Helicobacter pylori-associated ammonia production enhances neutrophil-dependent gastric mucosal cell injury.
The role of neutrophil and its chlorinated oxidant were investigated in Helicobacter pylori-induced gastric mucosal injury in vitro. Luminol-dependent chemiluminescence (ChL) was used to detect neutrophil-derived oxidants. ChL activity was significantly elevated when neutrophils were incubated in H. pylori, indicating that H. pylori actually elicits oxidative burst of neutrophils. To assess whether H. pylori-activated neutrophils exert the cytotoxicity for gastric mucosal cells, rabbit gastric mucosal cell was monolayered in culture wells and labeled with a fluorescence dye, 2',7'-bis(2-carboxyethyl)-5(6)carboxy-fluorescein, which is retained in the intracellular space as long as the cell membrane is intact. Labeled cells were coincubated with neutrophils and H. pylori. We inferred from the cytotoxicity index (specific %cytotoxicity), which was calculated from fluorometrical measurements of supernatant and lysate, that the mucosal cells were significantly damaged by H. pylori-activated neutrophils. This injury was largely attenuated by eliminating urea from the incubation mixture or by acetohydroxamic acid, a potent urease inhibitor. Additionally, the scavengers of neutrophil-derived oxidants, including taurine, methionine, and catalase, also attenuated this injury. Cultured mucosal cells that were exposed to the solution containing monochloramine (an oxidant yielded by reaction of hypochlorous acid and ammonia) were highly damaged compared with cells exposed to hypochlorous acid or hydrogen peroxide at physiological concentrations. These data suggest that H. pylori-activated neutrophils promote gastric mucosal cell injury and that monochloramine plays a unique and important role in this process.
Suzuki
M
M
Department of Internal Medicine, School of Medicine, Keio University, Tokyo, Japan.
Miura
S
S
Suematsu
M
M
Fukumura
D
D
Kurose
I
I
Suzuki
H
H
Kai
A
A
Kudoh
Y
Y
Ohashi
M
M
Tsuchiya
M
M
D000641:Ammonia / Q000378:metabolism*; D002470:Cell Survival; D005452:Fluoresceins; D005753:Gastric Mucosa / Q000187:drug effects / Q000473:pathology*; D016480:Helicobacter pylori / Q000502:physiology*; D006801:Humans; D008165:Luminol; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D009504:Neutrophils / Q000378:metabolism / Q000473:pathology / Q000502:physiology*; D016877:Oxidants / Q000378:metabolism / Q000494:pharmacology
D005452:Fluoresceins; D016877:Oxidants; D008165:Luminol; D000641:Ammonia; C043829:2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein