Anti‑inflammatory effects of miR‑150 are associated with the downregulation of STAT1 in macrophages following lipopolysaccharide treatment

Chen,Song; Zhu,Haijun; Sun,Jie; Zhu,Lili; Qin,Long; Wan,Jian

doi:10.3892/etm.2021.10483

Anti‑inflammatory effects of miR‑150 are associated with the downregulation of STAT1 in macrophages following lipopolysaccharide treatment

Authors:
- Song Chen
- Haijun Zhu
- Jie Sun
- Lili Zhu
- Long Qin
- Jian Wan
View Affiliations

Affiliations: Department of Emergency and Critical Care Medicine, The People's Hospital of Pudong New Area, Shanghai University of Health and Science, Shanghai 201200, P.R. China
Published online on: July 23, 2021 https://doi.org/10.3892/etm.2021.10483
Article Number: 1049
Copyright: © Chen 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

Sepsis is a condition that is associated with high rates of mortality. It is characterized by serious systemic inflammatory responses induced by pathogenic invasion. Although microRNA‑150 (miR‑150) has been previously reported to be involved in the modulation of sepsis, the underlying molecular mechanism in sepsis remains poorly understood. In the present study, the human monocytic cell line THP‑1 was treated with LPS to mimic sepsis in vitro, following which miR‑150 and STAT1 expression were measured using reverse transcription‑quantitative PCR or western blotting. Secretion of inflammatory cytokines interleukin (IL)‑1β, IL‑6 and tumor necrosis factor‑α (TNF‑α) into the medium were measured by ELISA. The potential relationship between STAT1 and miR‑150 was determined using dual‑luciferase reporter and RNA immunoprecipitation assays. miR‑150 expression was found to be was downregulated by LPS treatment in THP‑1 cells in both dose‑ and time‑dependent manners. LPS treatment also induced IL‑1β, IL‑6 and TNF‑α secretion in a manner that could be inhibited by miR‑150 overexpression and enhanced by transfection with the miR‑150 inhibitor. miR‑150 was revealed to directly target STAT1 by negatively regulating its expression. In addition, STAT1 expression was demonstrated to be upregulated by LPS treatment. STAT1 overexpression reversed the inhibitory effects of miR‑150 overexpression on IL‑1β, IL‑6 and TNF‑α secretion whilst STAT1 knockdown attenuated IL‑1β, IL‑6 and TNF‑α secretion induced by miR‑150 inhibitor transfection. In conclusion, the present study suggested that miR‑150 regulates the inflammatory response in macrophages following LPS challenge by regulating the expression of STAT1.

View Figures

View References

1	Csoka B, Nemeth ZH, Toro G, Idzko M, Zech A, Koscso B, Spolarics Z, Antonioli L, Cseri K, Erdelyi K, et al: Extracellular ATP protects against sepsis through macrophage P2X7 purinergic receptors by enhancing intracellular bacterial killing. FASEB J. 29:3626–3637. 2015.PubMed/NCBI View Article : Google Scholar
2	Rudd KE, Johnson SC, Agesa KM, Shackelford KA, Tsoi D, Kievlan DR, Colombara DV, Ikuta KS, Kissoon N, Finfer S, et al: Global, regional, and national sepsis incidence and mortality, 1990-2017: Analysis for the Global Burden of Disease Study. Lancet. 395:200–211. 2020.PubMed/NCBI View Article : Google Scholar
3	Stearns-Kurosawa DJ, Osuchowski MF, Valentine C, Kurosawa S and Remick DG: The pathogenesis of sepsis. Annu Rev Pathol. 6:19–48. 2011.PubMed/NCBI View Article : Google Scholar
4	Yao YM, Luan YY, Zhang QH and Sheng ZY: Pathophysiological aspects of sepsis: An overview. Methods Mol Biol. 1237:5–15. 2015.PubMed/NCBI View Article : Google Scholar
5	He Q, Zhao L, Liu Y, Liu X, Zheng J, Yu H, Cai H, Ma J, Liu L, Wang P, et al: circ-SHKBP1 regulates the angiogenesis of U87 Glioma-exposed endothelial cells through miR-544a/FOXP1 and miR-379/FOXP2 pathways. Mol Ther Nucleic Acids. 10:331–348. 2018.PubMed/NCBI View Article : Google Scholar
6	Owens TW, Taylor RJ, Pahil KS, Bertani BR, Ruiz N, Kruse AC and Kahne D: Structural basis of unidirectional export of lipopolysaccharide to the cell surface. Nature. 567:550–553. 2019.PubMed/NCBI View Article : Google Scholar
7	Li Y, Orlando BJ and Liao M: Structural basis of lipopolysaccharide extraction by the LptB2FGC complex. Nature. 567:486–490. 2019.PubMed/NCBI View Article : Google Scholar
8	Beutler B and Rietschel ET: Innate immune sensing and its roots: The story of endotoxin. Nat Rev Immunol. 3:169–176. 2003.PubMed/NCBI View Article : Google Scholar
9	Wang Y, Zhang MX, Meng X, Liu FQ, Yu GS, Zhang C, Sun T, Wang XP, Li L, Wang YY, et al: Atorvastatin suppresses LPS-induced rapid upregulation of Toll-like receptor 4 and its signaling pathway in endothelial cells. Am J Physiol Heart Circ Physiol. 300:H1743–H1752. 2011.PubMed/NCBI View Article : Google Scholar
10	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-kappaB1 in human umbilical vein endothelial cells. Biochem Biophys Res Commun. 500:828–837. 2018.PubMed/NCBI View Article : Google Scholar
11	Wang Y, Li T, Wu B, Liu H, Luo J, Feng D and Shi Y: STAT1 regulates MD-2 expression in monocytes of sepsis via miR-30a. Inflammation. 37:1903–1911. 2014.PubMed/NCBI View Article : Google Scholar
12	Lv X, Zhang Y, Cui Y, Ren Y, Li R and Rong Q: Inhibition of microRNA155 relieves sepsisinduced liver injury through inactivating the JAK/STAT pathway. Mol Med Rep. 12:6013–6018. 2015.PubMed/NCBI View Article : Google Scholar
13	Seven M, Karatas OF, Duz MB and Ozen M: The role of miRNAs in cancer: From pathogenesis to therapeutic implications. Future Oncol. 10:1027–1048. 2014.PubMed/NCBI View Article : Google Scholar
14	van Rooij E and Olson EN: MicroRNA therapeutics for cardiovascular disease: Opportunities and obstacles. Nat Rev Drug Discov. 11:860–872. 2012.PubMed/NCBI View Article : Google Scholar
15	Liu YP, Gruber J, Haasnoot J, Konstantinova P and Berkhout B: RNAi-mediated inhibition of HIV-1 by targeting partially complementary viral sequences. Nucleic Acids Res. 37:6194–6204. 2009.PubMed/NCBI View Article : Google Scholar
16	Li M, Chen H, Chen L, Chen Y, Liu X and Mo D: miR-709 modulates LPS-induced inflammatory response through targeting GSK-3β. Int Immunopharmacol. 36:333–338. 2016.PubMed/NCBI View Article : Google Scholar
17	Alipoor SD, Mortaz E, Tabarsi P, Marjani M, Varahram M, Folkerts G, Garssen J and Adcock IM: miR-1224 expression is increased in human macrophages after infection with bacillus Calmette-Guerin (BCG). Iran J Allergy Asthma Immunol. 17:250–257. 2018.PubMed/NCBI
18	Guo X, Zhang Z, Zeng T, Lim YC, Wang Y, Xie X, Yang S, Huang C, Xu M, Tao L, et al: cAMP-MicroRNA-203-IFNγ network regulates subcutaneous white fat browning and glucose tolerance. Mol Metab. 28:36–47. 2019.PubMed/NCBI View Article : Google Scholar
19	Murphy AJ, Guyre PM and Pioli PA: Estradiol suppresses NF-kappa B activation through coordinated regulation of let-7a and miR-125b in primary human macrophages. J Immunol. 184:5029–5037. 2010.PubMed/NCBI View Article : Google Scholar
20	Xu G, Zhang Z, Xing Y, Wei J, Ge Z, Liu X, Zhang Y and Huang X: MicroRNA-149 negatively regulates TLR-triggered inflammatory response in macrophages by targeting MyD88. J Cell Biochem. 115:919–927. 2014.PubMed/NCBI View Article : Google Scholar
21	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
22	Xiao C, Calado DP, Galler G, Thai TH, Patterson HC, Wang J, Rajewsky N, Bender TP and Rajewsky K: miR-150 controls B cell differentiation by targeting the transcription factor c-Myb. Cell. 131:146–159. 2007.PubMed/NCBI View Article : Google Scholar
23	Luo JF, Xu J and Zheng JZ: Long non-coding RNA TTN-AS1 promotes cell proliferation and inhibits cell apoptosis in prostatic cancer by sponging miR-193a-5p. Eur Rev Med Pharmacol Sci. 23:7816–7825. 2019.PubMed/NCBI View Article : Google Scholar
24	Tsimikas S, Duff GW, Berger PB, Rogus J, Huttner K, Clopton P, Brilakis E, Kornman KS and Witztum JL: Pro-inflammatory interleukin-1 genotypes potentiate the risk of coronary artery disease and cardiovascular events mediated by oxidized phospholipids and lipoprotein(a). J Am Coll Cardiol. 63:1724–1734. 2014.PubMed/NCBI View Article : Google Scholar
25	Chaves de Souza JA, Nogueira AV, Chaves de Souza PP, Kim YJ, Silva Lobo C, Pimentel Lopes de Oliveira GJ, Cirelli JA, Garlet GP and Rossa C Jr: SOCS3 expression correlates with severity of inflammation, expression of proinflammatory cytokines, and activation of STAT3 and p38 MAPK in LPS-induced inflammation in vivo. Mediators Inflamm. 2013(650812)2013.PubMed/NCBI View Article : Google Scholar
26	Sheng B, Zhao L, Zang X, Zhen J and Chen W: miR-375 ameliorates sepsis by downregulating miR-21 level via inhibiting JAK2-STAT3 signaling. Biomed Pharmacother. 86:254–261. 2017.PubMed/NCBI View Article : Google Scholar
27	Liu F, Li Y, Jiang R, Nie C, Zeng Z, Zhao N, Huang C, Shao Q, Ding C, Qing C, et al: miR-132 inhibits lipopolysaccharide-induced inflammation in alveolar macrophages by the cholinergic anti-inflammatory pathway. Exp Lung Res. 41:261–269. 2015.PubMed/NCBI View Article : Google Scholar
28	Carlesso N, Frank DA and Griffin JD: Tyrosyl phosphorylation and DNA binding activity of signal transducers and activators of transcription (STAT) proteins in hematopoietic cell lines transformed by Bcr/Abl. J Exp Med. 183:811–820. 1996.PubMed/NCBI View Article : Google Scholar
29	Wex T, Grungreiff K, Schutte K, Stengritt M and Reinhold D: Expression analysis of zinc transporters in resting and stimulated human peripheral blood mononuclear cells. Biomed Rep. 2:217–222. 2014.PubMed/NCBI View Article : Google Scholar
30	Marengo E, Robotti E, Cecconi D, Hamdan M, Scarpa A and Righetti PG: Identification of the regulatory proteins in human pancreatic cancers treated with Trichostatin A by 2D-PAGE maps and multivariate statistical analysis. Anal Bioanal Chem. 379:992–1003. 2004.PubMed/NCBI View Article : Google Scholar
31	Gao B, Wang H, Lafdil F and Feng D: STAT proteins-key regulators of anti-viral responses, inflammation, and tumorigenesis in the liver. J Hepatol. 57:430–441. 2012.PubMed/NCBI View Article : Google Scholar
32	Seidel HM, Lamb P and Rosen J: Pharmaceutical intervention in the JAK/STAT signaling pathway. Oncogene. 19:2645–2656. 2000.PubMed/NCBI View Article : Google Scholar
33	Li X, Leung S, Qureshi S, Darnell JE Jr and Stark GR: Formation of STAT1-STAT2 heterodimers and their role in the activation of IRF-1 gene transcription by interferon-alpha. J Biol Chem. 271:5790–5794. 1996.PubMed/NCBI View Article : Google Scholar
34	Li Y, Yu C, Zhu WM, Xie Y, Qi X, Li N and Li JS: Triptolide ameliorates IL-10-deficient mice colitis by mechanisms involving suppression of IL-6/STAT3 signaling pathway and down-regulation of IL-17. Mol Immunol. 47:2467–2474. 2010.PubMed/NCBI View Article : Google Scholar
35	Chitnis T, Najafian N, Benou C, Salama AD, Grusby MJ, Sayegh MH and Khoury SJ: Effect of targeted disruption of STAT4 and STAT6 on the induction of experimental autoimmune encephalomyelitis. J Clin Invest. 108:739–747. 2001.PubMed/NCBI View Article : Google Scholar
36	Hranjec T and Sawyer RG: Management of infections in critically ill patients. Surg Infect (Larchmt). 15:474–478. 2014.PubMed/NCBI View Article : Google Scholar
37	Weledji EP and Ngowe MN: The challenge of intra-abdominal sepsis. Int J Surg. 11:290–295. 2013.PubMed/NCBI View Article : Google Scholar
38	Pearse RM, Moreno RP, Bauer P, Pelosi P, Metnitz P, Spies C, Vallet B, Vincent JL, Hoeft A and Rhodes A: European Surgical Outcomes Study (EuSOS) group for the Trials groups of the European Society of Intensive Care Medicine and the European Society of Anaesthesiology. Mortality after surgery in Europe: A 7 day cohort study. Lancet. 380:1059–1065. 2012.PubMed/NCBI View Article : Google Scholar
39	Song GY, Chung CS, Chaudry IH and Ayala A: IL-4-induced activation of the Stat6 pathway contributes to the suppression of cell-mediated immunity and death in sepsis. Surgery. 128:133–138. 2000.PubMed/NCBI View Article : Google Scholar
40	Spiegel JC, Lorenzen JM and Thum T: Role of microRNAs in immunity and organ transplantation. Expert Rev Mol Med. 13(e37)2011.PubMed/NCBI View Article : Google Scholar
41	Wu M, Gu JT, Yi B, Tang ZZ and Tao GC: microRNA-23b regulates the expression of inflammatory factors in vascular endothelial cells during sepsis. Exp Ther Med. 9:1125–1132. 2015.PubMed/NCBI View Article : Google Scholar
42	Gao N and Dong L: MicroRNA-146 regulates the inflammatory cytokines expression in vascular endothelial cells during sepsis. Pharmazie. 72:700–704. 2017.PubMed/NCBI View Article : Google Scholar
43	Wang Z, Ruan Z, Mao Y, Dong W, Zhang Y, Yin N and Jiang L: miR-27a is up regulated and promotes inflammatory response in sepsis. Cell Immunol. 290:190–195. 2014.PubMed/NCBI View Article : Google Scholar
44	Wang X, Wang X, Liu X, Wang X, Xu J, Hou S, Zhang X and Ding Y: miR-15a/16 are upreuglated in the serum of neonatal sepsis patients and inhibit the LPS-induced inflammatory pathway. Int J Clin Exp Med. 8:5683–5690. 2015.PubMed/NCBI
45	Estrella S, Garcia-Diaz DF, Codner E, Camacho-Guillen P and Perez-Bravo F: Expression of miR-22 and miR-150 in type 1 diabetes mellitus: Possible relationship with autoimmunity and clinical characteristics. Med Clin (Barc). 147:245–247. 2016.PubMed/NCBI View Article : Google Scholar : (In Spanish).
46	Hirschberger S, Hubner M, Strauss G, Effinger D, Bauer M, Weis S, Giamarellos-Bourboulis EJ and Kreth S: Identification of suitable controls for miRNA quantification in T-cells and whole blood cells in sepsis. Sci Rep. 9(15735)2019.PubMed/NCBI View Article : Google Scholar