title: Deletion of neutral endopeptidase exacerbates intestinal inflammation induced by Clostridium difficile toxin A.
abstract: Toxin A (TxA) of Clostridium difficile induces acute inflammation of the intestine initiated by release of substance P (SP) and activation of the neurokinin-1 receptor. However, the mechanisms that terminate this response are unknown. We determined whether the SP-degrading enzyme neutral endopeptidase (NEP, EC 3.4.24.11) terminates TxA-induced enteritis. We used both genetic deletion and pharmacological inhibition of NEP to test this hypothesis. In wild-type mice, instillation of TxA (0.5-5 microg) into ileal loops for 3 h dose dependently increased ileal fluid secretion, stimulated granulocyte transmigration determined by myeloperoxidase activity, and caused histological damage characterized by depletion of enterocytes, edema, and neutrophil accumulation. Deletion of NEP reduced the threshold secretory and inflammatory dose of TxA and exacerbated the inflammatory responses by more than twofold. This exacerbated inflammation was prevented by pretreatment with recombinant NEP. Conversely, pretreatment of wild-type mice with the NEP inhibitor phosphoramidon exacerbated enteritis. Thus NEP terminates enteritis induced by C. difficile TxA, underlying the importance of SP degradation in limiting neurogenic inflammation.
authors.affiliation: Department of Surgery, University of California, San Francisco, California 94143, USA.
authors.forename: K S
authors.initials: KS
authors.lastname: Kirkwood
authors.forename: N W
authors.initials: NW
authors.lastname: Bunnett
authors.forename: J
authors.initials: J
authors.lastname: Maa
authors.forename: I
authors.initials: I
authors.lastname: Castagliolo
authors.forename: B
authors.initials: B
authors.lastname: Liu
authors.forename: N
authors.initials: N
authors.lastname: Gerard
authors.forename: J
authors.initials: J
authors.lastname: Zacks
authors.forename: C
authors.initials: C
authors.lastname: Pothoulakis
authors.forename: E F
authors.initials: EF
authors.lastname: Grady
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D001427:Bacterial Toxins / Q000494:pharmacology*; D004761:Enterocolitis, Pseudomembranous / Q000276:immunology / Q000378:metabolism / Q000473:pathology*; D004768:Enterotoxins / Q000494:pharmacology*; D006020:Glycopeptides / Q000494:pharmacology; D006098:Granulocytes / Q000276:immunology; D007413:Intestinal Mucosa / Q000473:pathology; D007419:Intestinal Secretions / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D015260:Neprilysin / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology
chemical_list: D000893:Anti-Inflammatory Agents; D001427:Bacterial Toxins; D004768:Enterotoxins; D006020:Glycopeptides; D011994:Recombinant Proteins; C041348:tcdA protein, Clostridium difficile; D015260:Neprilysin; C008890:phosphoramidon