MicroRNA‑23a‑3p targeting of HMGB1 inhibits LPS‑induced inflammation in murine macrophages <em>in vitro</em>

Sun,Qi; Wang,Bing; Li,Mengqiu

doi:10.3892/etm.2022.11251

MicroRNA‑23a‑3p targeting of HMGB1 inhibits LPS‑induced inflammation in murine macrophages in vitro

Authors:
- Qi Sun
- Bing Wang
- Mengqiu Li
View Affiliations

Affiliations: Department of Critical Care Medicine, The Jingzhou Central Hospital of Hubei, Jingzhou, Hubei 434000, P.R. China
Published online on: March 10, 2022 https://doi.org/10.3892/etm.2022.11251
Article Number: 322
Copyright: © Sun 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

Inflammatory cytokines, including high mobility group box 1 (HMGB1), play a key role in sepsis via various mechanisms, some of which remain unknown. Sepsis is a common cause of death in patients admitted to the intensive care unit. MicroRNAs (miRs) serve an important role in the inflammatory response. The present study aimed to investigate the role of miR‑23a‑3p in macrophage inflammation and the targeted regulation of HMGB1 expression. The murine macrophage cell line RAW264.7 was subjected to lipopolysaccharide (LPS) treatment to mimic the inflammation involved in sepsis in vitro. Reverse transcription‑quantitative PCR was performed to measure miR‑23a‑3p expression and mRNA expression. Protein levels were determined using ELISA and western blotting. The target binding relationship between miR‑23a‑3p and the HMGB1 3'untranslated region was predicted and validated with a dual luciferase reporter assay. HMGB1 expression was increased and miR‑23a‑3p expression significantly reduced in patients with sepsis and in LPS‑treated RAW264.7 cells in comparison with controls. Overexpression of miR‑23a‑3p reduced interleukin (IL)‑6 and tumor necrosis factor (TNF)‑α expression in RAW264.7 cells under LPS stimulation, while silencing of miR‑23a‑3p elevated the expression of IL‑6 and TNF‑α in comparison with controls. The inhibitory effect of miR‑23a‑3p on LPS‑induced inflammation could be abolished by HMGB1 upregulation in RAW264.7 cells. HMGB1 was targeted by miR‑23a‑3p. miR‑23a‑3p is expressed at reduced levels during inflammation in sepsis, and overexpression of miR‑23a‑3p inhibits LPS‑induced inflammation in murine macrophages in vitro by directly downregulating HMGB1. The results of the present study provided a novel insight into the molecular mechanism underlying HMGB1 expression at the post‑transcriptional level in sepsis.

View Figures

View References

1	Caserta S, Kern F, Cohen J, Drage S, Newbury SF and Llewelyn MJ: Circulating plasma microRNAs can differentiate human sepsis and systemic inflammatory response syndrome (SIRS). Sci Rep. 6(28006)2016.PubMed/NCBI View Article : Google Scholar
2	Tracey KJ: The inflammatory reflex. Nature. 420:853–859. 2002.PubMed/NCBI View Article : Google Scholar
3	Fleischmann C, Scherag A, Adhikari NK, Hartog CS, Tsaganos T, Schlattmann P, Angus DC and Reinhart K: International Forum of Acute Care Trialists. Assessment of global incidence and mortality of Hospital-treated sepsis. Am J Respir Crit Care Mede. 193:259–272. 2016.PubMed/NCBI View Article : Google Scholar
4	Angus DC, Linde-Zwirble WT, Lidicker J, Clermont G, Carcillo J and Pinsky MR: Epidemiology of severe sepsis in the United States: Analysis of incidence, outcome, and associated costs of care. Crit Care Med. 29:1303–1310. 2001.PubMed/NCBI View Article : Google Scholar
5	Cho W, Koo JY, Park Y, Oh K, Lee S, Song JS, Bae MA, Lim D, Lee DS and Park SB: Treatment of sepsis pathogenesis with high mobility group box protein 1-regulating anti-inflammatory agents. J Med Chem. 60:170–179. 2017.PubMed/NCBI View Article : Google Scholar
6	Adib-Conquy M and Cavaillon JM: Stress molecules in sepsis and systemic inflammatory response syndrome. FEBS Lett. 581:3723–3733. 2007.PubMed/NCBI View Article : Google Scholar
7	Lyle NH, Pena OM, Boyd JH and Hancock RE: Barriers to the effective treatment of sepsis: Antimicrobial agents, sepsis definitions, and host-directed therapies. Ann N Y Acad Sci. 1323:101–114. 2014.PubMed/NCBI View Article : Google Scholar
8	Silman NJ: Rapid diagnosis of sepsis using biomarker signatures. Crit Care. 17(1020)2013.PubMed/NCBI View Article : Google Scholar
9	Wang H, Bloom O, Zhang M, Vishnubhakat JM, Ombrellino M, Che J, Frazier A, Yang H, Ivanova S, Borovikova L, et al: HMG-1 as a late mediator of endotoxin lethality in mice. Science. 285:248–251. 1999.PubMed/NCBI View Article : Google Scholar
10	Gonelevue S, Bandyopadhyay A, Bhagat S, Alam MI and Khan GA: Sterile inflammatory role of high mobility group Box 1 protein: Biological functions and involvement in disease. J Vasc Res. 55:244–254. 2018.PubMed/NCBI View Article : Google Scholar
11	Kang R, Chen R, Zhang Q, Hou W, Wu S, Cao L, Huang J, Yu Y, Fan XG, Yan Z, et al: HMGB1 in health and disease. Mol Aspects Med. 40:1–116. 2014.PubMed/NCBI View Article : Google Scholar
12	Magna M and Pisetsky DS: The role of HMGB1 in the pathogenesis of inflammatory and autoimmune diseases. Mol Med. 20:138–146. 2014.PubMed/NCBI View Article : Google Scholar
13	Ulloa L and Tracey KJ: The ʻCytokine Profileʼ: A code for sepsis. Trends Mol Med. 11:56–63. 2005.PubMed/NCBI View Article : Google Scholar
14	Wang H, Yang H and Tracey KJ: Extracellular role of HMGB1 in inflammation and sepsis. J Intern Med. 255:320–331. 2004.PubMed/NCBI View Article : Google Scholar
15	Gibot S, Massin F, Cravoisy A, Barraud D, Nace L, Levy B and Bollaert PE: High-mobility group box 1 protein plasma concentrations during septic shock. Intensive Care Med. 33:1347–1353. 2007.PubMed/NCBI View Article : Google Scholar
16	Andersson U and Tracey KJ: HMGB1 is a therapeutic target for sterile inflammation and infection. Annu Rev Immunol. 29:139–162. 2011.PubMed/NCBI View Article : Google Scholar
17	Kingsley SMK and Bhat BV: Role of microRNAs in sepsis. Inflamm Res. 66:553–569. 2017.PubMed/NCBI View Article : Google Scholar
18	Bulun SE and Nezhat C: Aromatase, microRNA, and inflammation: A complex relationship. Fertil Steril. 106:552–553. 2016.PubMed/NCBI View Article : Google Scholar
19	Essandoh K and Fan GC: Role of extracellular and intracellular microRNAs in sepsis. Biochim Biophys Acta. 1842:2155–2162. 2014.PubMed/NCBI View Article : Google Scholar
20	Yao Y, Sun F and Lei M: miR-25 inhibits sepsis-induced cardiomyocyte apoptosis by targetting PTEN. Biosci Rep. 38(BSR20171511)2018.PubMed/NCBI View Article : Google Scholar
21	Ma Y, Liu Y, Hou H, Yao Y and Meng H: MiR-150 predicts survival in patients with sepsis and inhibits LPS-induced inflammatory factors and apoptosis by targeting NF-κB1 in human umbilical vein endothelial cells. Biochem Biophys Res Commun. 500:828–837. 2018.PubMed/NCBI View Article : Google Scholar
22	Benz F, Roy S, Trautwein C, Roderburg C and Luedde T: Circulating MicroRNAs as biomarkers for sepsis. Int J Mol Sci. 17(78)2016.PubMed/NCBI View Article : Google Scholar
23	Chhabra R, Dubey R and Saini N: Cooperative and individualistic functions of the microRNAs in the miR-23a~27a~24-2 cluster and its implication in human diseases. Mol Cancer. 9(232)2010.PubMed/NCBI View Article : Google Scholar
24	Gusar VA, Timofeeva AV, Zhanin IS, Shram SI and Pinelis VG: Estimation of Time-dependent microRNA expression patterns in brain tissue, leukocytes, and blood plasma of rats under photochemically induced focal cerebral ischemia. Mol Biol (Mosk). 51:683–695. 2017.PubMed/NCBI View Article : Google Scholar : (In Russian).
25	Lozano-Bartolomé J, Llauradó G, Portero-Otin M, Altuna-Coy A, Rojo-Martinez G, Vendrell J, Jorba R, Rodríguez-Gallego E and Chacón MR: Altered expression of miR-181a-5p and miR-23a-3p is associated with obesity and TNFα-induced insulin resistance. J Clin Endocrinol Metab. 103:1447–1458. 2018.PubMed/NCBI View Article : Google Scholar
26	Parker MI and Palladino MA: MicroRNAs downregulated following immune activation of rat testis. Am J Reprod Immunol: 77, 2017 doi: 10.1111/aji.12673.
27	Ge QM, Huang CM, Zhu XY, Bian F and Pan SM: Differentially expressed miRNAs in sepsis-induced acute kidney injury target oxidative stress and mitochondrial dysfunction pathways. PLoS One. 12(e0173292)2017.PubMed/NCBI View Article : Google Scholar
28	Chen X, Liu Y, Gao Y, Shou S and Chai Y: The roles of macrophage polarization in the host immune response to sepsis. Int Immunopharmacoly. 96(107791)2021.PubMed/NCBI View Article : Google Scholar
29	Zhou W, Wang J, Li Z, Li J and Sang M: MicroRNA-2055b inhibits HMGB1 expression in LPS-induced sepsis. Int J Mol Med. 38:312–318. 2016.PubMed/NCBI View Article : Google Scholar
30	Singer M, Deutschman CS, Seymour CW, Shankar-Hari M, Annane D, Bauer M, Bellomo R, Bernard GR, Chiche JD, Coopersmith CM, et al: The third international consensus definitions for sepsis and septic shock (Sepsis-3). JAMA. 315:801–810. 2016.PubMed/NCBI View Article : Google Scholar
31	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.PubMed/NCBI View Article : Google Scholar
32	Chen W, Ma X, Zhang P, Li Q, Liang X and Liu J: miR-212-3p inhibits LPS-induced inflammatory response through targeting HMGB1 in murine macrophages. Exp Cell Res. 350:318–326. 2017.PubMed/NCBI View Article : Google Scholar
33	Paraskevopoulou MD, Georgakilas G, Kostoulas N, Vlachos IS, Vergoulis T, Reczko M, Filippidis C, Dalamagas T and Hatzigeorgiou AG: DIANA-microT web server v5.0: Service integration into miRNA functional analysis workflows. Nucleic Acids Res. 41:W169–W173. 2013.PubMed/NCBI View Article : Google Scholar
34	Stevens NE, Chapman MJ, Fraser CK, Kuchel TR, Hayball JD and Diener KR: Therapeutic targeting of HMGB1 during experimental sepsis modulates the inflammatory cytokine profile to one associated with improved clinical outcomes. Sci Rep. 7(5850)2017.PubMed/NCBI View Article : Google Scholar
35	Yang H, Ochani M, Li J, Qiang X, Tanovic M, Harris HE, Susarla SM, Ulloa L, Wang H, DiRaimo R, et al: Reversing established sepsis with antagonists of endogenous high-mobility group box 1. Proc Natl Acad Sci USA. 101:296–301. 2004.PubMed/NCBI View Article : Google Scholar
36	Charoensup J, Sermswan RW, Paeyao A, Promakhejohn S, Punasee S, Chularari C, Krabkraikaew S, Lertanekawattana S and Wongratanacheewin S: High HMGB1 level is associated with poor outcome of septicemic melioidosis. Int J Infect Dis. 28:111–116. 2014.PubMed/NCBI View Article : Google Scholar
37	Guo ZS, Liu Z, Bartlett DL, Tang D and Lotze MT: Life after death: Targeting high mobility group box 1 in emergent cancer therapies. Am J Cancer Res. 3:1–20. 2013.PubMed/NCBI
38	Andersson U, Yang H and Harris H: Extracellular HMGB1 as a therapeutic target in inflammatory diseases. Expert Opin Ther Targets. 22:263–277. 2018.PubMed/NCBI View Article : Google Scholar
39	Deng M, Tang Y, Li W, Wang X, Zhang R, Zhang X, Zhao X, Liu J, Tang C, Liu Z, et al: The endotoxin delivery protein HMGB1 mediates Caspase-11-dependent lethality in sepsis. Immunity. 49:740–753.e7. 2018.PubMed/NCBI View Article : Google Scholar
40	Kim HM and Kim YM: HMGB1: LPS delivery vehicle for caspase-11-mediated pyroptosis. Immunity. 49:582–584. 2018.PubMed/NCBI View Article : Google Scholar
41	Wu D, Pan P, Su X, Zhang L, Qin Q, Tan H, Huang L and Li Y: Interferon regulatory factor-1 mediates alveolar macrophage pyroptosis during LPS-induced acute lung injury in mice. Shock. 46:329–338. 2016.PubMed/NCBI View Article : Google Scholar
42	Yuan Z, Luo G, Li X, Chen J, Wu J and Peng Y: PPARγ inhibits HMGB1 expression through upregulation of miR-142-3p in vitro and in vivo. Cell Signal. 28:158–164. 2016.PubMed/NCBI View Article : Google Scholar
43	Park EJ, Kim YM, Kim HJ and Chang KC: Degradation of histone deacetylase 4 via the TLR4/JAK/STAT1 signaling pathway promotes the acetylation of high mobility group box 1 (HMGB1) in lipopolysaccharide-activated macrophages. FEBS Open Bio. 8:1119–1126. 2018.PubMed/NCBI View Article : Google Scholar
44	Lee W, Ku SK and Bae JS: Zingerone reduces HMGB1-mediated septic responses and improves survival in septic mice. Toxicol Appl Pharmacol. 329:202–211. 2017.PubMed/NCBI View Article : Google Scholar
45	Quan J, Pan X, Li Y, Hu Y, Tao L, Li Z, Zhao L, Wang J, Li H, Lai Y, et al: miR-23a-3p acts as an oncogene and potential prognostic biomarker by targeting PNRC2 in RCC. Biomed Pharmacother. 110:656–666. 2018.PubMed/NCBI View Article : Google Scholar
46	Zhao H, Tao Z, Wang R, Liu P, Yan F, Li J, Zhang C, Ji X and Luo Y: MicroRNA-23a-3p attenuates oxidative stress injury in a mouse model of focal cerebral ischemia-reperfusion. Brain Res. 1592:65–72. 2014.PubMed/NCBI View Article : Google Scholar
47	Minnich DJ and Moldawer LL: Anti-cytokine and anti-inflammatory therapies for the treatment of severe sepsis: Progress and pitfalls. Proc Nutr Soc. 63:437–441. 2004.PubMed/NCBI View Article : Google Scholar