Helicobacter pylori-associated ammonia production enhances neutrophil- dependent gastric mucosal cell injury

M. Suzuki, S. Miura, M. Suematsu, D. Fukumura, I. Kurose, Hidekazu Suzuki, A. Kai, Y. Kudoh, M. Ohashi, M. Tsuchiya

Research output: Contribution to journalArticle

192 Citations (Scopus)

Abstract

The role of neutrophil and its chlorinated oxidant were investigated in Helicobacter pylori-induced gastric mucosal injury in vitro. Luminol- dependent chemiluminescence (Ch(L)) was used to detect neutrophil-derived oxidants. Ch(L) 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.

Original languageEnglish
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume263
Issue number5 26-5
Publication statusPublished - 1992

Fingerprint

Ammonia
Helicobacter pylori
Stomach
Neutrophils
Wounds and Injuries
Oxidants
Hypochlorous Acid
Luminescence
Intracellular Space
Luminol
Respiratory Burst
Urease
Taurine
Fluorescein
Methionine
Catalase
Hydrogen Peroxide
Urea
Cultured Cells
Fluorescence

Keywords

  • monochloramine

ASJC Scopus subject areas

  • Physiology
  • Gastroenterology

Cite this

Helicobacter pylori-associated ammonia production enhances neutrophil- dependent gastric mucosal cell injury. / Suzuki, M.; Miura, S.; Suematsu, M.; Fukumura, D.; Kurose, I.; Suzuki, Hidekazu; Kai, A.; Kudoh, Y.; Ohashi, M.; Tsuchiya, M.

In: American Journal of Physiology - Gastrointestinal and Liver Physiology, Vol. 263, No. 5 26-5, 1992.

Research output: Contribution to journalArticle

Suzuki, M, Miura, S, Suematsu, M, Fukumura, D, Kurose, I, Suzuki, H, Kai, A, Kudoh, Y, Ohashi, M & Tsuchiya, M 1992, 'Helicobacter pylori-associated ammonia production enhances neutrophil- dependent gastric mucosal cell injury', American Journal of Physiology - Gastrointestinal and Liver Physiology, vol. 263, no. 5 26-5.
Suzuki, M. ; Miura, S. ; Suematsu, M. ; Fukumura, D. ; Kurose, I. ; Suzuki, Hidekazu ; Kai, A. ; Kudoh, Y. ; Ohashi, M. ; Tsuchiya, M. / Helicobacter pylori-associated ammonia production enhances neutrophil- dependent gastric mucosal cell injury. In: American Journal of Physiology - Gastrointestinal and Liver Physiology. 1992 ; Vol. 263, No. 5 26-5.
@article{c4539a29996c43c19e55a8116db525a7,
title = "Helicobacter pylori-associated ammonia production enhances neutrophil- dependent gastric mucosal cell injury",
abstract = "The role of neutrophil and its chlorinated oxidant were investigated in Helicobacter pylori-induced gastric mucosal injury in vitro. Luminol- dependent chemiluminescence (Ch(L)) was used to detect neutrophil-derived oxidants. Ch(L) 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.",
keywords = "monochloramine",
author = "M. Suzuki and S. Miura and M. Suematsu and D. Fukumura and I. Kurose and Hidekazu Suzuki and A. Kai and Y. Kudoh and M. Ohashi and M. Tsuchiya",
year = "1992",
language = "English",
volume = "263",
journal = "American Journal of Physiology - Heart and Circulatory Physiology",
issn = "0363-6135",
publisher = "American Physiological Society",
number = "5 26-5",

}

TY - JOUR

T1 - Helicobacter pylori-associated ammonia production enhances neutrophil- dependent gastric mucosal cell injury

AU - Suzuki, M.

AU - Miura, S.

AU - Suematsu, M.

AU - Fukumura, D.

AU - Kurose, I.

AU - Suzuki, Hidekazu

AU - Kai, A.

AU - Kudoh, Y.

AU - Ohashi, M.

AU - Tsuchiya, M.

PY - 1992

Y1 - 1992

N2 - The role of neutrophil and its chlorinated oxidant were investigated in Helicobacter pylori-induced gastric mucosal injury in vitro. Luminol- dependent chemiluminescence (Ch(L)) was used to detect neutrophil-derived oxidants. Ch(L) 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.

AB - The role of neutrophil and its chlorinated oxidant were investigated in Helicobacter pylori-induced gastric mucosal injury in vitro. Luminol- dependent chemiluminescence (Ch(L)) was used to detect neutrophil-derived oxidants. Ch(L) 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.

KW - monochloramine

UR - http://www.scopus.com/inward/record.url?scp=0026492780&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026492780&partnerID=8YFLogxK

M3 - Article

VL - 263

JO - American Journal of Physiology - Heart and Circulatory Physiology

JF - American Journal of Physiology - Heart and Circulatory Physiology

SN - 0363-6135

IS - 5 26-5

ER -