Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

MiRNA-181a regulates Toll-like receptor agonist-induced inflammatory response in human fibroblasts

Genes & Immunity volume 15, pages 333–337 (2014)Cite this article

Subjects

Abstract

MicroRNAs (miRNAs) regulate the synthesis of cytokines in response to Toll-like receptor (TLR) activation. Our recent microarray study comparing normal and inflamed human dental pulps showed that miRNA-181 (miR-181) family is differentially expressed in the presence of inflammation. Prior studies have reported that the dental pulp, which is composed primarily of TLR4/2+ fibroblasts, expresses elevated levels of cytokines including interleukin-8 (IL-8) when inflamed. In this study, we employed an in-vitro model to determine the role of the miRNA-181 family in the TLR agonist-induced response in human fibroblasts. TLR4/2+ primary human dental pulp fibroblasts were stimulated with lipopolysaccharide from Porphyromonas gingivalis (Pg LPS), a known oral pathogen, and IL-8 and miR-181 expression measured. An inversely proportional relationship between IL-8 and miR-181a was observed. In-silico analysis identified a miR-181a-binding site on the 3′ untranslated region (UTR) of IL-8, which was confirmed by dual-luciferase assays. MiR-181a directly binds to the 3′UTR of IL-8, an important inflammatory component of the immune response, and modulates its levels. This is the very first report demonstrating miR-181a regulation of IL-8.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

References

  1. van Rooij E, Purcell AL, Levin AA . Developing microRNA therapeutics. Circ Res 2012; 110: 496–507.

    Article  CAS  PubMed  Google Scholar 

  2. Xie W, Li Z, Li M, Xu N, Zhang Y . miR-181a and inflammation: miRNA homeostasis response to inflammatory stimuli in vivo. Biochem Biophys Res Commun 2013; 430: 647–652.

    Article  CAS  PubMed  Google Scholar 

  3. Case SR, Martin RJ, Jiang D, Minor MN, Chu HW . MicroRNA-21 inhibits toll-like receptor 2 agonist-induced lung inflammation in mice. Exp Lung Res 2011; 37: 500–508.

    Article  CAS  PubMed  Google Scholar 

  4. Liu G, Friggeri A, Yang Y, Park YJ, Tsuruta Y, Abraham E . miR-147, a microRNA that is induced upon Toll-like receptor stimulation, regulates murine macrophage inflammatory responses. Proc Natl Acad Sci USA 2009; 106: 15819–15824.

    Article  CAS  PubMed  Google Scholar 

  5. Zhong S, Zhang S, Bair E, Nares S, Khan AA . Differential expression of microRNAs in normal and inflamed human pulps. J Endod 2012; 38: 746–752.

    Article  PubMed  Google Scholar 

  6. Love RM, Jenkinson HF . Invasion of dentinal tubules by oral bacteria. Crit Rev Oral Biol Med 2002; 13: 171–183.

    Article  CAS  PubMed  Google Scholar 

  7. Adachi T, Nakanishi T, Yumoto H, Hirao K, Takahashi K, Mukai K et al. Caries-related bacteria and cytokines induce CXCL10 in dental pulp. J Dental Res 2007; 86: 1217–1222.

    Article  CAS  Google Scholar 

  8. Huang GT, Potente AP, Kim JW, Chugal N, Zhang X . Increased interleukin-8 expression in inflamed human dental pulps. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 1999; 88: 214–220.

    Article  CAS  PubMed  Google Scholar 

  9. Kakehashi S, Stanley HR, Fitzgerald RJ . The effects of surgical exposures of dental pulps in germfree and conventional laboratory rats. J South Calif Dent Assoc 1966; 34: 449–451.

    CAS  PubMed  Google Scholar 

  10. Siqueira JF Jr., Rocas IN, Alves FR, Silva MG . Bacteria in the apical root canal of teeth with primary apical periodontitis. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2009; 107: 721–726.

    Article  PubMed  Google Scholar 

  11. Chan LT, Zhong S, Naqvi AR, Self-Fordham J, Nares S, Bair E et al. MicroRNAs: new insights into the pathogenesis of endodontic periapical disease. J Endod 2013; 39: 1498–1503.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Hirao K, Yumoto H, Takahashi K, Mukai K, Nakanishi T, Matsuo T . Roles of TLR2, TLR4, NOD2, and NOD1 in pulp fibroblasts. J Dent Res 2009; 88: 762–767.

    Article  CAS  PubMed  Google Scholar 

  13. Kawai T, Akira S . The role of pattern-recognition receptors in innate immunity: update on Toll-like receptors. Nat Immunol 2010; 11: 373–384.

    Article  CAS  PubMed  Google Scholar 

  14. Shimazu R, Akashi S, Ogata H, Nagai Y, Fukudome K, Miyake K et al. MD-2, a molecule that confers lipopolysaccharide responsiveness on Toll-like receptor 4. J Exp Med 1999; 189: 1777–1782.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Nagaoka S, Tokuda M, Sakuta T, Taketoshi Y, Tamura M, Takada H et al. Interleukin-8 gene expression by human dental pulp fibroblast in cultures stimulated with Prevotella intermedia lipopolysaccharide. J Endod 1996; 22: 9–12.

    Article  CAS  PubMed  Google Scholar 

  16. Park SH, Hsiao GY, Huang GT . Role of substance P and calcitonin gene-related peptide in the regulation of interleukin-8 and monocyte chemotactic protein-1 expression in human dental pulp. Int Endod J 2004; 37: 185–192.

    Article  CAS  PubMed  Google Scholar 

  17. Tokuda M, Sakuta T, Fushuku A, Torii M, Nagaoka S . Regulation of interleukin-6 expression in human dental pulp cell cultures stimulated with Prevotella intermedia lipopolysaccharide. J Endod 2001; 27: 273–277.

    Article  CAS  PubMed  Google Scholar 

  18. Sun X, Sit A, Feinberg MW . Role of miR-181 family in regulating vascular inflammation and immunity. Trends Cardiovasc Med 2013; 24: 105–112.

    Article  PubMed  PubMed Central  Google Scholar 

  19. Sun X, Icli B, Wara AK, Belkin N, He S, Kobzik L et al. MicroRNA-181b regulates NF-kappaB-mediated vascular inflammation. J Clin Invest 2012; 122: 1973–1990.

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Remus EW, Lyle AN, Weiss D, Landazuri N, Weber M, Searles C et al. miR181a protects against angiotensin II-induced osteopontin expression in vascular smooth muscle cells. Atherosclerosis 2013; 228: 168–174.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Harada A, Sekido N, Akahoshi T, Wada T, Mukaida N, Matsushima K . Essential involvement of interleukin-8 (IL-8) in acute inflammation. J Leukocyte Biol 1994; 56: 559–564.

    Article  CAS  PubMed  Google Scholar 

  22. Pollicino T, Bellinghieri L, Restuccia A, Raffa G, Musolino C, Alibrandi A et al. Hepatitis B virus (HBV) induces the expression of interleukin-8 that in turn reduces HBV sensitivity to interferon-alpha. Virology 2013; 444: 317–328.

    Article  CAS  PubMed  Google Scholar 

  23. Kaur M, Singh D . Neutrophil chemotaxis caused by chronic obstructive pulmonary disease alveolar macrophages: the role of CXCL8 and the receptors CXCR1/CXCR2. J Pharmacol Exp Ther 2013; 347: 173–180.

    Article  CAS  PubMed  Google Scholar 

  24. Noh MK, Jung M, Kim SH, Lee SR, Park KH, Kim DH et al. Assessment of IL-6, IL-8 and TNF-alpha levels in the gingival tissue of patients with periodontitis. Exp Ther Med 2013; 6:: 847–851.

    Article  Google Scholar 

  25. Karapanou V, Kempuraj D, Theoharides TC . Interleukin-8 is increased in gingival crevicular fluid from patients with acute pulpitis. J Endod 2008; 34: 148–151.

    Article  PubMed  Google Scholar 

  26. Hutchison ER, Kawamoto EM, Taub DD, Lal A, Abdelmohsen K, Zhang Y et al. Evidence for miR-181 involvement in neuroinflammatory responses of astrocytes. Glia 2013; 61: 1018–1028.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Xie W, Li M, Xu N, Lv Q, Huang N, He J et al. MiR-181a regulates inflammation responses in monocytes and macrophages. PLoS ONE 2013; 8: e58639.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Herath TD, Darveau RP, Seneviratne CJ, Wang CY, Wang Y, Jin L . Tetra- and penta-acylated lipid A structures of Porphyromonas gingivalis LPS differentially activate TLR4-mediated NF-kappaB signal transduction cascade and immuno-inflammatory response in human gingival fibroblasts. PLoS ONE 2013; 8: e58496.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Tabeta K, Yamazaki K, Akashi S, Miyake K, Kumada H, Umemoto T et al. Toll-like receptors confer responsiveness to lipopolysaccharide from Porphyromonas gingivalis in human gingival fibroblasts. Infect Immun 2000; 68: 3731–3735.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Hajishengallis G, Tapping RI, Harokopakis E, Nishiyama S, Ratti P, Schifferle RE et al. Differential interactions of fimbriae and lipopolysaccharide from Porphyromonas gingivalis with the Toll-like receptor 2-centred pattern recognition apparatus. Cell Microbiol 2006; 8: 1557–1570.

    Article  CAS  PubMed  Google Scholar 

  31. Liu X, Rennard S . MicroRNA and cytokines. Mol Cell Pharmacol 2011; 3: 143–151.

    CAS  Google Scholar 

  32. Henao-Mejia J, Williams A, Goff LA, Staron M, Licona-Limon P, Kaech SM et al. The MicroRNA miR-181 is a critical cellular metabolic rheostat essential for NKT cell ontogenesis and lymphocyte development and homeostasis. Immunity 2013; 38: 984–997.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Guo XK, Zhang Q, Gao L, Li N, Chen XX, Feng WH . Increasing expression of microRNA 181 inhibits porcine reproductive and respiratory syndrome virus replication and has implications for controlling virus infection. J Virol 2013; 87: 1159–1171.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. McDougall S, Dallon J, Sherratt J, Maini P . Fibroblast migration and collagen deposition during dermal wound healing: mathematical modelling and clinical implications. Philos Trans A Math Phys Eng Sci 2006; 364: 1385–1405.

    Article  CAS  PubMed  Google Scholar 

  35. Gura T . Chemokines take center stage in inflammatory ills. Science 1996; 272: 954–956.

    Article  CAS  PubMed  Google Scholar 

  36. Izumi T, Kobayashi I, Okamura K, Sakai H . Immunohistochemical study on the immunocompetent cells of the pulp in human non-carious and carious teeth. Arch Oral Biol 1995; 40: 609–614.

    Article  CAS  PubMed  Google Scholar 

  37. Olgart L, Brannstrom M, Johnson G . Invasion of bacteria into dentinal tubules. Experiments in vivo and in vitro. Acta odontologica Scandinavica 1974; 32: 61–70.

    Article  CAS  PubMed  Google Scholar 

  38. Hasselgren G, Calev D . Endodontics emergency treatment sound and simplified. N Y State Dent J 1994; 60: 31–33.

    CAS  PubMed  Google Scholar 

  39. Buckley M, Spangberg LS . The prevalence and technical quality of endodontic treatment in an American subpopulation. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 1995; 79: 92–100.

    Article  CAS  PubMed  Google Scholar 

  40. Pauley KM, Chan EK . MicroRNAs and their emerging roles in immunology. Ann NY Acad Sci 2008; 1143: 226–239.

    Article  CAS  PubMed  Google Scholar 

  41. Perry MM, Moschos SA, Williams AE, Shepherd NJ, Larner-Svensson HM, Lindsay MA . Rapid changes in microRNA-146a expression negatively regulate the IL-1beta-induced inflammatory response in human lung alveolar epithelial cells. J Immunol 2008; 180: 5689–5698.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Janssen HL, Reesink HW, Lawitz EJ, Zeuzem S, Rodriguez-Torres M, Patel K et al. Treatment of HCV infection by targeting microRNA. New Engl J Med 2013; 368: 1685–1694.

    Article  CAS  PubMed  Google Scholar 

  43. Ucar A, Gupta SK, Fiedler J, Erikci E, Kardasinski M, Batkai S et al. The miRNA-212/132 family regulates both cardiac hypertrophy and cardiomyocyte autophagy. Nat Commun 2012; 3: 1078.

    Article  PubMed  PubMed Central  Google Scholar 

  44. Yu Z, Willmarth NE, Zhou J, Katiyar S, Wang M, Liu Y et al. microRNA 17/20 inhibits cellular invasion and tumor metasis in breast cancer by herterotypic signalling. Proc Natl Acad Sci USA 2010; 107: 8231–8236.

    Article  CAS  PubMed  Google Scholar 

  45. Naqvi AR, Fordham JB, Khan A, Nares S . MicroRNAs responsive to Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis LPS modulate expression of genes regulating innate immunity in human macrophages. Innate Immun (e-pub ahead of print 23 September 2013; DOI:10.1177/1753425913501914).

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This research is supported by the National Institute for Dental and Craniofacial Research (NIDCR) R01DE021052, NIDCR T90DE021986 and the UNC School of Dentistry’s Next Generation of Oral Health Researchers (NextGen). Special thanks to Dr Richard Pestell (Jefferson Medical College, Philadelphia, PA, USA).

Author information

Authors and Affiliations

  1. Department of Endodontics, School of Dentistry, University of North Carolina, Chapel Hill, NC, USA

    J C Galicia & A A Khan

  2. Department of Periodontics, College of Dentistry, University of Illinois, Chicago, IL, USA

    A R Naqvi & S Nares

  3. Department of Orthodontics, School of Dentistry, University of North Carolina, Chapel Hill, NC, USA

    C-C Ko

  4. Regional Center for Neurosensory Disorders, School of Dentistry, University of North Carolina, Chapel Hill, NC, USA

    A A Khan

Authors
  1. J C Galicia
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A R Naqvi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C-C Ko
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S Nares
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A A Khan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to A A Khan.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Galicia, J., Naqvi, A., Ko, CC. et al. MiRNA-181a regulates Toll-like receptor agonist-induced inflammatory response in human fibroblasts. Genes Immun 15, 333–337 (2014). https://doi.org/10.1038/gene.2014.24

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/gene.2014.24

This article is cited by

Search

Advanced search

Quick links