RRC ID 65440
Author Yamada H, Aihara T, Okabe S.
Title Mechanism for Helicobacter pylori stimulation of interleukin-8 production in a gastric epithelial cell line (MKN 28): roles of mitogen-activated protein kinase and interleukin-1beta.
Journal Biochem Pharmacol
Abstract Although it is known that the pathogenic mechanism of Helicobacter pylori involves the stimulated production of interleukin-8 (IL-8) as an inflammatory mediator, the details of the pathway remain unclear. The role of mitogen-activated protein kinase (MAPK) in IL-8 production by H. pylori has been examined in an in vitro study. IL-8 mRNA expression in gastric epithelial cells (MKN 28) was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). IL-8 production was examined by ELISA. The activation of p38 MAPK was assessed by western blotting. Neither IL-8 mRNA nor activated p38 MAPK or p44/42 MAPK was detected in cells not treated with H. pylori. In contrast, incubation of cells with H. pylori, or IL-1beta, or both, clearly stimulated the expression of IL-8 mRNA within 60 min in a concentration-dependent manner. Phosphorylation of p38 MAPK and p44/p42 MAPK, as well as IL-8 production, occurred within 30 min and 24 hr after co-culturing MKN 28 cells with H. pylori and IL-1beta, respectively. Pretreatment of cells with MAPK inhibitors [1-[7-(4-fluorophenyl)-1,2,3,4-tetra-hydro-8-pyridylpyrazolo[5,1-c][1,2,4]triazin-2-yl]-2-phenylethanedione sulfate monohydrate (FR167653), 4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)imidazole (SB203580), or 2-(2'-amino-3'-methoxyphenyl)-oxanaphthalen-4-one (PD98059)] significantly inhibited IL-8 production stimulated by H. pylori or IL-1beta or both. The combination of H. pylori and IL-1beta additively stimulated IL-8 production. The additive effect of H. pylori and IL-1beta on IL-8 production was inhibited by treatment with a p38 MAPK inhibitor. It was revealed that the culturing of MKN 28 cells with H. pylori significantly stimulates IL-8 production to a degree sufficient for induction of neutrophil chemotaxis via activation of p38 and p44/42 MAPK.
Volume 61(12)
Pages 1595-604
Published 2001-6-15
DOI 10.1016/s0006-2952(01)00628-1
PII S0006295201006281
PMID 11377390
MeSH Activating Transcription Factor 2 Anti-Inflammatory Agents, Non-Steroidal / pharmacology Cyclic AMP Response Element-Binding Protein / metabolism Enzyme Activation Enzyme Inhibitors Helicobacter pylori / physiology* Humans Interleukin-1 / metabolism* Interleukin-8 / genetics Interleukin-8 / metabolism* Mitogen-Activated Protein Kinase 1 / metabolism Mitogen-Activated Protein Kinase 3 Mitogen-Activated Protein Kinases / antagonists & inhibitors Mitogen-Activated Protein Kinases / metabolism* Pyrazoles / pharmacology Pyridines / pharmacology RNA, Messenger / metabolism Time Factors Transcription Factors / metabolism Tumor Cells, Cultured p38 Mitogen-Activated Protein Kinases
IF 4.96
Human and Animal Cells MKN28(RCB1000)