miRNA-451a参与血行播散性肺结核发病机制的初步研究  

A preliminary study on the role of miRNA-451a in the pathogenesis of disseminated tuberculosis

作　　者：赵玲娟 刘浩然 聂文娟[3] 王伟 Zhao Lingjuan;Liu Haoran;Nie Wenjuan;Wang Wei(Department of Emergency,Beijing Chest Hospital,Capital Medical University,Beijing 101149,China;Department of Emergency,Peking Union Medical College Hospital,Beijing 100005,China;Department I of Tuberculosis,Beijing Chest Hospital,Capital Medical University,Beijing 101149,China;National Clinical Laboratory on Tuberculosis,Beijing Chest Hospital,Capital Medical University,Beijing 101149,China)

机构地区：[1]首都医科大学附属北京胸科医院急诊科,北京101149 [2]北京协和医院急诊科,北京100005 [3]首都医科大学附属北京胸科医院结核一科,北京101149 [4]首都医科大学附属北京胸科医院国家结核病临床实验室,北京101149

出　　处：《中国防痨杂志》2025年第5期623-628,共6页Chinese Journal of Antituberculosis

基　　金：临床医学发展专项“扬帆3.0”计划项目(ZLRK202331);首都卫生发展科研专项(2024-1G-2161);国家自然科学基金(82373641)。

摘　　要：目的:初步探讨miRNA-451a参与血行播散性肺结核的发病机制。方法:(1)采用前瞻性研究方法,连续纳入2018年12月至2019年12月就诊于北京胸科医院并接受治疗的继发性肺结核患者56例、血行播散性肺结核(hematogenous disseminated pulmonary tuberculosis,HDPTB)患者24例作为病例组;健康对照组17名均来自本院体检合格者。收集所有研究对象静脉血样本,以及社会人口学信息和临床信息。应用qRT-PCR技术分析外周血中miRNA-451a及其靶基因巨噬细胞迁移抑制因子(macrophage migration inhibitory factor,MIF)的表达水平。(2)培养并分化人急性单核白血病细胞(THP-1),应用细胞转染技术构建miRNA-451a过表达细胞模型,使用MTB标准株H37Rv进行细胞感染,通过qRT-PCR验证基因表达水平,蛋白免疫印迹法验证MIF蛋白表达水平。通过菌落形成单位(colony forming units,CFU)计数检测巨噬细胞对MTB的清除能力。结果:(1)继发性肺结核组、HDPTB组和健康对照组中miRNA-451a的相对表达量分别为24.177(5.150,46.066)、81.742(29.003,117.388)、77.762(54.422,134.633),差异有统计学意义(H=19.040,P<0.001);MIF在三组中的相对表达量分别为0.306(0.168,0.795)、0.189(0.073,0.443)、0.025(0.012,0.085),差异具有统计学意义(H=35.967,P<0.001)。(2)在miRNA-451a过表达的THP-1细胞模型中MIF相对表达水平(0.442±0.019)明显低于阴性对照组(1.477±0.190),差异有统计学意义(t=-9.384,P<0.001);在H37Rv感染细胞24 h后,miRNA-451a过表达巨噬细胞组的总细胞内细菌存活率(42.55%,40/94)明显高于阴性对照组(7.24%,21/290),差异有统计学意义(χ^(2)=66.247,P<0.001)。结论:miRNA-451a通过调节MIF的表达水平进而影响巨噬细胞内结核分枝杆菌的存活情况,参与到HDPTB的发病机制中。Objective:Preliminary exploration of miRNA-451a’s involvement in the pathogenesis of hematogenous disseminated pulmonary tuberculosis.Methods:(1)A prospective research method was adopted,from December 2018 to December 2019,56 patients with secondary pulmonary tuberculosis and 24 patients with hematogenous disseminated pulmonary tuberculosis(HDPTB)treated at Beijing Chest Hospital were consecutively included as the case group,17 healthy controls were selected from those who passed the physical examination in our hospital.Venous blood samples were collected along with sociodemographic and clinical data from all subjects.The expression levels of miRNA-451a and its target gene,macrophage migration inhibitory factor(MIF),in peripheral blood,were analyzed by real-time PCR(qRT-PCR).(2)Human acute monocytic leukemia cells(THP-1)were cultured and differentiated,and the miRNA-451a overexpression cell model was constructed by cell transfection technology,the cells were infected with H37Rv,qRT-PCR verified the gene expression level,and the protein expression level of was verified by Western blotting.The clearance ability of macrophages against H37Rv was determined by colony forming unit(CFU)assay.Results:(1)The relative expression levels of miRNA-451a in the secondary pulmonary tuberculosis patients,HDPTB patients,and healthy controls were 24.177(5.150,46.066),81.742(29.003,117.388),and 77.762(54.422,134.633)respectively,with statistically significant differences(H=19.040,P<0.001);the relative expression levels of MIF were 0.306(0.168,0.795),0.189(0.073,0.443),and 0.025(0.012,0.085)respectively,with statistically significant differences(H=35.967,P<0.001).(2)The relative expression level of MIF in the miRNA-451a overexpressed THP-1 cell model(0.442±0.019)was lower than that in the negative control group(1.477±0.190),with statistically significant differences(t=-9.384,P<0.001);24 hours after H37Rv infection,the total intracellular bacterial survival rate in the miRNA-451a overexpression cell group(42.55%,40/94)was higher t

关 键 词：结核 肺 微RNAS 巨噬细胞游走抑制因子 

分 类 号：R52[医药卫生—内科学]