Department of Tuberculosis Treatment and Prevention, Shaanxi Provincial Institute for Tuberculosis Control and Prevention, Xi'an, Shaanxi 710048, P.R. China.
Department of Orthopedics, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
Mol Med Rep. 2021 Apr;23(4). doi: 10.3892/mmr.2021.11888. Epub 2021 Feb 4.
Pulmonary tuberculosis (TB) is a chronic respiratory infectious disease. Certain microRNAs (miRNAs or miRs) are reported to be involved in regulating TB progression. The present study aimed to evaluate the diagnostic potential of miR‑125b in pulmonary TB. The expression levels of miR‑125b and Raf1 proto‑oncogene serine/threonine protein kinase (RAF1) were analyzed via reverse transcription‑quantitative (RT‑q)PCR in patients with TB. The correlation between miR‑125b and the clinical indicators was investigated, and a receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of miR‑125b. The relationship between miR‑125b and RAF1 was examined using the dual luciferase reporter gene assay. IL‑6, TNF‑α, NF‑κB and IFN‑γ levels were detected using ELISA kits, and then the correlation between miR‑125b expression and the levels of IL‑6, TNF‑α, NF‑κB, IFN‑γ and RAF1 in peripheral blood mononuclear cells (PBMCs) was analyzed. Moreover, RAF1 mRNA and protein expression levels were detected via RT‑qPCR and western blotting. The results demonstrated that miR‑125b expression was decreased in patients with TB, while RAF1 expression was increased. Furthermore, miR‑125b expression was associated with sputum acid‑fast bacillus smear. The area under the ROC curve of miR‑125b was 0.9413, and the sensitivity and specificity of miR‑125b expression for TB were 90 and 92.5%, respectively. IL‑6, TNF‑α, NF‑κB and IFN‑γ levels were negatively correlated with miR‑125b expression, and were inhibited by miR‑125b in PBMCs. Moreover, miR‑125b targeted RAF1 to negatively regulate its expression levels. RAF1 reversed the role of miR‑125b in attenuating IL‑6, TNF‑α, NF‑κB and IFN‑γ levels in PBMCs. The present study demonstrated that the levels of IL‑6, TNF‑α, NF‑κB and IFN‑γ were negatively correlated with miR‑125b expression in PBMCs. Thus, it was suggested that miR‑125b served important roles in the occurrence and development of TB by decreasing the levels of IL‑6, TNF‑α, NF‑κB and IFN‑γ by inhibiting RAF1.
肺结核（TB）是一种慢性呼吸道传染病。有研究报道某些 microRNAs（miRNAs 或 miRs）参与调控 TB 的进展。本研究旨在评估 miR-125b 在肺结核中的诊断潜力。通过逆转录定量（RT-q）PCR 分析 TB 患者中 miR-125b 和 Raf1 原癌基因丝氨酸/苏氨酸蛋白激酶（RAF1）的表达水平。研究了 miR-125b 与临床指标的相关性，并使用受试者工作特征（ROC）曲线评估 miR-125b 的诊断价值。采用双荧光素酶报告基因检测 miR-125b 与 RAF1 之间的关系。使用 ELISA 试剂盒检测白细胞介素 6（IL-6）、肿瘤坏死因子-α（TNF-α）、核因子-κB（NF-κB）和干扰素-γ（IFN-γ）水平，然后分析 miR-125b 表达与外周血单个核细胞（PBMC）中 IL-6、TNF-α、NF-κB、IFN-γ和 RAF1 水平之间的相关性。此外，通过 RT-qPCR 和 Western blot 检测 RAF1 mRNA 和蛋白表达水平。结果表明，TB 患者 miR-125b 表达降低，而 RAF1 表达升高。此外，miR-125b 表达与痰抗酸杆菌涂片有关。miR-125b 的 ROC 曲线下面积为 0.9413，miR-125b 表达对 TB 的灵敏度和特异性分别为 90%和 92.5%。IL-6、TNF-α、NF-κB 和 IFN-γ 水平与 miR-125b 表达呈负相关，并可被 miR-125b 在 PBMCs 中抑制。此外，miR-125b 靶向 RAF1 以负向调控其表达水平。RAF1 逆转了 miR-125b 在降低 PBMCs 中 IL-6、TNF-α、NF-κB 和 IFN-γ 水平中的作用。本研究表明，IL-6、TNF-α、NF-κB 和 IFN-γ 水平与 PBMCs 中 miR-125b 表达呈负相关。因此，miR-125b 通过抑制 RAF1 降低 IL-6、TNF-α、NF-κB 和 IFN-γ 水平，从而在 TB 的发生和发展中发挥重要作用。
