MicroRNA‑146a inhibits NF‑κB activation and pro‑inflammatory cytokine production by regulating IRAK1 expression in THP‑1 cells

Zhou,Chunlei; Zhao,Lan; Wang,Kai; Qi,Qianru; Wang,Meng; Yang,Lei; Sun,Ping; Mu,Hong

doi:10.3892/etm.2019.7881

MicroRNA‑146a inhibits NF‑κB activation and pro‑inflammatory cytokine production by regulating IRAK1 expression in THP‑1 cells

Authors:
- Chunlei Zhou
- Lan Zhao
- Kai Wang
- Qianru Qi
- Meng Wang
- Lei Yang
- Ping Sun
- Hong Mu
View Affiliations

Affiliations: Department of Medical Laboratory, Tianjin First Central Hospital, Tianjin 300192, P.R. China, Laboratory of Acupuncture and Moxibustion, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300193, P.R. China, Department of Medical Laboratory, Tianjin Children's Hospital, Tianjin 300134, P.R. China
Published online on: August 13, 2019 https://doi.org/10.3892/etm.2019.7881
Pages: 3078-3084
Copyright: © Zhou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

MicroRNA (miR)‑146a levels are reduced in peripheral blood mononuclear cells of patients with systemic lupus erythematosus (SLE); however, its function is not well understood. The present study investigated the role of miR‑146a in the regulation of lipopolysaccharide (LPS)‑induced inflammation in THP‑1 cells. A miR‑146a mimic and an inhibitor were used to overexpress and downregulate miR‑146a expression, respectively. Reverse transcription‑quantitative PCR and western blot analyses were performed to evaluate interleukin (IL)‑1 receptor‑associated kinase 1 (IRAK1) expression, and western blot analysis was applied to assess nuclear factor‑κB activation by analyzing p65 subunit levels in the nucleus. To investigate the effects of miR‑146a on LPS‑induced inflammation, IL‑6 and tumor necrosis factor‑α (TNF‑α) levels were also measured using ELISA. The results of the present study revealed thatmiR‑146a overexpression significantly reduced IRAK1 expression, reduced p65 levels in the nucleus and reduced IL‑6 and TNF‑α levels in the supernatant of the cell culture medium of THP‑1 cells following LPS treatment. Luciferase assays confirmed IRAK1 to be a direct target of miR‑146a in THP‑1 cells. In conclusion, miR‑146a may regulate IRAK1 expression and inhibit the activation of inflammatory signals and secretion of pro‑inflammatory cytokines. The present study revealed, at least in part, the mechanisms by which miR‑146a regulate the inflammatory response in SLE.

View Figures

View References

1	Ranganathan K and Sivasankar V: MicroRNAs-biology and clinical applications. J Oral Maxillofac Pathol. 18:229–234. 2014. View Article : Google Scholar : PubMed/NCBI
2	Pileczki V, Cojocneanu-Petric R, Maralani M, Neagoe IB and Sandulescu R: MicroRNAs as regulators of apoptosis mechanisms in cancer. Clujul Med. 89:50–55. 2016.PubMed/NCBI
3	Miao CG, Yang YY, He X, Huang C, Huang Y, Zhang L, Lv XW, Jin Y and Li J: The emerging role of microRNAs in the pathogenesis of systemic lupus erythematosus. Cell Signal. 25:1828–1836. 2013. View Article : Google Scholar : PubMed/NCBI
4	Sharma AR, Sharma G, Lee SS and Chakraborty C: MiRNA-regulated key components of cytokine signaling pathways and inflammation in rheumatoid arthritis. Med Res Rev. 36:425–439. 2016. View Article : Google Scholar : PubMed/NCBI
5	Habibi F, Ghadiri Soufi F, Ghiasi R, Khamaneh AM and Alipour MR: Alteration in inflammation-related miR-146a expression in NF-κB signaling pathway in diabetic rat hippocampus. Adv Pharm Bull. 6:99–103. 2016. View Article : Google Scholar : PubMed/NCBI
6	Stickel N, Prinz G, Pfeifer D, Hasselblatt P, Schmitt-Graeff A, Follo M, Thimme R, Finke J, Duyster J, Salzer U and Zeiser R: MiR-146a regulates the TRAF6/TNF-axis in donor T cells during GVHD. Blood. 124:2586–2595. 2014. View Article : Google Scholar : PubMed/NCBI
7	Taganov KD, Boldin MP and Baltimore D: MicroRNAs and immunity: Tiny players in a big field. Immunity. 26:133–137. 2007. View Article : Google Scholar : PubMed/NCBI
8	Perl A: Pathogenic mechanisms in systemic lupus erythematosus. Autoimmunity. 43:1–6. 2010. View Article : Google Scholar : PubMed/NCBI
9	Wang H, Peng W, Ouyang X, Li W and Dai Y: Circulating microRNAs as candidate biomarkers in patients with systemic lupus erythematosus. Transl Res. 160:198–206. 2012. View Article : Google Scholar : PubMed/NCBI
10	Ji JD, Cha ES and Lee WJ: Association of miR-146a polymorphisms with systemic lupus erythematosus: A meta-analysis. Lupus. 23:1023–1030. 2014. View Article : Google Scholar : PubMed/NCBI
11	Gao M, Wang X, Zhang X, Ha T, Ma H, Liu L, Kalbfleisch JH, Gao X, Kao RL, Williams DL and Li C: Attenuation of cardiac dysfunction in polymicrobial sepsis by MicroRNA-146a is mediated via targeting of IRAK1 and TRAF6 expression. J Immunol. 195:672–682. 2015. View Article : Google Scholar : PubMed/NCBI
12	Kaisho T and Aldra S: Toll-like receptor function and signaling. J Allergy Clin Immunol. 117:979–988. 2006. View Article : Google Scholar : PubMed/NCBI
13	Rodriguez-Pla A, Patel P, Maecker HT, Rossello-Urgell J, Baldwin N, Bennett L, Cantrell V, Baisch J, Punaro M, Gotte A, et al: IFN priming is necessary but not sufficient to turn on a migratory dendritic cell program in lupus monocytes. J Immunol. 192:5586–5598. 2014. View Article : Google Scholar : PubMed/NCBI
14	Wang P, Han X, Mo B, Huang G and Wang C: LPS enhances TLR4 expression and IFN-γ production via the TLR4/IRAK/NF-κB signaling pathway in rat pulmonary arterial smooth muscle cells. Mol Med Rep. 16:3111–3116. 2017. View Article : Google Scholar : PubMed/NCBI
15	Nahid MA, Pauley KM, Satoh M and Chan EK: MiR-146a is critical for endotoxin-induced tolerance: Implication in innate immunity. J Biol Chem. 284:34590–34599. 2009. 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	Chen Y, Zeng Z, Shen X, Wu Z, Dong Y and Cheng JC: MicroRNA-146a-5p negatively regulates pro-inflammatory cytokine secretion and cell activation in lipopolysaccharide stimulated human hepatic stellate cells through inhibition of Toll-like receptor 4 signaling pathways. Int J Mol Sci1. 17(pii): E10762016. View Article : Google Scholar
18	Muñoz LE, Janko C, Schulze C, Schorn C, Sarter K, Schett G and Herrmann M: Autoimmunity and chronic inflammation-two clearance-related steps in the etiopathogenesis of SLE. Autoimmun Rev. 10:38–42. 2010. View Article : Google Scholar : PubMed/NCBI
19	Dai Y, Huang YS, Tang M, Lv TY, Hu CX, Tan YH, Xu ZM and Yin YB: Microarray analysis of microRNA expression in peripheral blood cells of systemic lupus erythematosus patients. Lupus. 16:939–946. 2007. View Article : Google Scholar : PubMed/NCBI
20	Tang Y, Luo X, Cui H, Ni X, Yuan M, Guo Y, Huang X, Zhou H, de Vries N, Tak PP, et al: MicroRNA-146A contributes to abnormal activation of the type I interferon pathway in human lupus by targeting the key signaling proteins. Arthritis Rheum. 60:1065–1075. 2009. View Article : Google Scholar : PubMed/NCBI
21	Wigren M, Nilsson J and Kaplan MJ: Pathogenic immunity in systemic lupus erythematosus and atherosclerosis: Common mechanisms and possible targets for intervention. J Intern Med. 278:494–506. 2015. View Article : Google Scholar : PubMed/NCBI
22	Means TK, Latz E, Hayashi F, Murali MR, Golenbock DT and Luster AD: Human lupus autoantibody-DNA complexes activate DCs through cooperation of CD32 and TLR9. J Clin Invest. 115:407–417. 2005. View Article : Google Scholar : PubMed/NCBI
23	Husakova M: MicroRNAs in the key events of systemic lupus erythematosus pathogenesis. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 160:327–342. 2016. View Article : Google Scholar : PubMed/NCBI
24	Fang H, Wang PF, Zhou Y, Wang YC and Yang QW: Toll-like receptor 4 signaling in intracerebral hemorrhage-induced inflammation and injury. J Neuroinflammation. 10:272013. View Article : Google Scholar : PubMed/NCBI
25	He X, Zheng Y, Liu S, Shi S, Liu Y, He Y, Zhang C and Zhou X: MiR-146a protects small intestine against ischemia/reperfusion injury by down-regulatingTLR4/TRAF6/NF-κB pathway. J Cell Physiol. 233:2476–2488. 2018. View Article : Google Scholar : PubMed/NCBI
26	Nahid MA, Satoh M and Chan EK: MicroRNA in TLR signaling and endotoxin tolerance. Cell Mol Immunol. 8:388–403. 2011. View Article : Google Scholar : PubMed/NCBI
27	Willis R, Seif AM, McGwin G Jr, Martinez-Martinez LA, González EB, Dang N, Papalardo E, Liu J, Vilá LM, Reveille JD, et al: Effect of hydroxychloroquine treatment on pro-inflammatory cytokines and disease activity in SLE patients: Data from LUMINA (LXXV), a multiethnic US cohort. Lupus. 21:830–835. 2012. View Article : Google Scholar : PubMed/NCBI
28	Ripley BJ, Goncalves B, Isenberg DA, Latchman DS and Rahman A: Raised levels of interleukin 6 in systemic lupus erythematosus correlate with anaemia. Ann Rheum Dis. 64:849–853. 2005. View Article : Google Scholar : PubMed/NCBI
29	Sabry A, Sheashaa H, EI-Husseini A, Mahmoud K, Eldahshan KF, George SK, Abdel-Khalek E, El-Shafey EM and Abo-Zenah H: Proinflammatory cytokines (TNF-alpha and IL-6) in Egyptian patients with SLE: Its correlation with disease activity. Cytokine. 35:148–153. 2006. View Article : Google Scholar : PubMed/NCBI
30	Arora V, Verma J, Marwah V, Kumar A, Anand D and Das N: Cytokine imbalance in systemic lupus erythematosus: A study on northern Indian subjects. Lupus. 21:596–603. 2012. View Article : Google Scholar : PubMed/NCBI
31	Gigante A, Gasperini ML, Afeltra A, Barbano B, Margiotta D, Cianci R, De Francesco I and Amoroso A: Cytokines expression in SLE nephritis. Eur Rev Med Pharmacol Sci. 15:15–24. 2011.PubMed/NCBI
32	Robinson ES and Werth VP: The role of cytokines in the pathogenesis of cutaneous lupus erythematosus. Cytokine. 73:326–334. 2015. View Article : Google Scholar : PubMed/NCBI
33	Nahid MA, Satoh M and Chan EK: Mechanistic role of microRNA-146a in endotoxin-induced differential cross-regulation of TLR signaling. J Immunol. 186:1723–1734. 2011. View Article : Google Scholar : PubMed/NCBI
34	Du L, Chen X, Duan Z, Liu C, Zeng R, Chen Q and Li M: MiR-146a negatively regulatesdectin-1-induced inflammatory responses. Oncotarget. 8:37355–37366. 2017. View Article : Google Scholar : PubMed/NCBI
35	Mookherjee N and El-Gabalawy HS: High degree of correlation between whole blood and PBMC expression levels of miR-155 and miR-146a in healthy controls and rheumatoid arthritis patients. J Immunol Methods. 400-401:106–110. 2013. View Article : Google Scholar : PubMed/NCBI
36	Sheppard HM, Verdon D, Brooks AE, Feisst V, Ho YY, Lorenz N, Fan V, Birch NP, Didsbury A and Dunbar PR: MicroRNA regulation in human CD8+ T cell subsets-cytokine exposure alone drives miR-146a expression. J Transl Med. 12:2922014. View Article : Google Scholar : PubMed/NCBI
37	Cho S, Lee HM, Yu IS, Choi YS, Huang HY, Hashemifar SS, Lin LL, Chen MC, Afanasiev ND, Khan AA, et al: Differential cell-intrinsic regulations of germinal center B and T cells by miR-146a and miR-146b. Nat Commun. 9:27572018. View Article : Google Scholar : PubMed/NCBI
38	Liu AM, Wang W and Luk JM: miRNAs: New tools for molecular classification, diagnosis and prognosis of hepatocellular carcinoma. Hepat Oncol. 1:323–329. 2014. View Article : Google Scholar : PubMed/NCBI