miRNA-451a参与血行播散性肺结核发病机制的初步研究

doi:10.19982/j.issn.1000-6621.20240524

中国防痨杂志 ›› 2025, Vol. 47 ›› Issue (5): 623-628.doi: 10.19982/j.issn.1000-6621.20240524

miRNA-451a参与血行播散性肺结核发病机制的初步研究

赵玲娟¹, 刘浩然², 聂文娟³(), 王伟⁴()

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

收稿日期:2024-11-26 出版日期:2025-05-10 发布日期:2025-04-29
通信作者: 王伟,Email:wangwei010@aliyun.com;聂文娟,Email:94642975@qq.com
基金资助:
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)

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

Zhao Lingjuan¹, Liu Haoran², Nie Wenjuan³(), Wang Wei⁴()

¹Department of Emergency, Beijing Chest Hospital, Capital Medical University, Beijing 101149, China
²Department of Emergency, Peking Union Medical College Hospital, Beijing 100005, China
³Department I of Tuberculosis, Beijing Chest Hospital, Capital Medical University, Beijing 101149, China
⁴National Clinical Laboratory on Tuberculosis, Beijing Chest Hospital, Capital Medical University, Beijing 101149, China

Received:2024-11-26 Online:2025-05-10 Published:2025-04-29
Contact: Wang Wei, Email:wangwei010@aliyun.com; Nie Wenjuan, Email: 94642975@qq.com
Supported by:
Clinical Medicine Development Special Project “Sailing 3.0” Plan Project(ZLRK202331);Capital’s Funds for Health Improvement and Research(2024-1G-2161);National Natural Science Foundation of China(82373641)

摘要/Abstract

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24h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(χ²=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。

关键词: 结核,肺, 微RNAs, 巨噬细胞游走抑制因子

Abstract:

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 than that in the negative control group (7.24%, 21/290) with statistically significant differences (χ²=66.247, P<0.001). Conclusion: miRNA-451a is involved in the pathogenesis of HDPTB by regulating the expression of MIF and then affecting the survival of bacteria in macrophages.

Key words: Tuberculosis, pulmonary, MicroRNAs, Macrophage migration-inhibitory factors

中图分类号:

赵玲娟, 刘浩然, 聂文娟, 王伟. miRNA-451a参与血行播散性肺结核发病机制的初步研究[J]. 中国防痨杂志, 2025, 47(5): 623-628. doi: 10.19982/j.issn.1000-6621.20240524

Zhao Lingjuan, Liu Haoran, Nie Wenjuan, Wang Wei. A preliminary study on the role of miRNA-451a in the pathogenesis of disseminated tuberculosis[J]. Chinese Journal of Antituberculosis, 2025, 47(5): 623-628. doi: 10.19982/j.issn.1000-6621.20240524

图/表 1

参考文献 25

[1]	Suárez I, Maria Fünger S, Jung N, et al. Severe disseminated tuberculosis in HIV-negative refugees. Lancet Infect Dis, 2019, 19(10):e352-e359. doi:10.1016/S1473-3099(19)30162-8.
[2]	Meira L, Chaves C, Araújo D, et al. Predictors and outcomes of disseminated tuberculosis in an intermediate burden setting. Pulmonology, 2019, 25(6): 320-327. doi:10.1016/j.pulmoe.2018.11.001. pmid: 30819659
[3]	Sharma S K, Mohan A, Sharma A. Miliary tuberculosis: A new look at an old foe. J Clin Tuberc Other Mycobact Dis, 2016, 3: 13-27. doi:10.1016/j.jctube.2016.03.003. pmid: 31723681
[4]	Sharma SK, Mohan A, Sharma A, et al. Miliary tuberculosis: new insights into an old disease. Lancet Infect Dis, 2005, 5(7): 415-430. doi:10.1016/S1473-3099(05)70163-8. pmid: 15978528
[5]	Wei X, Xie M, Wu S, et al. The clinical features and prognostic factors of miliary tuberculosis in a high tuberculosis burden area. Ann Med, 2024, 56(1): 2356647. doi:10.1080/07853890.2024.2356647.
[6]	Wang JY, Hsueh PR, Wang SK, et al. Disseminated tuberculosis: a 10-year experience in a medical center. Medicine (Baltimore), 2007, 86(1): 39-46. doi:10.1097/MD.0b013e318030b605.
[7]	Moule MG, Cirillo JD. Mycobacterium tuberculosis Dissemination Plays a Critical Role in Pathogenesis. Front Cell Infect Microbiol, 2020, 10: 65. doi:10.3389/fcimb.2020.00065.
[8]	Godlewski J, Nowicki MO, Bronisz A, et al. MicroRNA-451 regulates LKB1/AMPK signaling and allows adaptation to metabolic stress in glioma cells. Mol Cell, 2010, 37(5): 620-632. doi:10.1016/j.molcel.2010.02.018. pmid: 20227367
[9]	Lin Z, Xie X, Gu M, et al. microRNA-144/451 decreases dendritic cell bioactivity via targeting interferon-regulatory factor 5 to limit DSS-induced colitis. Front Immunol, 2022, 13: 928593. doi:10.3389/fimmu.2022.928593.
[10]	Gilfillan M, Das P, Shah D, et al. Inhibition of microRNA-451 is associated with increased expression of Macrophage Migration Inhibitory Factor and mitgation of the cardio-pulmonary phenotype in a murine model of Bronchopulmonary Dysplasia. Respir Res, 2020, 21(1): 92. doi:10.1186/s12931-020-01353-9.
[11]	Das R, Koo MS, Kim BH, et al. Macrophage migration inhibi-tory factor (MIF) is a critical mediator of the innate immune response to Mycobacterium tuberculosis. Proc Natl Acad Sci U S A, 2013, 110(32): E2997-3006. doi:10.1073/pnas.1301128110.
[12]	中华人民共和国国家卫生和计划生育委员会. WS 288—2017 肺结核诊断. 结核与肺部疾病杂志, 2024, 5(4): 376-378. doi:10.19983/j.issn.2096-8493.2024022.
[13]	Roger T, David J, Glauser MP, et al. MIF regulates innate immune responses through modulation of Toll-like receptor 4. Nature, 2001, 414(6866): 920-924. doi:10.1038/414920a.
[14]	Roger T, Froidevaux C, Martin C, et al. Macrophage migration inhibitory factor (MIF) regulates host responses to endotoxin through modulation of toll-like receptor 4 (TLR4). J Endotoxin Res, 2003, 9(2): 119-123. doi:10.1179/09680510 3125001513. pmid: 12803886
[15]	Mitchell RA, Liao H, Chesney J, et al. Macrophage migration inhibitory factor (MIF) sustains macrophage proinflammatory function by inhibiting p53: Regulatory role in the innate immune response. Proc Natl Acad Sci U S A, 2002, 99(1): 345-350. doi:10.1073/pnas.012511599.
[16]	Calandra T, Roger T. Macrophage migration inhibitory factor: A regulator of innate immunity. Nat Rev Immunol, 2003, 3(10): 791-800. doi:10.1038/nri1200. pmid: 14502271
[17]	Donnelly SC, Haslett C, Reid PT, et al. Regulatory role for macrophage migration inhibitory factor in acute respiratory distress syndrome. Nat Med, 1997, 3(3): 320-323. doi:10.1038/nm0397-320. pmid: 9055860
[18]	Bacher M, Metz CN, Calandra T, et al. An essential regulatory role for macrophage migration inhibitory factor in T-cell activation. Proc Natl Acad Sci U S A, 1996, 93(15): 7849-7854. doi:10.1073/pnas.93.15.7849.
[19]	Makita H, Nishimura M, Miyamoto K, et al. Effect of anti-macrophage migration inhibitory factor antibody on lipopolysaccharide-induced pulmonary neutrophil accumulation. Am J Resp Crit Care, 1998, 158(2): 573-579. doi:10.1164/ajrccm.158.2.9707086. pmid: 9700137
[20]	Wang J, Li BX, Ge PP, et al. Mycobacterium tuberculosis suppresses innate immunity by coopting the host ubiquitin system. Nat Immunol, 2015, 16(3): 237-245. doi:10.1038/ni.3096.
[21]	谌蒙蒙, 董静, 孙琦, 等. 基于基因芯片的miRNA表达差异在结核性脑膜炎与病毒性脑膜炎诊断中的价值. 中国防痨杂志, 2022, 44(3): 264-272. doi:10.19982/j.issn.1000-6621.20210699.
[22]	史雨婷, 董静, 贾红彦, 等. miR-99a-5p在结核分枝杆菌感染宿主巨噬细胞中的免疫调控作用. 中国防痨杂志, 2023, 45(5): 464-471. doi:10.19982/j.issn.1000-6621.20220532.
[23]	Mamoori A, Gopalan V, Lu CT, et al. Expression pattern of miR-451 and its target MIF (macrophage migration inhibitory factor) in colorectal cancer. J Clin Pathol, 2017, 70(4): 308-312. doi:10.1136/jclinpath-2016-203972. pmid: 27612504
[24]	Wang MM, Bai Y, Chi HT, et al. miR-451 protects against ischemic stroke by targeting Phd3. Exp Neurol, 2021, 343: 113777. doi:10.1016/j.expneurol.2021.113777.
[25]	Khordadmehr M, Jigari-Asl F, Ezzati H, et al. A comprehensive review on miR-451: A promising cancer biomarker with therapeutic potential. J Cell Physiol, 2019, 234(12): 21716-21731. doi:10.1002/jcp.28888. pmid: 31140618

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

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 1

参考文献 25

相关文章 9

编辑推荐

Metrics

本文评价

友情链接

[1]	史雨婷, 董静, 贾红彦, 朱传智, 杨斌, 李自慧, 孙琦, 杜博平, 邢爱英, 张宗德, 潘丽萍. miR-99a-5p在结核分枝杆菌感染宿主巨噬细胞中的免疫调控作用[J]. 中国防痨杂志, 2023, 45(5): 464-471.
[2]	谌蒙蒙, 董静, 孙琦, 黄麦玲, 丁则昱, 史雨婷, 贾红彦, 杜博平, 魏荣荣, 邢爱英, 张宗德, 潘丽萍. 基于基因芯片的miRNA表达差异在结核性脑膜炎与病毒性脑膜炎诊断中的价值[J]. 中国防痨杂志, 2022, 44(3): 264-272.
[3]	董静, 贾红彦, 孙琦, 李自慧, 魏荣荣, 杜博平, 邢爱英, 潘丽萍, 张宗德. 结核性与恶性胸腔积液中微小核糖核酸内参基因筛选的初步研究[J]. 中国防痨杂志, 2021, 43(3): 261-267.
[4]	刘浩然,张亚莉,任卫聪,赵玲娟,李传友,王伟,高孟秋. 三种细胞因子在肺结核与肺癌胸腔积液中的诊断价值[J]. 中国防痨杂志, 2018, 40(10): 1046-1050.
[5]	央拉，李长山. 肺结核相关microRNA研究进展[J]. 中国防痨杂志, 2015, 37(2): 194-198.
[6]	钟球周琳钱明陈涛陈亮李海成. 结核病患者血清中Small RNAs表达谱分析[J]. 中国防痨杂志, 2011, 33(6): 350-356.
[7]	漆沄石志红苟超伦. MIF、TNFα在肺结核组织中表达和意义的初步探讨[J]. 中国防痨杂志, 2011, 33(3): 153-157.
[8]	张立群孙照刚高孟秋马丽萍吴晓光刘菲. 肺结核患者外周血单核细胞中差异表达miRNA的筛选[J]. 中国防痨杂志, 2011, 33(11): 729-733.
[9]	袁保东,王卫华,段琼红,张炎林,陆兰英. 耐多药肺结核患者生命质量的评价[J]. 中国防痨杂志, 2006, 28(3): 139-142.