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Am J Physiol Gastrointest Liver Physiol 283: G986-G995, 2002. First published June 20, 2002; doi:10.1152/ajpgi.00482.2001
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Vol. 283, Issue 4, G986-G995, October 2002

Ammonia-induced apoptosis is accelerated at higher pH in gastric surface mucous cells

Hideo Suzuki1, Akinori Yanaka1, Takeshi Shibahara1, Hirofumi Matsui1, Akira Nakahara1, Naomi Tanaka1, Hiroshi Muto2, Takashi Momoi3, and Yasuo Uchiyama4

1 Departments of Gastroenterology and Endoscopy, Institute of Clinical Medicine, University of Tsukuba, Ibaraki 305-8575; 2 Toride Ishikai Hospital, Toride 300-0032; 3 Department of Biochemistry, Juntendo University School of Medicine, Tokyo 113-8421; and 4 Department of Cell Biology and Neuroscience, Osaka University Graduate School of Medicine, Osaka 565-0871, Japan

Gastric luminal ammonia produced by Helicobacter pylori has been shown to cause gastric mucosal injury. This study was conducted to examine the mechanisms by which gastric luminal ammonia causes apoptosis of gastric epithelial cells. Monolayers of GSM06 cells, developed from murine gastric surface mucous cells, were cultured in the absence or presence of 10-30 mM NH4Cl at ambient pH of 5.0, 6.0, and 7.0. In the presence of luminal NH4Cl, GSM06 cells showed 1) cell shrinkage and nuclear chromatin condensation, 2) DNA fragmentation into oligonucleosomes, 3) leakage of cytochrome c into cytosolic fraction without affecting bax expression, and 4) increases in activity of caspases-3 and -9. These changes were accentuated when the cells were cultured at pH 7.0. In the absence of NH4Cl, none of these changes was detected at any pH examined. These results suggest that gastric luminal ammonia, at concentrations detected in H. pylori-infected subjects, induces apoptosis of gastric epithelial cells by release of cytochrome c from mitochondria, followed by activation of caspases-9 and -3, especially at higher ambient pH.

Helicobacter pylori; gastric epithelial cells


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