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
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Research Support, Non-U.S. Gov't
MeSH terms
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Blotting, Western
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Butadienes / pharmacology
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Cells, Cultured
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Colon / cytology
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Enzyme-Linked Immunosorbent Assay
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Extracellular Signal-Regulated MAP Kinases / antagonists & inhibitors
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Extracellular Signal-Regulated MAP Kinases / metabolism
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Flavonoids / pharmacology
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Gene Expression / drug effects*
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Humans
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Imidazoles / pharmacology
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Interleukin-1 / genetics*
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Interleukin-1 / metabolism
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Interleukin-1beta / pharmacology*
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Microscopy, Confocal
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Myofibroblasts / drug effects*
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Myofibroblasts / metabolism
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Nitriles / pharmacology
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Pyridines / pharmacology
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RNA Interference
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Reverse Transcriptase Polymerase Chain Reaction
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Transcription Factor AP-1 / genetics
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Transcription Factor AP-1 / metabolism
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Transcription Factor RelA / genetics
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Transcription Factor RelA / metabolism
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Tumor Necrosis Factor-alpha / pharmacology
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p38 Mitogen-Activated Protein Kinases / antagonists & inhibitors
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p38 Mitogen-Activated Protein Kinases / metabolism
Substances
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Butadienes
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Flavonoids
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IL36G protein, human
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Imidazoles
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Interleukin-1
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Interleukin-1beta
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Nitriles
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Pyridines
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Transcription Factor AP-1
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Transcription Factor RelA
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Tumor Necrosis Factor-alpha
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U 0126
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Extracellular Signal-Regulated MAP Kinases
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p38 Mitogen-Activated Protein Kinases
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SB 203580
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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.