Inhibition of proinflammatory cytokine expression by NF-kappaB (p65) antisense oligonucleotide in Helicobacter pylori-infected mice.
BACKGROUND
Helicobacter pylori induces the expression of proinflammatory cytokines in vitro by activating nuclear factor-kappaB, a transcriptional regulator. However, it has not been clarified whether H. pylori-induced proinflammatory cytokines are also mediated through nuclear factor-kappaB in vivo. The aim of this study was to evaluate the role of nuclear factor-kappaB on the expressions of proinflammatory cytokines in H. pylori-infected mice.


METHODS
We evaluated nuclear factor-kappaB (p65) activation in the H. pylori-infected gastric mucosa of mice by immunofluorescent staining using antip65 polyclonal antibody, and the expressions of proinflammatory cytokines with inhibition of nuclear factor-kappaB pathway by using phosphorothioate antisense and sense oligonucleotide against the nuclear factor-kappaB (p65).


RESULTS
In the H. pylori-infected gastric mucosa of mice, immunofluorescent staining using antip65 polyclonal antibody showed nuclear factor-kappaB (p65) activation, which was particularly localized to epithelial cells. Tumor necrosis factor-alpha and interleukin-1beta concentrations in gastric mucosa by enzyme-linked immunosorbent assay (ELISA) were elevated in the infected group versus the uninfected group. Pretreatment with nuclear factor-kappaB (p65) antisense oligonucleotide inhibited the activation of nuclear factor-kappaB and the expressions of tumor necrosis factor-alpha and interleukin-1beta in H. pylori-infected gastric mucosa. Sense oligonucleotide did not influence on the expression of proinflammatory cytokines.


CONCLUSIONS
H. pylori infection was found to activate the expressions of proinflammatory cytokines via nuclear factor-kappaB in vivo, and this may play an important role in the initiation of H. pylori-induced gastric inflammation.
Kim
Sang Gyun
SG
Department of Internal Medicine and Liver Research Institute, Seoul National University College of Medicine, Chongno-Gu, Seoul, Korea.
Kim
Joo Sung
JS
Kim
Jung Mogg
JM
Chae Jung
Hyun
H
Sung Song
In
I
D000818:Animals; D016207:Cytokines / Q000037:antagonists & inhibitors* / Q000378:metabolism; D005260:Female; D005753:Gastric Mucosa / Q000187:drug effects / Q000276:immunology* / Q000382:microbiology / Q000503:physiopathology; D016481:Helicobacter Infections / Q000188:drug therapy* / Q000276:immunology / Q000503:physiopathology; D016480:Helicobacter pylori / Q000276:immunology* / Q000472:pathogenicity; D007375:Interleukin-1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000494:pharmacology* / Q000627:therapeutic use; D051996:Transcription Factor RelA / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000378:metabolism
D016207:Cytokines; D007375:Interleukin-1; D016376:Oligonucleotides, Antisense; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha