Helicobacter pylori lipopolysaccharide activates Rac1 and transcription of NADPH oxidase Nox1 and its organizer NOXO1 in guinea pig gastric mucosal cells

Tsukasa Kawahara, Motoyuki Kohjima, Yuki Kuwano, Hisano Mino, Shigetada Teshima-Kondo, Ryu Takeya, Shohko Tsunawaki, Akihiro Wada, Hideki Sumimoto, Kazuhito Rokutan

研究成果: ジャーナルへの寄稿記事

108 引用 (Scopus)

抄録

Primary cultures of guinea pig gastric mucosal cells express NADPH oxidase 1 (Nox1), a homolog of gp91phox, and produce superoxide anion (O 2-) at a rate of ∼100 nmol·mg protein -1· h-1 in response to Helicobacter pylori (H. pylori) lipopolysaccharide (LPS) from virulent type I strains. The upregulated O2- production also enhances H. pylori LPS-stimulated tumor necrosis factor-α or cyclooxygenase-2 mRNA expression, which suggests a potential role for Nox1 in the pathogenesis of H. pylori-associated diseases. The H. pylori LPS-stimulated O2- production in cultured gastric mucosal cells was inhibited by actinomycin D as well as cycloheximide, suggesting that the induction is regulated at the transcriptional level. The LPS treatment not only increased the Nox1 mRNA to a greater extent but also induced expression of the message-encoding, Nox-organizing protein 1 (NOXO1), a novel p47phox homolog required for Nox1 activity. In addition, H. pylori LPS activated Rac1; i.e., it converted Rac1 to the GTP-bound state. A phosphoinositide 3-kinase inhibitor, LY-294002, blocked H. pylori LPS-induced Rac1 activation and O2- generation without interfering with the expression of Nox1 and NOXO1 mRNA. O2 - production inhibited by LY-294002 was completely restored by transfection of an adenoviral vector encoding a constitutively active Rac1 but not an inactive Rac1 or a constitutively active Cdc42. These findings indicate that Rac1 plays a crucial role in Nox1 activation. Thus the H. pylori LPS-stimulated O2- production in gastric mucosal cells appears to require two distinct events: 1) transcriptional upregulation of Nox1 and NOXO1 and 2) activation of Rac1.

元の言語英語
ジャーナルAmerican Journal of Physiology - Cell Physiology
288
発行部数2 57-2
DOI
出版物ステータス出版済み - 2 1 2005

Fingerprint

NADPH Oxidase
Transcription
Stomach
Guinea Pigs
Proteins
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Chemical activation
Messenger RNA
1-Phosphatidylinositol 4-Kinase
Dactinomycin
Cyclooxygenase 2
Cycloheximide
Phosphatidylinositols
Guanosine Triphosphate
NADPH oxidase 1
Helicobacter pylori lipopolysaccharide
Helicobacter pylori
Superoxides
Transfection
Lipopolysaccharides

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

これを引用

Helicobacter pylori lipopolysaccharide activates Rac1 and transcription of NADPH oxidase Nox1 and its organizer NOXO1 in guinea pig gastric mucosal cells. / Kawahara, Tsukasa; Kohjima, Motoyuki; Kuwano, Yuki; Mino, Hisano; Teshima-Kondo, Shigetada; Takeya, Ryu; Tsunawaki, Shohko; Wada, Akihiro; Sumimoto, Hideki; Rokutan, Kazuhito.

:: American Journal of Physiology - Cell Physiology, 巻 288, 番号 2 57-2, 01.02.2005.

研究成果: ジャーナルへの寄稿記事

Kawahara, Tsukasa ; Kohjima, Motoyuki ; Kuwano, Yuki ; Mino, Hisano ; Teshima-Kondo, Shigetada ; Takeya, Ryu ; Tsunawaki, Shohko ; Wada, Akihiro ; Sumimoto, Hideki ; Rokutan, Kazuhito. / Helicobacter pylori lipopolysaccharide activates Rac1 and transcription of NADPH oxidase Nox1 and its organizer NOXO1 in guinea pig gastric mucosal cells. :: American Journal of Physiology - Cell Physiology. 2005 ; 巻 288, 番号 2 57-2.
@article{143aae06a0e6491990a3bc2c8b29b382,
title = "Helicobacter pylori lipopolysaccharide activates Rac1 and transcription of NADPH oxidase Nox1 and its organizer NOXO1 in guinea pig gastric mucosal cells",
abstract = "Primary cultures of guinea pig gastric mucosal cells express NADPH oxidase 1 (Nox1), a homolog of gp91phox, and produce superoxide anion (O 2-) at a rate of ∼100 nmol·mg protein -1· h-1 in response to Helicobacter pylori (H. pylori) lipopolysaccharide (LPS) from virulent type I strains. The upregulated O2- production also enhances H. pylori LPS-stimulated tumor necrosis factor-α or cyclooxygenase-2 mRNA expression, which suggests a potential role for Nox1 in the pathogenesis of H. pylori-associated diseases. The H. pylori LPS-stimulated O2- production in cultured gastric mucosal cells was inhibited by actinomycin D as well as cycloheximide, suggesting that the induction is regulated at the transcriptional level. The LPS treatment not only increased the Nox1 mRNA to a greater extent but also induced expression of the message-encoding, Nox-organizing protein 1 (NOXO1), a novel p47phox homolog required for Nox1 activity. In addition, H. pylori LPS activated Rac1; i.e., it converted Rac1 to the GTP-bound state. A phosphoinositide 3-kinase inhibitor, LY-294002, blocked H. pylori LPS-induced Rac1 activation and O2- generation without interfering with the expression of Nox1 and NOXO1 mRNA. O2 - production inhibited by LY-294002 was completely restored by transfection of an adenoviral vector encoding a constitutively active Rac1 but not an inactive Rac1 or a constitutively active Cdc42. These findings indicate that Rac1 plays a crucial role in Nox1 activation. Thus the H. pylori LPS-stimulated O2- production in gastric mucosal cells appears to require two distinct events: 1) transcriptional upregulation of Nox1 and NOXO1 and 2) activation of Rac1.",
author = "Tsukasa Kawahara and Motoyuki Kohjima and Yuki Kuwano and Hisano Mino and Shigetada Teshima-Kondo and Ryu Takeya and Shohko Tsunawaki and Akihiro Wada and Hideki Sumimoto and Kazuhito Rokutan",
year = "2005",
month = "2",
day = "1",
doi = "10.1152/ajpcell.00319.2004",
language = "English",
volume = "288",
journal = "American Journal of Physiology - Cell Physiology",
issn = "0363-6143",
publisher = "American Physiological Society",
number = "2 57-2",

}

TY - JOUR

T1 - Helicobacter pylori lipopolysaccharide activates Rac1 and transcription of NADPH oxidase Nox1 and its organizer NOXO1 in guinea pig gastric mucosal cells

AU - Kawahara, Tsukasa

AU - Kohjima, Motoyuki

AU - Kuwano, Yuki

AU - Mino, Hisano

AU - Teshima-Kondo, Shigetada

AU - Takeya, Ryu

AU - Tsunawaki, Shohko

AU - Wada, Akihiro

AU - Sumimoto, Hideki

AU - Rokutan, Kazuhito

PY - 2005/2/1

Y1 - 2005/2/1

N2 - Primary cultures of guinea pig gastric mucosal cells express NADPH oxidase 1 (Nox1), a homolog of gp91phox, and produce superoxide anion (O 2-) at a rate of ∼100 nmol·mg protein -1· h-1 in response to Helicobacter pylori (H. pylori) lipopolysaccharide (LPS) from virulent type I strains. The upregulated O2- production also enhances H. pylori LPS-stimulated tumor necrosis factor-α or cyclooxygenase-2 mRNA expression, which suggests a potential role for Nox1 in the pathogenesis of H. pylori-associated diseases. The H. pylori LPS-stimulated O2- production in cultured gastric mucosal cells was inhibited by actinomycin D as well as cycloheximide, suggesting that the induction is regulated at the transcriptional level. The LPS treatment not only increased the Nox1 mRNA to a greater extent but also induced expression of the message-encoding, Nox-organizing protein 1 (NOXO1), a novel p47phox homolog required for Nox1 activity. In addition, H. pylori LPS activated Rac1; i.e., it converted Rac1 to the GTP-bound state. A phosphoinositide 3-kinase inhibitor, LY-294002, blocked H. pylori LPS-induced Rac1 activation and O2- generation without interfering with the expression of Nox1 and NOXO1 mRNA. O2 - production inhibited by LY-294002 was completely restored by transfection of an adenoviral vector encoding a constitutively active Rac1 but not an inactive Rac1 or a constitutively active Cdc42. These findings indicate that Rac1 plays a crucial role in Nox1 activation. Thus the H. pylori LPS-stimulated O2- production in gastric mucosal cells appears to require two distinct events: 1) transcriptional upregulation of Nox1 and NOXO1 and 2) activation of Rac1.

AB - Primary cultures of guinea pig gastric mucosal cells express NADPH oxidase 1 (Nox1), a homolog of gp91phox, and produce superoxide anion (O 2-) at a rate of ∼100 nmol·mg protein -1· h-1 in response to Helicobacter pylori (H. pylori) lipopolysaccharide (LPS) from virulent type I strains. The upregulated O2- production also enhances H. pylori LPS-stimulated tumor necrosis factor-α or cyclooxygenase-2 mRNA expression, which suggests a potential role for Nox1 in the pathogenesis of H. pylori-associated diseases. The H. pylori LPS-stimulated O2- production in cultured gastric mucosal cells was inhibited by actinomycin D as well as cycloheximide, suggesting that the induction is regulated at the transcriptional level. The LPS treatment not only increased the Nox1 mRNA to a greater extent but also induced expression of the message-encoding, Nox-organizing protein 1 (NOXO1), a novel p47phox homolog required for Nox1 activity. In addition, H. pylori LPS activated Rac1; i.e., it converted Rac1 to the GTP-bound state. A phosphoinositide 3-kinase inhibitor, LY-294002, blocked H. pylori LPS-induced Rac1 activation and O2- generation without interfering with the expression of Nox1 and NOXO1 mRNA. O2 - production inhibited by LY-294002 was completely restored by transfection of an adenoviral vector encoding a constitutively active Rac1 but not an inactive Rac1 or a constitutively active Cdc42. These findings indicate that Rac1 plays a crucial role in Nox1 activation. Thus the H. pylori LPS-stimulated O2- production in gastric mucosal cells appears to require two distinct events: 1) transcriptional upregulation of Nox1 and NOXO1 and 2) activation of Rac1.

UR - http://www.scopus.com/inward/record.url?scp=19944427068&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=19944427068&partnerID=8YFLogxK

U2 - 10.1152/ajpcell.00319.2004

DO - 10.1152/ajpcell.00319.2004

M3 - Article

VL - 288

JO - American Journal of Physiology - Cell Physiology

JF - American Journal of Physiology - Cell Physiology

SN - 0363-6143

IS - 2 57-2

ER -