Phosphatidylinositol 3-kinase/Akt signaling mediates interleukin-32α induction in human pancreatic periacinar myofibroblasts

Atsushi Nishida, Akira Andoh, Makoto Shioya, Shokei Kim-Mitsuyama, Atsushi Takayanagi, Yoshihide Fujiyama

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Interleukin (IL)-32 is a recently described proinflammatory cytokine, characterized by the induction of nuclear factor (NF)-κB activation. We studied IL-32α expression in human pancreatic periacinar myofibroblasts, which play important roles in the regulation of extracellular matrix metabolism and inflammatory responses in the pancreas. IL-32α protein expression was evaluated by Western blot analyses, and IL-32α mRNA expression was analyzed by Northern blot and real-time PCR analyses. IL-32α mRNA was weakly expressed without a stimulus, and its expression was markedly enhanced by IL-1β, IFN-γ, and TNF-α. IL-1β, IFN-γ, and TNF-α enhanced intracellular accumulation of IL-32α protein, but IL-32α was not detected in supernatants. Each cytokine dose and time dependently induced IL-32α mRNA expression. An inhibitor of phosphatidylinositol 3-kinase (LY294002) significantly suppressed IL-1β-, IFN-γ-, and TNF-α-induced IL-32α mRNA expression, although MAPK inhibitors had no effect. Akt activation in response to these cytokines was confirmed by Western blot. Furthermore, LY294002 suppressed both IL-1β- and TNF-α-induced NF-κB activation and IL-1β-, TNF-α-, and IFN-γ-induced activated protein-1 (AP-1) activation. Blockade of NF-κB and AP-1 activation by an adenovirus expressing a stable mutant form of IκBα and a dominant negative mutant of c-Jun markedly suppressed IL-1β-, IFN-γ-, and/or TNF-α-induced IL-32α mRNA expression. Human pancreatic periacinar myofibroblasts expressed IL-32α in response to IL-1β, TNF-α, and IFN-γ. IL-32α mRNA expression is dependent on interactions between the phosphatidylinositol 3-kinase/Akt-pathway and the NF-κB/AP-1 system.

Original languageEnglish
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume294
Issue number3
DOIs
Publication statusPublished - 2008 Mar

Fingerprint

Phosphatidylinositol 3-Kinase
Myofibroblasts
Interleukins
Interleukin-1
Messenger RNA
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Cytokines
Proteins
Western Blotting
Adenoviridae
Northern Blotting
Extracellular Matrix
Real-Time Polymerase Chain Reaction

Keywords

  • Cytokine
  • Inflammation
  • Pancreatitis

ASJC Scopus subject areas

  • Gastroenterology
  • Physiology

Cite this

Phosphatidylinositol 3-kinase/Akt signaling mediates interleukin-32α induction in human pancreatic periacinar myofibroblasts. / Nishida, Atsushi; Andoh, Akira; Shioya, Makoto; Kim-Mitsuyama, Shokei; Takayanagi, Atsushi; Fujiyama, Yoshihide.

In: American Journal of Physiology - Gastrointestinal and Liver Physiology, Vol. 294, No. 3, 03.2008.

Research output: Contribution to journalArticle

Nishida, Atsushi ; Andoh, Akira ; Shioya, Makoto ; Kim-Mitsuyama, Shokei ; Takayanagi, Atsushi ; Fujiyama, Yoshihide. / Phosphatidylinositol 3-kinase/Akt signaling mediates interleukin-32α induction in human pancreatic periacinar myofibroblasts. In: American Journal of Physiology - Gastrointestinal and Liver Physiology. 2008 ; Vol. 294, No. 3.
@article{5770ee2d0d5f4689bca56906fb390300,
title = "Phosphatidylinositol 3-kinase/Akt signaling mediates interleukin-32α induction in human pancreatic periacinar myofibroblasts",
abstract = "Interleukin (IL)-32 is a recently described proinflammatory cytokine, characterized by the induction of nuclear factor (NF)-κB activation. We studied IL-32α expression in human pancreatic periacinar myofibroblasts, which play important roles in the regulation of extracellular matrix metabolism and inflammatory responses in the pancreas. IL-32α protein expression was evaluated by Western blot analyses, and IL-32α mRNA expression was analyzed by Northern blot and real-time PCR analyses. IL-32α mRNA was weakly expressed without a stimulus, and its expression was markedly enhanced by IL-1β, IFN-γ, and TNF-α. IL-1β, IFN-γ, and TNF-α enhanced intracellular accumulation of IL-32α protein, but IL-32α was not detected in supernatants. Each cytokine dose and time dependently induced IL-32α mRNA expression. An inhibitor of phosphatidylinositol 3-kinase (LY294002) significantly suppressed IL-1β-, IFN-γ-, and TNF-α-induced IL-32α mRNA expression, although MAPK inhibitors had no effect. Akt activation in response to these cytokines was confirmed by Western blot. Furthermore, LY294002 suppressed both IL-1β- and TNF-α-induced NF-κB activation and IL-1β-, TNF-α-, and IFN-γ-induced activated protein-1 (AP-1) activation. Blockade of NF-κB and AP-1 activation by an adenovirus expressing a stable mutant form of IκBα and a dominant negative mutant of c-Jun markedly suppressed IL-1β-, IFN-γ-, and/or TNF-α-induced IL-32α mRNA expression. Human pancreatic periacinar myofibroblasts expressed IL-32α in response to IL-1β, TNF-α, and IFN-γ. IL-32α mRNA expression is dependent on interactions between the phosphatidylinositol 3-kinase/Akt-pathway and the NF-κB/AP-1 system.",
keywords = "Cytokine, Inflammation, Pancreatitis",
author = "Atsushi Nishida and Akira Andoh and Makoto Shioya and Shokei Kim-Mitsuyama and Atsushi Takayanagi and Yoshihide Fujiyama",
year = "2008",
month = "3",
doi = "10.1152/ajpgi.00535.2007",
language = "English",
volume = "294",
journal = "American Journal of Physiology - Heart and Circulatory Physiology",
issn = "0363-6135",
publisher = "American Physiological Society",
number = "3",

}

TY - JOUR

T1 - Phosphatidylinositol 3-kinase/Akt signaling mediates interleukin-32α induction in human pancreatic periacinar myofibroblasts

AU - Nishida, Atsushi

AU - Andoh, Akira

AU - Shioya, Makoto

AU - Kim-Mitsuyama, Shokei

AU - Takayanagi, Atsushi

AU - Fujiyama, Yoshihide

PY - 2008/3

Y1 - 2008/3

N2 - Interleukin (IL)-32 is a recently described proinflammatory cytokine, characterized by the induction of nuclear factor (NF)-κB activation. We studied IL-32α expression in human pancreatic periacinar myofibroblasts, which play important roles in the regulation of extracellular matrix metabolism and inflammatory responses in the pancreas. IL-32α protein expression was evaluated by Western blot analyses, and IL-32α mRNA expression was analyzed by Northern blot and real-time PCR analyses. IL-32α mRNA was weakly expressed without a stimulus, and its expression was markedly enhanced by IL-1β, IFN-γ, and TNF-α. IL-1β, IFN-γ, and TNF-α enhanced intracellular accumulation of IL-32α protein, but IL-32α was not detected in supernatants. Each cytokine dose and time dependently induced IL-32α mRNA expression. An inhibitor of phosphatidylinositol 3-kinase (LY294002) significantly suppressed IL-1β-, IFN-γ-, and TNF-α-induced IL-32α mRNA expression, although MAPK inhibitors had no effect. Akt activation in response to these cytokines was confirmed by Western blot. Furthermore, LY294002 suppressed both IL-1β- and TNF-α-induced NF-κB activation and IL-1β-, TNF-α-, and IFN-γ-induced activated protein-1 (AP-1) activation. Blockade of NF-κB and AP-1 activation by an adenovirus expressing a stable mutant form of IκBα and a dominant negative mutant of c-Jun markedly suppressed IL-1β-, IFN-γ-, and/or TNF-α-induced IL-32α mRNA expression. Human pancreatic periacinar myofibroblasts expressed IL-32α in response to IL-1β, TNF-α, and IFN-γ. IL-32α mRNA expression is dependent on interactions between the phosphatidylinositol 3-kinase/Akt-pathway and the NF-κB/AP-1 system.

AB - Interleukin (IL)-32 is a recently described proinflammatory cytokine, characterized by the induction of nuclear factor (NF)-κB activation. We studied IL-32α expression in human pancreatic periacinar myofibroblasts, which play important roles in the regulation of extracellular matrix metabolism and inflammatory responses in the pancreas. IL-32α protein expression was evaluated by Western blot analyses, and IL-32α mRNA expression was analyzed by Northern blot and real-time PCR analyses. IL-32α mRNA was weakly expressed without a stimulus, and its expression was markedly enhanced by IL-1β, IFN-γ, and TNF-α. IL-1β, IFN-γ, and TNF-α enhanced intracellular accumulation of IL-32α protein, but IL-32α was not detected in supernatants. Each cytokine dose and time dependently induced IL-32α mRNA expression. An inhibitor of phosphatidylinositol 3-kinase (LY294002) significantly suppressed IL-1β-, IFN-γ-, and TNF-α-induced IL-32α mRNA expression, although MAPK inhibitors had no effect. Akt activation in response to these cytokines was confirmed by Western blot. Furthermore, LY294002 suppressed both IL-1β- and TNF-α-induced NF-κB activation and IL-1β-, TNF-α-, and IFN-γ-induced activated protein-1 (AP-1) activation. Blockade of NF-κB and AP-1 activation by an adenovirus expressing a stable mutant form of IκBα and a dominant negative mutant of c-Jun markedly suppressed IL-1β-, IFN-γ-, and/or TNF-α-induced IL-32α mRNA expression. Human pancreatic periacinar myofibroblasts expressed IL-32α in response to IL-1β, TNF-α, and IFN-γ. IL-32α mRNA expression is dependent on interactions between the phosphatidylinositol 3-kinase/Akt-pathway and the NF-κB/AP-1 system.

KW - Cytokine

KW - Inflammation

KW - Pancreatitis

UR - http://www.scopus.com/inward/record.url?scp=41549109283&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=41549109283&partnerID=8YFLogxK

U2 - 10.1152/ajpgi.00535.2007

DO - 10.1152/ajpgi.00535.2007

M3 - Article

C2 - 18239058

AN - SCOPUS:41549109283

VL - 294

JO - American Journal of Physiology - Heart and Circulatory Physiology

JF - American Journal of Physiology - Heart and Circulatory Physiology

SN - 0363-6135

IS - 3

ER -