外周血单个核细胞中微小RNA200c-3p、核因子κB mRNA和膜联蛋白-5 mRNA对儿童哮喘诊断价值的比较研究

Comparative study on the diagnostic value of miRNA-200c-3p, NF-κB mRNA and annexin-5 mRNA in peripheral blood mononuclear cells for childhood asthma

摘要

目的 比较外周血单个核细胞(PBMCs)中微小RNA200c-3p(miRNA-200c-3p)、核因子κB(NF-κB)mRNA和膜联蛋白-5(ANX-5)mRNA对儿童哮喘的诊断价值。方法 纳入2022年12月至2023年12月间在唐山市妇幼保健院儿科就诊的95例哮喘儿童作为哮喘组,并分成轻度组(30例),中度组(36例),重度组(29例),同期来该院体检健康儿童40例为对照组。使用实时荧光定量逆转录聚合酶链反应(qRT-PCR)法检测单个核细胞中miRNA-200c-3p、NF-κB mRNA和ANX-5 mRNA的表达量。测量肺功能、1s用力呼气量(FEV₁)、用力肺活量(FVC)、最大呼气流量(PEF)及FEV₁/FVC值。采用Spearman分析miRNA-200c-3p、NF-κB mRNA和ANX-5 mRNA与肺功能的相关性。使用受试者工作特征(ROC)曲线比较miRNA-200c-3p、NF-κB mRNA和ANX-5 mRNA对哮喘的诊断价值。结果 miRNA-200c-3p和NF-κB mRNA表达水平在重度组、中度组、轻度组及对照组间差异显著(F=12.58和14.81,P<0.01),并依次降低。ANX-5 mRNA表达水平在四组间差异显著(F=16.34,P<0.01),并依次升高。哮喘患儿单个核细胞中miRNA-200c-3p和NF-κB mRNA,与FEV₁、PEF、FEV₁/FVC均呈负相关(r介于-0.80~-0.48,P<0.05),ANX-5 mRNA与FEV₁、PEF、FEV₁/FVC呈正相关(r介于0.48~0.83,P<0.001)。ROC曲线显示,外周血PBMCs中miRNA-200c-3p诊断儿童哮喘的AUC为0.858,敏感度为0.760,特异度为0.893,截断值为2.24,高于NF-κB mRNA和ANX-5 mRNA单独诊断儿童哮喘。结论 单个核细胞中miRNA-200c-3p对儿童哮喘的诊断效能优于NF-κB mRNA及ANX-5 mRNA单独诊断,是儿童哮喘诊断的敏感分子标志物。

Abstract

Objective To compare the diagnostic value of miRNA-200c-3p, nuclear factor κB (NF-κB) mRNA and annexin-5 (ANX-5) mRNA in peripheral blood mononuclear cells (PBMCs) for childhood asthma. Methods A total of 95 children with asthma who visited the Department of Pediatrics at Tangshan Maternal and Child Health Hospital between December 2022 and December 2023 were included as the asthma group. This group was further divided into a mild group (30 cases), a moderate group (36 cases), and a severe group (29 cases). Concurrently, 40 healthy children who underwent physical examinations at the hospital were selected as the control group. The expression levels of miRNA-200c-3p, NF-κB mRNA, and ANX-5 mRNA in PBMCs were measured using real-time fluorescence quantitative reverse transcription polymerase chain reaction (qRT-PCR). Pulmonary function was assessed by measuring forced expiratory volume in 1 second (FEV ₁), forced vital capacity (FVC), peak expiratory flow (PEF), and the FEV₁/FVC ratio. Spearman correlation analysis was used to evaluate the relationships between miRNA-200c-3p, NF-κB mRNA, ANX-5 mRNA, and pulmonary function. The diagnostic value of miRNA-200c-3p, NF-κB mRNA, and ANX-5 mRNA for asthma was compared using receiver operating characteristic (ROC) curve analysis. Results The expression levels of miRNA-200c-3p and NF-κB mRNA differed significantly among the severe, moderate, mild, and control groups (F=12.58 and 14.81, respectively, P<0.01), decreasing sequentially across the groups. In contrast, ANX-5 mRNA expression levels showed significant differences among the four groups (F=16.34, P<0.01), increasing sequentially. In children with asthma, miRNA-200c-3p and NF-κB mRNA were negatively correlated with FEV₁, PEF, and FEV₁/FVC (r ranging from -0.80 to -0.48, P<0.05), while ANX-5 mRNA was positively correlated with FEV₁, PEF, and FEV₁/FVC (r ranging from 0.48 to 0.83, P<0.001). The ROC curve analysis showed that miRNA-200c-3p in PBMCs had the highest diagnostic value for childhood asthma, with an area under the curve (AUC) of 0.858, sensitivity of 0.760, specificity of 0.893, and a cutoff value of 2.24, outperforming NF-κB mRNA and ANX-5 mRNA as standalone diagnostic markers. Conclusion miRNA-200c-3p in mononuclear cells demonstrates superior diagnostic performance for childhood asthma compared to NF-κB mRNA and ANX-5 mRNA alone, making it a sensitive molecular biomarker for the diagnosis of childhood asthma.

关键词

儿童 / 哮喘 / 单个核细胞 / 微小RNA-200c-3p / 膜联蛋白-5 / NF-κB / 诊断价值

Key words

child / asthma / peripheral blood mononuclear cell / miRNA-200c-3p / annexin-5 / NF-κB / diagnostic value

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基金资助

河北省医学科学研究课题(20221758)