MiR-216a-5p alleviates LPS-induced inflammation in the human bronchial epithelial cell by inhibition of TGF-β1 signaling via down-regulating TGFBR2

Main Article Content

Shan Liu
Jianjun Li
Liya Hu

Keywords

bronchopneumonia, inflammation, miR-216a-5p, TGFBR2, TGFβ1 signaling

Abstract

Objective: Bronchopneumonia is a common respiratory infection disease and is the leading cause of hospitalization in children under 5 years of age. Inflammation is the primary response caused by bronchopneumonia. But the detailed underlying mechanism of inflammation in bronchopneumonia remains unclear. Therefore, this study focused on studying the effect of miR-216a-5p on inflammation induced by bronchopneumonia and investigate the potential mechanism underlying it.


Methods: Human bronchial epithelial cells (BEAS-2B) were stimulated using lipopolysaccha-rides (LPS) to trigger bronchopneumonia in vitro. The production of interleukin (IL)-1β, IL-6, and Tumor necrosis factor (TNF)-α was measured using the enzyme-linked immunosorbent assay. The luciferase assay was conducted to explore the relationship between miR-216a-5p and TGFBR2. Quantitative real-time polymerase chain reaction and western blot were used to detect the gene expression.


Results: miR-216a-5p gene expression decreased in BEAS-2B cells stimulated by LPS. Overexpression of miR-216a-5p suppressed the elevated levels of IL-1β, IL-6, and TNF-α induced by LPS. Transforming growth factor-beta receptor 2 (TGFBR2) proved to be a direct target of miR-216a-5p, and they negatively modulated TGFBR2 expression. In addition, overexpression of miR-216a-5p inhibited LPS-induced protein levels of TGFBR2, transforming growth factor (TGF)-β1, and phosphorylation of SMAD family member 2 (smad2),. This ectopic expression of miR-216a-5p was restored by overexpressed TGFBR2.


Conclusion: miR-216a-5p was decreased in LPS-stimulated BEAS-2B cells. Overexpressed miR-216a-5p suppressed LPS-induced inflammation in BEAS-2B cells by inhibition of TGF-β1 signal-ing via down-regulating TGFBR2. miR-216a-5p may be a valuable target for anti-inflammation treatment in bronchopneumonia.

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References

1. Lindström L, Tauni FA, Vargmar K. Bronchopneumonia in Swedish lambs: A study of pathological changes and bacterio-logical agents. Acta Veterinaria Scandinavica. 2018;60(1):1-8. 10.1186/s13028-018-0409-1

2. Gong M, Mao Z, Chen J, Ye X, Zhou X. microRNA-218 inhibits lipopolysaccharide-induced bronchial epithelial cell injury by targeting E2F2. J Biomater Tissue Eng. 2020;10(8):1161-1169. 10.1166/jbt.2020.2377

3. You C, Ran G, Wu X, Wang Y, Tian H, Fan J, et al. High immunoglobulin E level is associated with increased readmission in children with bronchopneumonia. Ther Adv Respir Dis. 2019; 13:1753466619879832. 10.1177/1753466619879832

4. Guan X, Silk BJ, Li W, Fleischauer AT, Xing X, Jiang X. Pneumonia incidence and mortality in Mainland China: Systematic review of Chinese and English literature, 1985-2008. PLoS One. 2010;5(7):e11721. 10.1371/journal.pone.0011721

5. Liu X, Meng J. Luteolin alleviates LPS-induced bronchopneumonia injury in vitro and in vivo by down-regulating microRNA-132 expression. Biomed Pharmacother. 2018;106:1641-1649. 10.1016/j.biopha.2018.07.094

6. Zhang Y, Zhu Y, Gao G, Zhou Z. Knockdown XIST alleviates LPS-induced WI-38 cell apoptosis and inflammation injury via targeting miR-370-3p/TLR4 in acute pneumonia. Cell Biochem Funct. 2019;37(5):348-58. 10.1002/cbf.3392

7. Vishnoi A, Rani S. MiRNA biogenesis and regulation of diseases: An overview. Methods Mol Bio. 2017;1509:1-10.10.1007/978-1-4939-6524-3_1

8. Reddy KB. MicroRNA (miRNA) in cancer. Cancer Cell Int. 2015;15(1):1-6. 10.1186/s12935-015-0185-1

9. Zhang L, Dong L, Tang Y, Li M, Zhang M. MiR-146b protects against the inflammation injury in pediatric pneumonia through MyD88/NF-κB signaling pathway. J Infect Dis. 2020;52(1):23-32. 10.1080/23744235.2019.1671987

10. Chi X, Ding B, Zhang L, Zhang J, Wang J, Zhang W. lncRNA GAS5 promotes M1 macrophage polarization via miR-455-5p/ SOCS3 pathway in childhood pneumonia. J Cell Physiol. 2019;234(8):13242-13251. 10.1002/jcp.27996

11. Chaoyang Y, Qingfeng B, Jinxing F. MiR-216a-5p protects 16HBE cells from H2O2-induced oxidative stress through targeting HMGB1/NF-kB pathway. Biochem Biophys Res Commun. 2019;508(2):416-420. 10.1016/j.bbrc.2018.11.060

12. Zhang J, Gao S, Zhang Y, Yi H, Xu M, Xu J, et al. MiR-216a-5p inhibits tumorigenesis in pancreatic cancer by targeting TPT1/mTORC1 and is mediated by LINC01133. Int J Biol Sci. 2020;16(14):2612. 10.7150/ijbs.46822

13. Kong F, Sun Y, Song W, Zhou Y, Zhu S. MiR-216a alleviates LPS-induced acute lung injury via regulating JAK2/STAT3 and NF-κB signaling. Hum Cell. 2020;33(1):67-78. 10.1007/s13577-019-00289-7

14. Peng R-R, Shang S-X, Zhao L-S, Long F-Q. MiR-216a-5p-containing exosomes suppress rTp17-induced inflammatory response by targeting TLR4. Biosci Rep. 2019;39(8): BSR20190686. 10.1042/BSR20190686

15. Sun Y, Hu B, Wang Y, Li Z, Wu J, Yang Y, et al. miR-216a-5p inhibits malignant progression in small cell lung cancer: Involvement of the Bcl-2 family proteins. Cancer Manag Res. 2018;10:4735. 10.2147/CMAR.S178380

16. Le Jan S, Muller C, Plee J, Durlach A, Bernard P, Antonicelli F. IL-23/IL-17 axis activates IL-1β-associated inflammasome in macrophages and generates an auto-inflammatory response in a subgroup of patients with bullous pemphigoid. Front Immunol. 2019;10:1972. 10.3389/fimmu.2019.01972

17. Yu T, Xu Y-Y, Zhang Y-Y, Li K-Y, Shao Y, Liu G. Plumbagin suppresses the human large cell lung cancer cell lines by inhibiting IL-6/STAT3 signaling in vitro. Int Immunopharmacol. 2018;55:290-296. 10.1016/j.intimp.2017.12.021

18. Han R, Zhang F, Wan C, Liu L, Zhong Q, Ding W. Effect of perfluorooctane sulphonate-induced Kupffer cell activation on hepatocyte proliferation through the NF-κB/TNF-α/ IL-6-dependent pathway. Chemosphere. 2018;200:283-294. 10.1016/j.chemosphere.2018.02.137

19. Fan Y, Gao Y, Rao J, Zhang F, Wang K, Zhang C. YAP-1 promotes tregs differentiation in hepatocellular carcinoma by enhancing TGFBR2 transcription. Cell Physiol Biochem. 2017;41(3):1189-1198. 10.1159/000464380

20. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25(4):402-408. 10.1006/meth.2001.1262

21. Sureshbabu A, Syed MA, Boddupalli CS, Dhodapkar MV, Homer RJ, Minoo P, et al. Conditional overexpression of TGFβ1 promotes pulmonary inflammation, apoptosis and mortality via TGFβR2 in the developing mouse lung. Respir Res. 2015;16(1):1-12. 10.1186/s12931-014-0162-6

22. Ganeshan K, Johnston LK, Bryce PJ. TGF-β1 limits the onset of innate lung inflammation by promoting mast cell-derived IL-6. J Immunol. 2013;190(11):5731-5738. 10.4049/jimmunol.1203362

23. Lykhmus O, Mishra N, Koval L, Kalashnyk O, Gergalova G, Uspenska K, et al. Molecular mechanisms regulating LPS-induced inflammation in the brain. Front Mol Neurosci. 2016;9:19. 10.3389/fnmol.2016.00019

24. Wang W, Li R. MiR-216a-5p alleviates chronic constriction injury-induced neuropathic pain in rats by targeting KDM3A and inactivating Wnt/β-catenin signaling pathway. Neurosci Res. 2020; S0168-0102(20)30421-1. 10.1016/j.neures.2020.08.001

25. Pu M, Chen J, Tao Z, Miao L, Qi X, Wang Y, et al. Regulatory network of miRNA on its target: Coordination between transcriptional and post-transcriptional regulation of gene expression. Cell Mol Life Sci. 2019;76(3):441-451. 10.1007/s00018-018-2940-7

26. Hart PJ, Deep S, Taylor AB, Shu Z, Hinck CS, Hinck AP. Crystal structure of the human TbetaR2 ectodomain--TGF-beta3 complex. Nat Struc Biol. 2002;9(3):203-208. 10.1038/nsb766

27. Massagué J, Seoane J, Wotton D. Smad transcription factors. Genes Dev. 2005;19(23):2783-810. 10.1101/gad.1350705

28. Katoh S, Matsumoto N, Tanaka H, Yasokawa N, Kittaka M, Kurose K, et al. Elevated levels of periostin and TGF-β1 in the bronchoalveolar lavage fluid of patients with idiopathic eosinophilic pneumonia. Asian Pac J Allergy Immunol.. 2020;38(3):208-213. 10.12932/AP-111018-0414