Hyperosmotic stress enhances interleukin-1β expression in Helicobacter pylori-infected murine gastric epithelial cells in vitro

Background and Aim: Gastric cancer is associated not only with Helicobacter pylori (H. pylori) infection, but also with the intake of a high salt diet. Interleukin-1 beta (IL-1 beta) is highly expressed in H. pylori-infected gastric mucosa. The aim of the present study was to determine if hyperosmotic stress induces IL-1 beta expression in gastric epithelial cells in vitro. Method: Murine gastric epithelial cells, GSM06, were cultured with or without H. pylori (Sydney strain-1) at different osmolarities in the range of 300-450 mOsM. Expressions of IL-1 beta mRNA and mature IL-1 beta protein were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and an IL-1 beta enzyme-linked immunosorbent assay (ELISA), respectively. IL-1 beta converting enzyme (ICE) activity was measured by an ICE colorimetric assay. Apoptosis was evaluated by a single stranded-DNA assay. Results: Addition of H. pylori at 300 mosM caused significant increases in IL-1 beta mRNA, IL-1 beta protein, ICE activity and apoptosis. Hyperosmotic stress alone also caused upregulation of IL-1 beta mRNA and IL-1 beta protein, enhanced ICE activity and accelerated apoptosis. Hyperosmotic stress accentuated the increases in IL-1 beta mRNA, IL-1 beta protein, ICE activity and apoptosis induced by H. pylori alone. Enhancement of IL-1 beta protein release induced by hyperosmotic stress was significantly attenuated by an ICE inhibitor, Z-YVAD-FMK. Conclusions: Hyperosmotic stress enhances the release of bioactive mature IL-1 beta protein in H. pylori-infected gastric epithelial cells, in part by upregulating IL-1 beta mRNA expression, and in part by enhancing ICE activity. These results may explain the mechanisms by which chronic intake of a high salt diet increases the risk of gastric cancer among H. pylori-infected human subjects. (C) 2005 Blackwell Publishing Asia Pty Ltd.