省器官捐献与移植免疫重点实验室建设项目(编号:2013A061401007);广东省器官移植国际合作基地项目(编号:2015B050501002)

摘　　要：目的应用微阵列芯片分析技术筛选脓毒症诊断和预后相关循环miRNAs,通过实时荧光定量PCR验证,并应用生物信息学方法预测脓毒症相关靶基因和发病机制。方法入选脓毒症、脓毒症休克患者及正常人各3例,提取血浆miRNA,分别与第7代人,小鼠和大鼠的miRCURY LNA^(TM)microRNA芯片杂交,扫描信号进行差异表达谱和聚类分析;应用qRT-PCR验证芯片结果。通过miRanda和Targetscan数据库预测miRNAs靶基因及KEGG数据库分析其通路。结果与正常组相比,脓毒症组有57种miRNAs表达上调,脓毒症休克组有11种(fold change≥2.0;P<0.05)。miR-519c-5p被用于qRT-PCR验证,结果表明其在脓毒症中表达明显高于正常组,差异有统计学意义(P<0.05)。通过生物信息学方法我们预测了29个miR-519c-5p相关靶基因,通过KEGG数据库分析其中有7个作用于脓毒症相关通路。最终表明MiR-519c-5p可能正性调节细胞周期调控基因MAP2K4,通过MAPK信号通路导致血管内皮细胞凋亡。结论首次证明血浆miR-519c-5p可能可用于脓毒症诊断并作为可能潜在靶点用于脓毒症治疗。Objective To identify the circulating miRNAs which can be used to evaluate the diagnosis and prognosis of sepsis by microarray and quantitative real-time PCR,and to predict target genes of miR-519c-5p by bioinformatics analysis. Methods Three sepsis patients,3 septic shock patients and 3 normal controls were enrolled in this study. Plasma RNA was extracted,and was used for hybridized by miRCURY LNATMmicroRNA Array.The signals were scanned and used to conduct differential expression profilings and cluster analysis.Further-more,we performed qRT-PCR to confirm the expression of miRNAs chosen from microarray screening. We used the miRanda and Targetscan databases to predict target genes of the concerned miRNAs and used KEGG database to analyze the related pathways. Results Fifty-seven and 11 miRNAs were observed significantly upregulated in sepsis and septic shock patients,respectively（fold change≥2.0;P〈0.05）.qRT-PCR results showed that miR-519c-5p was significantly upregulated in patients with sepsis or patients with septic shock compared with the healthy normal controls（P〈0.05）.Twenty-nine target genes of miR-519c-5p were predicted by the bioinformatics analysis,and 7 potential target genes participate in the sepsis-related pathways.MiR-519c-5p might be a potential positive regulator for the critical cell cycle control gene of MAP2K4,contributing to the vascular endothelial cells apoptosis via MAPK signaling pathway. Conclusions We demonstrated that the plasma level of miR-519c-5p can be used for the diagnosis of sepsis and miR-519c-5p may be a potential therapeutic target for sepsis.

关 键 词：脓毒症 循环miRNA 基因芯片 

分 类 号：R459.7[医药卫生—急诊医学]