Helicobacter pylori induces apoptosis of rat gastric parietal cells

被引:53
作者
Neu, B
Randlkofer, P
Neuhofer, M
Voland, P
Mayerhofer, A
Gerhard, M
Schepp, W
Prinz, C
机构
[1] Tech Univ Munich, Med Klin 2, D-81675 Munich, Germany
[2] Tech Univ Munich, Krankenhaus Munchen Bogenhausen, D-81675 Munich, Germany
[3] Univ Munich, Inst Anat, Munich, Germany
来源
AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY | 2002年 / 283卷 / 02期
关键词
nuclear factor-kappa B;
D O I
10.1152/ajpgi.00546.2001
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
Gastric Helicobacter pylori infection may lead to multifocal atrophic corpus gastritis associated with loss of epithelial cells as well as glandular structures. The current work investigated H. pylori effects on cell death of isolated, nontransformed rat parietal cells (PC). Highly enriched rat PC (>97%) were isolated from gastric mucosa and cultured in serum-free medium over 24 h. The cells were cocultured over 8 h with cytotoxin-associated immunodominant protein (cagA)(+)/vacuolating toxin (vacA)(+) or with cagA(-)/vacA(-) H. pylori laboratory strains and also with H. pylori mutants deleted in several genes of the cag pathogenicity island. Staphylococcus aureus or Campylobacter jejuni were used as controls. Apoptosis was determined by terminal deoxynucleotidyl transferase dUTP nick-end labeling staining and electron microscopy. Interleukin (IL)-8 and cytokine-induced neutrophil chemoattractant (CINC)-1 secretion was measured by ELISA. Activation of nuclear factor-kappaB (NF-kappaB) was studied in nuclear extracts of PC by electrophoretic mobility shift assay. Apoptosis of PC was induced in a concentration- and time-dependent manner by cagA(+)/vacA(+) H. pylori strains but not by cagA(-)/vacA(-) negative strains or by the cagE knockout mutant. S. aureus and C. jejuni had no effect. PC showed no IL-8 or CINC-1 secretion on exposure to cagA(+)/vacA(+) H. pylori. cagA(+)/vacA(+) strains induced activation of NF-kappaB complexes in nuclear extracts of PC, which were composed of p65 and p50 subunits. No significant stimulation of NF-kappaB activation was detected by incubation of PC with the cagE knockout mutant. Preincubation of PC with antisense but not missense oligodeoxynucleotides against the p65 subunit significantly reduced DNA binding to the kappaB recognition sequence. The p65 oligonucleotides as well as the proteasome inhibitor N-CBZ-isoleucin-glutamin-(o-t-butyl-)-alanin-leucin and the nitric oxide synthase inhibitor N-G-monomethyl-L-arginine completely prevented PC apoptosis induced by cagA(+)/vacA(+) strains. In summary, cagE presence appears to be essential for H. pylori-induced apoptosis of gastric parietal cells, and this effect is dependent on the activation of NF-kappaB and production of nitric oxide.
引用
收藏
页码:G309 / G318
页数:10
相关论文
共 36 条
[1]   Helicobacter pylori inhibits gastric cell cycle progression [J].
Ahmed, A ;
Smoot, D ;
Littleton, G ;
Tackey, R ;
Walters, CS ;
Kashanchi, F ;
Allen, CR ;
Ashktorab, H .
MICROBES AND INFECTION, 2000, 2 (10) :1159-1169
[2]   Mechanisms involved in Helicobacter pylori-induced interleukin-8 production by a gastric cancer cell line, MKN45 [J].
Aihara, M ;
Tsuchimoto, D ;
Takizawa, H ;
Azuma, A ;
Wakebe, H ;
Ohmoto, Y ;
Imagawa, K ;
Kikuchi, M ;
Mukaida, N ;
Matsushima, K .
INFECTION AND IMMUNITY, 1997, 65 (08) :3218-3224
[3]  
Ando T, 1996, AM J GASTROENTEROL, V91, P1150
[4]   MOSAICISM IN VACUOLATING CYTOTOXIN ALLELES OF HELICOBACTER-PYLORI - ASSOCIATION OF SPECIFIC VACA TYPES WITH CYTOTOXIN PRODUCTION AND PEPTIC-ULCERATION [J].
ATHERTON, JC ;
CAO, P ;
PEEK, RM ;
TUMMURU, MKR ;
BLASER, MJ ;
COVER, TL .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (30) :17771-17777
[5]  
Atherton JC, 1998, BRIT MED BULL, V54, P105
[6]   Local pH elevation mediated by the intrabacterial urease of Helicobacter pylori cocultured with gastric cells [J].
Athmann, C ;
Zeng, NX ;
Kang, T ;
Marcus, EA ;
Scott, DR ;
Rektorschek, M ;
Buhmann, A ;
Melchers, K ;
Sachs, G .
JOURNAL OF CLINICAL INVESTIGATION, 2000, 106 (03) :339-347
[7]   Science, medicine, and the future -: Helicobacter pylori and gastric diseases [J].
Blaser, MJ .
BRITISH MEDICAL JOURNAL, 1998, 316 (7143) :1507-1510
[8]   Identification of an ovarian voltage-activated Na+-channel type:: Hints to involvement in luteolysis [J].
Bulling, A ;
Berg, FD ;
Berg, U ;
Duffy, DM ;
Stouffer, RL ;
Ojeda, SR ;
Gratzl, M ;
Mayerhofer, A .
MOLECULAR ENDOCRINOLOGY, 2000, 14 (07) :1064-1074
[9]   cag, a pathogenicity island of Helicobacter pylori, encodes type I-specific and disease-associated virulence factors [J].
Censini, S ;
Lange, C ;
Xiang, ZY ;
Crabtree, JE ;
Ghiara, P ;
Borodovsky, M ;
Rappuoli, R ;
Covacci, A .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (25) :14648-14653
[10]   Molecular mechanisms of TNF alpha cytotoxicity: Activation of NF-kappa B and nuclear translocation [J].
Claudio, E ;
Segade, F ;
Wrobel, K ;
Ramos, S ;
Bravo, R ;
Lazo, PS .
EXPERIMENTAL CELL RESEARCH, 1996, 224 (01) :63-71