title: TNFalpha processing enzyme inhibitors prevent aspirin-induced TNFalpha release and protect against gastric mucosal injury in rats.
abstract: BACKGROUND
  Although previous studies indicate that prevention of tumour necrosis factor alpha (TNFalpha) release protects against NSAID-induced gastric mucosal injury, intracellular pathways by which aspirin causes TNFalpha release are unknown. TNFalpha is synthesized as a precursor which is proteolytically cleaved by a specific converting enzyme, TACE, to release the mature cytokine. TACE inhibitors prevent TNFalpha release and protect against TNFalpha-mediated disease.
  
  
  OBJECTIVE
  To investigate: (i) molecular events that regulate TNFalpha secretion in response to aspirin in vivo and in vitro; (ii) whether TNFalpha secretion inhibitors prevent aspirin-induced TNFalpha release and protect against gastric mucosal damage; and (iii) whether TNFalpha exerts a direct cytotoxic effect on gastric epithelial cells.
  
  
  METHODS
  In vitro studies were carried out on mouse macrophages and rat gastric mucosal cells. Gastric mucosal damage was induced in rats by oral administration of 300 mg/kg aspirin. TNFalpha cytotoxicity on gastric mucosal cells was examined by treating rats with lipopolysaccharide to release TNFalpha or by incubating dispersed gastric mucosal cells with increasing concentrations of TNFalpha.
  
  
  RESULTS
  Aspirin increases intracellular calcium (Ca2+) levels and causes a time and concentration dependent increase in macrophage TNFalpha mRNA accumulation and cytokine release. Agents that cause Ca2+ mobilization with a receptor-independent mechanism, such as ionomycin and thapsigargin, stimulate TNFalpha release. Incubating the macrophages in a Ca2+ free medium inhibited TNFalpha secretion. Agents that prevent TNFalpha mRNA transcription, e.g. lisophylline, PGE2, interleukin-10 and 8-BrcAMP, or TACE inhibitors, e.g. EDTA, TAPI-2 and BB-3103, inhibit TNFalpha release and protect rats against gastric mucosal injury induced by oral administration of aspirin. TNFalpha exerts a direct cytotoxic effect on gastric epithelial cells as demonstrated by the reduced viability observed in gastric mucosal cells prepared from rats treated with lipopolysaccharide, or directly incubated with increasing concentrations of TNFalpha.
  
  
  CONCLUSIONS
  (i) Aspirin directly stimulates TNFalpha gene transcription; (ii) TACE inhibitors protect against aspirin-induced gastric mucosal injury; and (iii) TNFalpha exerts a direct cytotoxic effect on gastric epithelial cells.
authors.affiliation: Clinica di Gastroenterologia ed Endoscopia Digestiva, Dipartimento di Medicina Clinica, Farmacologia e Patologia, Università degli Studi di Perugia, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Fiorucci
authors.forename: E
authors.initials: E
authors.lastname: Antonelli
authors.forename: G
authors.initials: G
authors.lastname: Migliorati
authors.forename: L
authors.initials: L
authors.lastname: Santucci
authors.forename: O
authors.initials: O
authors.lastname: Morelli
authors.forename: B
authors.initials: B
authors.lastname: Federici
authors.forename: A
authors.initials: A
authors.lastname: Morelli
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology; D001241:Aspirin / Q000494:pharmacology*; D015152:Blotting, Northern; D002478:Cells, Cultured; D004492:Edetic Acid / Q000494:pharmacology; D005753:Gastric Mucosa / Q000187:drug effects*; D006877:Hydroxamic Acids / Q000494:pharmacology; D008264:Macrophages / Q000378:metabolism; D008297:Male; D051379:Mice; D010431:Pentoxifylline / Q000031:analogs & derivatives / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; C108282:BB 3103; D006877:Hydroxamic Acids; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; C111875:lisofylline 4,5-diol; D004492:Edetic Acid; D001241:Aspirin; D010431:Pentoxifylline