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Journal of Pharmacology and Experimental Therapeutics

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Research ArticleGASTROINTESTINAL, HEPATIC, PULMONARY, AND RENAL

Tumor Necrosis Factor-α-Induced Cytokine-Induced Neutrophil Chemoattractant-1 (CINC-1) Production by Rat Gastric Epithelial Cells: Role of Reactive Oxygen Species and Nuclear Factor-κB

Osamu Handa, Yuji Naito, Tomohisa Takagi, Makoto Shimozawa, Satoshi Kokura, Norimasa Yoshida, Hirofumi Matsui, Gediminas Cepinskas, Peter R. Kvietys and Toshikazu Yoshikawa
Journal of Pharmacology and Experimental Therapeutics May 2004, 309 (2) 670-676; DOI: https://doi.org/10.1124/jpet.103.062216
Osamu Handa
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Yuji Naito
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Tomohisa Takagi
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Makoto Shimozawa
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Satoshi Kokura
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Norimasa Yoshida
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Hirofumi Matsui
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Gediminas Cepinskas
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Peter R. Kvietys
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Toshikazu Yoshikawa
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Abstract

Rat cytokine-induced neutrophil chemoattractant-1 (CINC-1), a counterpart of the human growth-regulated oncogene product (GRO), has been suggested to participate in neutrophil recruitment in an experimental model of gastritis in rat. However, the mechanism(s) involved in regulation of CINC-1 production by the gastric mucosa remains unclear. The aim of this study was to investigate the mechanism(s) of CINC-1 production by rat gastric mucosa in vitro. All experiments were performed using rat normal gastric mucosal cell line (RGM-1). RGM-1s were stimulated with tumor necrosis factor (TNF)-α, and CINC-1 mRNA levels (reverse transcription-polymerase chain reaction) and protein secretion (enzyme-linked immunosorbent assay) were assessed. The production of reactive oxygen species (ROS) and nuclear factor (NF)-κB activation (translocation to the nuclei) in response to TNF-α stimulation was evaluated using fluorescence microscopy in the presence or absence of the inhibitors of mitochondrial electron flow and NF-κB activation. Stimulation of RGM-1 cells with TNF-α resulted in an increase in intracellular oxidative stress, NF-κB translocation to the nuclei, and up-regulation of CINC-1 mRNA and protein, which was prevented by interfering with mitochondria-dependent ROS production and NF-κB activation. Taken together, these findings indicate that CINC-1, a counterpart of the human GRO, production by rat gastric epithelial cells in response to TNF-α stimulation is an oxidant stress-mediated and NF-κB-dependent event.

Footnotes

  • This work was supported by Grant-in-Aid for Scientific Research 14570943 (to Y.N.) and 15390178 (to T.Y.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

  • Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.

  • DOI: 10.1124/jpet.103.062216.

  • ABBREVIATIONS. IL-8, interleukin-8; GRO, growth-regulated oncogene product; CINC-1, cytokine-induced neutrophil chemoattractant-1; TNF-α, tumor necrosis factor-α; IL-1β, interleukin-1β; LPS, lipopolysaccharide; RGM-1, rat gastric mucosal cells; PSI, proteasome inhibitor; NF-κB, nuclear factor-κB; PDTC, pyrrolidine dithiocarbamate; DPI, diphenyleneiodonium; ELISA, enzyme-linked immunosorbent assay; PBS, phosphate-buffered saline; RT, reverse transcription; PCR, polymerase chain reaction ROS, reactive oxygen species; CH2DCF-DA, 5-(and 6-)carboxy-2′,7′-dichlorodihydrofluorescein diacetate; DHR-123, dihydrorhodamine-123.

    • Received October 31, 2003.
    • Accepted January 23, 2004.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 309 (2)
Journal of Pharmacology and Experimental Therapeutics
Vol. 309, Issue 2
1 May 2004
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Research ArticleGASTROINTESTINAL, HEPATIC, PULMONARY, AND RENAL

Tumor Necrosis Factor-α-Induced Cytokine-Induced Neutrophil Chemoattractant-1 (CINC-1) Production by Rat Gastric Epithelial Cells: Role of Reactive Oxygen Species and Nuclear Factor-κB

Osamu Handa, Yuji Naito, Tomohisa Takagi, Makoto Shimozawa, Satoshi Kokura, Norimasa Yoshida, Hirofumi Matsui, Gediminas Cepinskas, Peter R. Kvietys and Toshikazu Yoshikawa
Journal of Pharmacology and Experimental Therapeutics May 1, 2004, 309 (2) 670-676; DOI: https://doi.org/10.1124/jpet.103.062216

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Research ArticleGASTROINTESTINAL, HEPATIC, PULMONARY, AND RENAL

Tumor Necrosis Factor-α-Induced Cytokine-Induced Neutrophil Chemoattractant-1 (CINC-1) Production by Rat Gastric Epithelial Cells: Role of Reactive Oxygen Species and Nuclear Factor-κB

Osamu Handa, Yuji Naito, Tomohisa Takagi, Makoto Shimozawa, Satoshi Kokura, Norimasa Yoshida, Hirofumi Matsui, Gediminas Cepinskas, Peter R. Kvietys and Toshikazu Yoshikawa
Journal of Pharmacology and Experimental Therapeutics May 1, 2004, 309 (2) 670-676; DOI: https://doi.org/10.1124/jpet.103.062216
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