MicroRNA‑502‑3p promotes <em>Mycobacterium tuberculosis</em> survival in macrophages by modulating the inflammatory response by targeting ROCK1

Liu,Fang; Dong,Zhen; Lin,Yuefu; Yang,Haibo; Wang,Pingping; Zhang,Yongxia

doi:10.3892/mmr.2021.12393

MicroRNA‑502‑3p promotes Mycobacterium tuberculosis survival in macrophages by modulating the inflammatory response by targeting ROCK1

Authors:
- Fang Liu
- Zhen Dong
- Yuefu Lin
- Haibo Yang
- Pingping Wang
- Yongxia Zhang
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Affiliations: Respiratory Endoscopy Room, Linyi People's Hospital, Linyi, Shandong 276034, P.R. China, East Medical District Office, Linyi People's Hospital, Linyi, Shandong 276034, P.R. China, Department of Prevention, Linyi People's Hospital, Linyi, Shandong 276034, P.R. China, Department of Occupational Diseases, Linyi People's Hospital, Linyi, Shandong 276034, P.R. China, Rehabilitation Department, Shandong Coal Linyi Hot Spring Sanatorium, Linyi, Shandong 276034, P.R. China, Emergency Department, Linyi People's Hospital, Linyi, Shandong 276034, P.R. China
Published online on: September 1, 2021 https://doi.org/10.3892/mmr.2021.12393
Article Number: 753
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Tuberculosis (TB) is caused by Mycobacterium tuberculosis (M. tuberculosis) infection and has the highest mortality rate of any single infectious disease worldwide. The aim of the present study was to investigate the function of microRNA (miR)‑502‑3p in M. tuberculosis‑infected macrophages. The Gene Expression Omnibus database was used to analyze miR‑502‑3p expression in patients with TB and healthy individuals. THP‑1 and RAW 264.7 cells were transfected with miR‑502‑3p mimic, miR‑502‑3p inhibitor, pcDNA3.1‑ROCK1 or their negative controls. The expression levels of miR‑502‑3p and inflammatory cytokines were evaluated using reverse transcription‑quantitative PCR. The colony‑forming unit assay was performed to assess the survival of M. tuberculosis in macrophages, and Toll‑like receptor (TLR)4/NF‑κB signaling pathway‑associated protein expression levels were detected by western blotting. The nuclear translocation of NF‑κB p65 was detected via immunocytochemistry. TargetScan was used to predict the binding sites between miR‑502‑3p and ROCK1. The interaction between miR‑502‑3p and Rho‑associated coiled‑coil‑forming protein kinase 1 (ROCK1) was confirmed using a dual‑luciferase reporter assay; ROCK1 was demonstrated to be a direct target gene of miR‑502‑3p. Results from the present study demonstrated that miR‑502‑3p expression was significantly increased during M. tuberculosis infection in macrophages. Upregulation of miR‑502‑3p expression levels significantly enhanced the survival of intracellular M. tuberculosis. IL‑6, TNF‑α, and IL‑1β mRNA expression levels were significantly upregulated during M. tuberculosis infection but were downregulated by miR‑502‑3p overexpression. Moreover, miR‑502‑3p mimics transfection significantly downregulated TLR4/NF‑κB signaling pathway‑associated protein expression and significantly reduced nuclear transcription of NF‑κB in M. tuberculosis‑infected macrophages. ROCK1 overexpression reversed the miR‑502‑3p inhibitory effect on cytokine production in M. tuberculosis‑infected macrophages. In conclusion, miR‑502‑3p/ROCK1 may serve an anti‑inflammatory role and may improve the survival of M. tuberculosis within macrophages, which may provide a promising therapeutic target for TB.

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1	Alzahabi KH, Usmani O, Georgiou TK, Ryan MP, Robertson BD, Tetley TD and Porter AE: Approaches to treating tuberculosis by encapsulating metal ions and anti-mycobacterial drugs utilizing nano- and microparticle technologies. Emerg Top Life Sci. 4:581–600. 2020. View Article : Google Scholar : PubMed/NCBI
2	Harding E: WHO global progress report on tuberculosis elimination. Lancet Respir Med. 8:30418–7. 2020. View Article : Google Scholar
3	Koch A and Mizrahi V: Mycobacterium tuberculosis. Trends Microbiol. 26:555–556. 2018. View Article : Google Scholar : PubMed/NCBI
4	Leopold Wager CM, Arnett E and Schlesinger LS: Mycobacterium tuberculosis and macrophage nuclear receptors: What we do and don't know. Tuberculosis (Edinb). Apr 25–2019.(Epub ahead of print). doi: 10.1016/j.tube.2019.04.016. View Article : Google Scholar : PubMed/NCBI
5	Correia de Sousa M, Gjorgjieva M, Dolicka D, Sobolewski C and Foti M: Deciphering miRNAs' Action through miRNA Editing. Int J Mol Sci. 20:62492019. View Article : Google Scholar : PubMed/NCBI
6	Zhang X, Zhu M and Hu X: Integrated miRNA and mRNA expression profiling to identify mRNA targets of dysregulated miRNAs in pulmonary tuberculosis. Epigenomics. 10:1051–1069. 2018. View Article : Google Scholar : PubMed/NCBI
7	Fu B, Xue W, Zhang H, Zhang R, Feldman K, Zhao Q, Zhang S, Shi L, Pavani KC, Nian W, et al: MicroRNA-325-3p facilitates immune escape of Mycobacterium tuberculosis through targeting LNX1 via NEK6 accumulation to promote anti-apoptotic STAT3 signaling. mBio. 11:00557–20. 2020. View Article : Google Scholar : PubMed/NCBI
8	Sahu SK, Kumar M, Chakraborty S, Banerjee SK, Kumar R, Gupta P, Jana K, Gupta UD, Ghosh Z, Kundu M and Basu J: MicroRNA 26a (miR-26a)/KLF4 and CREB-C/EBPβ regulate innate immune signaling, the polarization of macrophages and the trafficking of Mycobacterium tuberculosis to lysosomes during infection. PLoS Pathog. 13:e10064102017. View Article : Google Scholar : PubMed/NCBI
9	Niu W, Sun B, Li M, Cui J, Huang J and Zhang L: TLR-4/microRNA-125a/NF-κB signaling modulates the immune response to Mycobacterium tuberculosis infection. Cell Cycle. 17:1931–1945. 2018. View Article : Google Scholar : PubMed/NCBI
10	Chen LY, Zuraw BL, Liu FT, Huang S and Pan ZK: IL-1 receptor-associated kinase and low molecular weight GTPase RhoA signal molecules are required for bacterial lipopolysaccharide-induced cytokine gene transcription. J Immunol. 169:3934–3939. 2002. View Article : Google Scholar : PubMed/NCBI
11	Shimizu S, Tahara M, Ogata S, Hashimoto K, Morishige K, Tasaka K and Murata Y: Involvement of nuclear factor-kB activation through RhoA/Rho-kinase pathway in LPS-induced IL-8 production in human cervical stromal cells. Mol Hum Reprod. 13:181–187. 2007. View Article : Google Scholar : PubMed/NCBI
12	Wang ZH, Zhu D, Xie S, Deng Y, Pan Y, Ren J and Liu HG: Inhibition of Rho-kinase attenuates left ventricular remodeling caused by chronic intermittent hypoxia in rats via suppressing myocardial inflammation and apoptosis. J Cardiovasc Pharmacol. 70:102–109. 2017. View Article : Google Scholar : PubMed/NCBI
13	Juarez E, Nuñez C, Sada E, Ellner JJ, Schwander SK and Torres M: Differential expression of Toll-like receptors on human alveolar macrophages and autologous peripheral monocytes. Respir Res. 11:1465–9921. 2010. View Article : Google Scholar : PubMed/NCBI
14	Guo H, Zhang Y, Cheng BC, Fu X, Zhu P, Chen J, Chan Y, Yin C, Wang Y, Hossen M, et al: An ethanolic extract of the aerial part of Siegesbeckia orientalis L. inhibits the production of inflammatory mediators regulated by AP-1, NF-κB and IRF3 in LPS-stimulated RAW 264.7 cells. Biosci Trends. 12:330–337. 2018. View Article : Google Scholar : PubMed/NCBI
15	Sierra-Mondragon E, Molina-Jijon E, Namorado-Tonix C, Rodríguez-Muñoz R, Pedraza-Chaverri J and Reyes JL: All-trans retinoic acid ameliorates inflammatory response mediated by TLR4/NF-κB during initiation of diabetic nephropathy. J Nutr Biochem. 60:47–60. 2018. View Article : Google Scholar : PubMed/NCBI
16	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
17	Korbel DS, Schneider BE and Schaible UE: Innate immunity in tuberculosis: Myths and truth. Microbes Infect. 10:995–1004. 2008. View Article : Google Scholar : PubMed/NCBI
18	Sharma N, Verma R, Kumawat KL, Basu A and Singh SK: miR-146a suppresses cellular immune response during Japanese encephalitis virus JaOArS982 strain infection in human microglial cells. J Neuroinflammation. 12:302015. View Article : Google Scholar : PubMed/NCBI
19	Pachathundikandi SK and Backert S: Helicobacter pylori controls NLRP3 expression by regulating hsa-miR-223-3p and IL-10 in cultured and primary human immune cells. Innate Immun. 24:11–23. 2018. View Article : Google Scholar : PubMed/NCBI
20	Yu T, Ju Z, Luo M, Hu R, Teng Y, Xie L, Zhong C, Chen L, Hou W, Xiong Y and Feng Y: Elevated expression of miR-146a correlates with high levels of immune cell exhaustion markers and suppresses cellular immune function in chronic HIV-1-infected patients. Sci Rep. 9:188292019. View Article : Google Scholar : PubMed/NCBI
21	Cui J, Li Z, Cui K, Gao Y, Zhang B, Niu J and Wang Y: MicroRNA-20a-3p regulates the host immune response to facilitate the Mycobacterium tuberculosis infection by targeting IKKβ/NF-κB pathway. Int Immunopharmacol. 91:1072862021. View Article : Google Scholar : PubMed/NCBI
22	Jin H, Yu M, Lin Y, Hou B, Wu Z, Li Z and Sun J: MiR-502-3P suppresses cell proliferation, migration, and invasion in hepatocellular carcinoma by targeting SET. Onco Targets Ther. 9:3281–3289. 2016.PubMed/NCBI
23	Zhang J, Hou L, Liang R, Chen X, Zhang R, Chen W and Zhu J: CircDLST promotes the tumorigenesis and metastasis of gastric cancer by sponging miR-502-5p and activating the NRAS/MEK1/ERK1/2 signaling. Mol Cancer. 18:802019. View Article : Google Scholar : PubMed/NCBI
24	Specjalski K, Maciejewska A, Pawłowski R, Chełmińska M and Jassem E: Changes in the expression of microRNA in the buildup phase of wasp venom immunotherapy: A pilot study. Int Arch Allergy Immunol. 170:97–100. 2016. View Article : Google Scholar : PubMed/NCBI
25	Lv J, He X, Wang H, Wang Z, Kelly GT, Wang X, Chen Y, Wang T and Qian Z: TLR4-NOX2 axis regulates the phagocytosis and killing of Mycobacterium tuberculosis by macrophages. BMC Pulm Med. 17:1942017. View Article : Google Scholar : PubMed/NCBI
26	Shi L, Jiang Q, Bushkin Y, Subbian S and Tyagi S: Biphasic dynamics of macrophage immunometabolism during Mycobacterium tuberculosis infection. mBio. 10:e02550–18. 2019. View Article : Google Scholar : PubMed/NCBI
27	Li X, Huang S, Yu T, Liang G, Liu H, Pu D and Peng N: MIR-140 modulates the inflammatory responses of Mycobacterium tuberculosis-infected macrophages by targeting TRAF6. J Cell Mol Med. 23:5642–5653. 2019. View Article : Google Scholar : PubMed/NCBI
28	Gu X, Gao Y, Mu DG and Fu EQ: MiR-23a-5p modulates mycobacterial survival and autophagy during Mycobacterium tuberculosis infection through TLR2/MyD88/NF-κB pathway by targeting TLR2. Exp Cell Res. 354:71–77. 2017. View Article : Google Scholar : PubMed/NCBI
29	Zhao Z, Hao J, Li X, Chen Y and Qi X: MiR-21-5p regulates mycobacterial survival and inflammatory responses by targeting Bcl-2 and TLR4 in Mycobacterium tuberculosis-infected macrophages. FEBS Lett. 593:1326–1335. 2019. View Article : Google Scholar : PubMed/NCBI
30	Liu HY: Down-regulation of miR-144 after Mycobacterium tuberculosis infection promotes inflammatory factor secretion from macrophages through the Tpl2/ERK pathway. Cell Mol Biol. 62:87–93. 2016.PubMed/NCBI
31	Gong J, Guan L, Tian P, Li C and Zhang Y: Rho kinase type 1 (ROCK1) promotes lipopolysaccharide-induced inflammation in corneal epithelial cells by activating toll-like receptor 4 (TLR4)-mediated signaling. Med Sci Monit. 24:3514–3523. 2018. View Article : Google Scholar : PubMed/NCBI
32	Chopra P, Koduri H, Singh R, Koul A, Ghildiyal M, Sharma K, Tyagi AK and Singh Y: Nucleoside diphosphate kinase of Mycobacterium tuberculosis acts as GTPase-activating protein for Rho-GTPases. FEBS Lett. 571:212–216. 2004. View Article : Google Scholar : PubMed/NCBI
33	Wu H, Bao Y, Wang L, Li X and Sun J: Mycobacterium marinum down-regulates miR-148a in macrophages in an EsxA-dependent manner. Int Immunopharmacol. 73:41–48. 2019. View Article : Google Scholar : PubMed/NCBI