IL-1beta-induced apoptosis in rat gastric enterochromaffin-like cells is mediated by iNOS, NF-kappaB, and Bax protein
| Autor: | Markus Gerhard, Meinhard Classen, Nina Neumayer, Sabine Mahr, Christian Prinz |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
endocrine system
Programmed cell death Proteasome Endopeptidase Complex medicine.medical_treatment Nitric Oxide Synthase Type II Caspase 3 Apoptosis Biology Histidine Decarboxylase Rats Sprague-Dawley Bcl-2-associated X protein Multienzyme Complexes Proto-Oncogene Proteins medicine Enterochromaffin Cells Animals RNA Messenger Enterochromaffin-like cell Enzyme Inhibitors bcl-2-Associated X Protein TUNEL assay omega-N-Methylarginine Hepatology Gastroenterology NF-kappa B Molecular biology Caspase Inhibitors Rats Cysteine Endopeptidases Cytokine Proto-Oncogene Proteins c-bcl-2 Gastric Mucosa biology.protein Proteasome inhibitor Female Fibroblast Growth Factor 2 Nitric Oxide Synthase Oligopeptides medicine.drug Interleukin-1 |
| Zdroj: | Europe PubMed Central |
| ISSN: | 0016-5085 |
| Popis: | 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)-1β present during chronic gastritis inhibits histamine synthesis in ECL cells and leads to sustained functional impairment. This study investigated the effects of IL-1β 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-1β (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-κB 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-1β 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-1β. In contrast, bcl-2 gene expression was increased in response to basic fibroblast growth factor, which has been shown to counteract IL-1β– induced apoptosis. Conclusions: These data suggest that IL-1β induces programmed cell death in isolated rat ECL cells via activation of NF-κB, iNOS, and the Bax protein. GASTROENTEROLOGY 2000;118:515-524 |
| Databáze: | OpenAIRE |
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