IL-1beta-induced apoptosis in rat gastric enterochromaffin-like cells is mediated by iNOS, NF-kappaB, and Bax protein

Gastroenterology. 2000 Mar;118(3):515-24. doi: 10.1016/s0016-5085(00)70257-5.


Background & aims: Enterochromaffin-like (ECL) cells are histamine-containing endocrine cells in the gastric mucosa. Previous studies have shown that the proinflammatory cytokine interleukin (IL)-1beta present during chronic gastritis inhibits histamine synthesis in ECL cells and leads to sustained functional impairment. This study investigated the effects of IL-1beta on ECL cell apoptosis and the related signal-transduction mechanisms.

Methods: ECL cells were isolated by pronase digestion and a combination of elutriation, gradient centrifugation, and 48-hour culture (purity >/=90%). Apoptosis was measured by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling reaction and cell death detection enzyme-linked immunosorbent assay.

Results: IL-1beta (100 pg/mL) increased the rate of programmed cell death 2-3 fold in ECL cells after 24 hours of incubation (total of 12%-14%). This effect was completely inhibited by the NF-kappaB inhibitor, proteasome inhibitor I, and the nitric oxide synthase inhibitor (iNOS) N(G)-monomethyl-L-arginine (10(-4) mol/L), but not by the caspase 3 inhibitor, Asp-Glu-Val-Asp-CHO. Western blot analysis, reverse-transcription polymerase chain reaction (PCR), and in situ PCR showed that IL-1beta induced gene expression (after 2-4 hours) and protein synthesis (6-18 hours) of the iNOS isoform in ECL cells. Bax protein expression was increased in response to IL-1beta. In contrast, bcl-2 gene expression was increased in response to basic fibroblast growth factor, which has been shown to counteract IL-1beta- induced apoptosis.

Conclusions: These data suggest that IL-1beta induces programmed cell death in isolated rat ECL cells via activation of NF-kappaB, iNOS, and the Bax protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Apoptosis / physiology*
  • Caspase 3
  • Caspase Inhibitors
  • Cysteine Endopeptidases / drug effects
  • Enterochromaffin Cells / metabolism
  • Enterochromaffin Cells / physiology*
  • Enzyme Inhibitors / pharmacology
  • Female
  • Fibroblast Growth Factor 2 / pharmacology
  • Gastric Mucosa / cytology
  • Gastric Mucosa / metabolism
  • Gastric Mucosa / physiology*
  • Histidine Decarboxylase / genetics
  • Interleukin-1 / pharmacology*
  • Multienzyme Complexes / drug effects
  • NF-kappa B / antagonists & inhibitors
  • NF-kappa B / physiology*
  • Nitric Oxide Synthase / antagonists & inhibitors
  • Nitric Oxide Synthase / genetics
  • Nitric Oxide Synthase / metabolism
  • Nitric Oxide Synthase / physiology*
  • Nitric Oxide Synthase Type II
  • Oligopeptides / pharmacology
  • Proteasome Endopeptidase Complex
  • Proto-Oncogene Proteins / physiology*
  • Proto-Oncogene Proteins c-bcl-2*
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • bcl-2-Associated X Protein
  • omega-N-Methylarginine / pharmacology


  • Bax protein, rat
  • Caspase Inhibitors
  • Enzyme Inhibitors
  • Interleukin-1
  • Multienzyme Complexes
  • NF-kappa B
  • Oligopeptides
  • Proto-Oncogene Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Messenger
  • aspartyl-glutamyl-valyl-aspartal
  • bcl-2-Associated X Protein
  • Fibroblast Growth Factor 2
  • omega-N-Methylarginine
  • Nitric Oxide Synthase
  • Nitric Oxide Synthase Type II
  • Nos2 protein, rat
  • Casp3 protein, rat
  • Caspase 3
  • Cysteine Endopeptidases
  • Proteasome Endopeptidase Complex
  • Histidine Decarboxylase