婴幼儿呼吸道合胞病毒肺炎外周血单核细胞miR-146a和miR-155的表达变化

The expression changes of miR-146a and miR-155 in peripheral blood mononuclear cells of infants with respiratory syncytial virus pneumonia

目的分析婴幼儿呼吸道合胞病毒(RSV)肺炎外周血单核细胞(PBMCS)中miR-146a和miR-155的表达变化,分析其对疾病的意义。方法选取2017年6月至2018年6月在呼吸科治疗的急性呼吸道RSV阳性患儿110例为RSV组,其中轻症组74例,重症组36例,选择同期来院体检的健康儿童100例为对照组。患儿于入院24 h内,采集鼻咽分泌物,用于RSV抗原检测;抽取研究对象静脉血,分离血清用于酶联免疫吸附法(ELISA)检测血清白介素-1β(IL-1β)、白介素-4(IL-4)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、干扰素-γ(IFN-γ)水平;制备PBMCS,qRT-PCR检测miR-146a和miR-155 mRNA表达水平;Pearson相关性分析方法分析miR-146a和miR-155与炎性因子的相关性;受试者工作特征(ROC)曲线检测PBMCS中miR-146a与miR-155表达水平对RSV肺炎的临床诊断价值。结果qRT-PCR检测显示RSV重症组患儿PBMCS中miR-146a、miR-155水平均显著高于轻症组和对照组儿童,轻症组PBMCS miR-146a、miR-155水平显著高于对照组儿童(P<0.05);细胞因子比较显示,重症组血清TNF-α、IL-1β、IL-4、IL-6水平显著高于轻症组和对照组,轻症组血清TNF-α、IL-1β、IL-4、IL-6水平显著高于对照组,差异均有统计学意义(P<0.05);而重症组血清IFN-γ水平显著低于轻症组,高于对照组,差异均有统计学意义(P<0.05);Pearson法相关性比较显示,miR-146a与TNF-α、IL-1β、IL-4呈正相关(r值分别为0.356、0.412、0.359,P<0.05),与IFN-γ呈负相关(r=0.374,P<0.05);miR-155与TNF-α、IL-1β呈显著正相关(r值分别为0.401、0.512,P<0.05),与IFN-γ呈负相关(r=0.462,P<0.05);ROC分析检测miR-146a、miR-155表达水平对RSV肺炎的诊断价值,结果显示miR-146a的AUC面积为0.881、敏感度为76.0%、特异度为94.0%、截断值为2.601;miR-155的AUC面积为0.910、敏感度为78.0%、特异度为98.0%、截断值为2.048。结论婴幼儿RSV肺炎PBMCS中miR-146a和miR-155的表达水平异常升高,对RSV肺炎疾病发生发展具有重要作用,可为临床早期诊断提供理论基础。

Objective To investigate the expression changes of miR-146a and miR-155 in peripheral blood mononuclear cells(PBMCS)of infants with respiratory syncytial virus(RSV)pneumonia,and to explore their significances in pathogenesis of the diseases.Methods A total of 110 iRSV positive children who were treated in our hospital from June 2017 to June 2018 were enrolled as RSV group,including 74 cases in mild case group,and 36 cases in severe case group,at the same time,the other 100 healthy children who underwent physical examination in our hospital were enrolled as control group.Nasopharyngeal secretions were collected within 24 hours after admission for RSV antigen detection,and the venous blood specimens of these subjects were collected,then the levels of interleukin-1β(IL-1β),interleukin-4(IL-4),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and interferon-γ(IFN-γ)were detected by by enzyme-linked immunosorbent assay(ELISA).Moreover PBMCS was prepared,and the expression levels of miR-146a and miR-155 were detected by qRT-PCR.Pearson correlation analysis was performed to analyze the correlations between miR-146a,miR-155 and inflammatory factors.Inaddition receiver operating characteristic(ROC)curve was used to detect the values of miR-146a and miR-155 levels in PBMCS in the clinical diagnosis of RSV pneumonia.Results The qRT-PCR detection showed that the levels of miR-146a and miR-155 in RSV severe case group were significantly higher than those in mild case group and control group,and the levels of miR-146a and miR-155 in mild case group were significantly higher than those in control group(P<0.05).The cytokine comparison showed that serum levels of TNF-α,IL-1β,IL-4,and IL-6 in severe case group were significantly higher than those in mild case group and control group,and the serum levels of TNF-α,IL-1β,IL-4,and IL-6 in mild case group were significantly higher than those in control group,and the serum levels of IFN-γin severe case group were significantly lower than those in mild case group,but,which were significantly higher than those in control group(P<0.05).Pearson correlation analysis showed that miR-146a was positively correlated with TNF-α,IL-1β,and IL-4,however,which was negatively correlated with IFN-γ.The results of ROC analysis showed that the area of AUC of miR-146a was 0.881,the sensitivity was 76.0%,the specificity was 94.0%,the truncation value was 2.601,and the AUC area of miR-155 was 0.910,the sensitivity was 78.0%,the specificity was 98.0%,the truncation value was 2.048.Conclusion The expression levels of miR-146a and miR-155 in PBMCS of infants with RSV pneumonia are abnormally increased,it plays an important role in the occurrence and development of RSV pneumonia,which can provide theoretical basis for early clinical diagnosis.