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GASTROINTESTINAL, HEPATIC, PULMONARY, AND RENAL

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

Vascular Cell Biology/Inflammation Program, Lawson Health Research Institute, London, Ontario, Canada (O.H., G.C., P.R.K.); Department of Inflammation and Immunology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kamigyo-ku, Kyoto, Japan (O.H., T.T., M.S. T.Y.); Department of Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kamigyo-ku, Kyoto, Japan (Y.N., S.K., N.Y.); and Division of Gastroenterology, Institute of Clinical Medicine, University of Tsukuba, Tsukuba, Ibaraki, Japan (H.M.)

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)-, 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)-B activation (translocation to the nuclei) in response to TNF- stimulation was evaluated using fluorescence microscopy in the presence or absence of the inhibitors of mitochondrial electron flow and NF-B activation. Stimulation of RGM-1 cells with TNF- resulted in an increase in intracellular oxidative stress, NF-B 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-B 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- stimulation is an oxidant stress-mediated and NF-B-dependent event.

Address correspondence to: Dr. Yuji Naito, Department of Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kajiicho, Kawaramachidori Hirokouji Agaru, Kamigyou-ku, Kyoto 602-8566, Japan. E-mail: ynaito{at}koto.kpu-m.ac.jp

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