Interleukin (IL)-1β Is a Strong Inducer of IL-36γ Expression in Human Colonic Myofibroblasts

PLoS One. 2015 Nov 12;10(11):e0138423. doi: 10.1371/journal.pone.0138423. eCollection 2015.

Abstract

Backgrounds and aims: Interleukin (IL)-36 cytokines are members of the IL-1 cytokine family. In this study, we investigated the expression of IL-36γ in human colonic myofibroblasts to explore the molecular mechanisms underlying IL-36γ induction.

Materials and methods: IL-36 mRNA was analyzed by real-time PCR method. Secretion of IL-36γ protein was evaluated by Western blot and ELISA analyses. Molecular mechanism of IL-36γ induction was evaluated by siRNA analyses and immunofluorescence experiments.

Results: IL-36γ mRNA expression was scarcely detected in the cells without stimulation. IL-1β induced a marked increase of IL-36γ mRNA expression. TNF-α markedly enhanced IL-1β-induced IL-36γ mRNA expression. These responses were confirmed at the protein levels. The inhibitors for ERK1/2 (PD98059 and U0216) and a p38 MAPK (SB203580) significantly reduced the IL-1β-induced IL-36γ mRNA expression. In addition, the siRNAs specific for NF-κB p65 and AP-1 (c-Jun) significantly reduced the expression of IL-1β-induced IL-36γ mRNA.

Conclusions: Colonic myofibroblasts are cellular source of IL-36γ in the intestine. IL-36γ expression was induced by the combination of IL-1β and TNF-α via activation of MAPKs and transcription factors, NF-κB and AP-1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Butadienes / pharmacology
  • Cells, Cultured
  • Colon / cytology
  • Enzyme-Linked Immunosorbent Assay
  • Extracellular Signal-Regulated MAP Kinases / antagonists & inhibitors
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Flavonoids / pharmacology
  • Gene Expression / drug effects*
  • Humans
  • Imidazoles / pharmacology
  • Interleukin-1 / genetics*
  • Interleukin-1 / metabolism
  • Interleukin-1beta / pharmacology*
  • Microscopy, Confocal
  • Myofibroblasts / drug effects*
  • Myofibroblasts / metabolism
  • Nitriles / pharmacology
  • Pyridines / pharmacology
  • RNA Interference
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcription Factor AP-1 / genetics
  • Transcription Factor AP-1 / metabolism
  • Transcription Factor RelA / genetics
  • Transcription Factor RelA / metabolism
  • Tumor Necrosis Factor-alpha / pharmacology
  • p38 Mitogen-Activated Protein Kinases / antagonists & inhibitors
  • p38 Mitogen-Activated Protein Kinases / metabolism

Substances

  • Butadienes
  • Flavonoids
  • IL36G protein, human
  • Imidazoles
  • Interleukin-1
  • Interleukin-1beta
  • Nitriles
  • Pyridines
  • Transcription Factor AP-1
  • Transcription Factor RelA
  • Tumor Necrosis Factor-alpha
  • U 0126
  • Extracellular Signal-Regulated MAP Kinases
  • p38 Mitogen-Activated Protein Kinases
  • SB 203580
  • 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one

Grants and funding

This study was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (15K08967), a grant for the Intractable Diseases from the Ministry of Health, Labor and Welfare of Japan (067), a grant from the Practical Research Project for Rare/Intractable Diseases from Japan Agency for Medical Research and development, AMED (The Japan Agency of Medical Research and Development) (15AeK0109047h0002), and a grant from Smoking Research Foundation (1848) (www.srf.or.jp/english/index.html). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.