title: Phorbol myristate acetate ex vivo model of enhanced colonic epithelial permeability. Reactive oxygen metabolite and protease independence.
abstract: The initiating mechanisms involved in colonic injury are currently unknown. The goal of the current study was to examine the role of the inflammatory mediators reactive oxygen metabolites and proteases in an ex vivo model of selective epithelial permeability. Rats were prepared with exteriorized colonic chambers to which the protein kinase C (PKC) activator phorbol myristate acetate (PMA) was added in doses ranging from 5 to 800 micrograms. PMA caused a dose-dependent transient increase in epithelial permeability, but had no significant effect on microvascular permeability. There was no accumulation of neutrophils and no apparent histological changes. PMA acts via a PKC-dependent mechanism, as assessed using the PKC-inactive phorbol analog 4 alpha-phorbol didecanoate, and the response is tachyphylactic. The mechanism is independent of reactive oxygen metabolites and proteases, as shown by the lack of effect of the free radical scavengers superoxide dismutase and catalase and the general serine protease inhibitor soybean trypsin inhibitor. The classic inflammatory process does not appear to be involved in the PMA-induced epithelial permeability changes. This finding suggests that noninflammatory mechanisms may regulate the increased epithelial permeability induced by PMA. Further study to elucidate these mechanisms is of importance for understanding both normal gastrointestinal physiology and initiation of pathology.
authors.affiliation: Department of Physiology, Queen's University, Hotel Dieu Hospital, Kingston, Ontario, Canada.
authors.forename: M C
authors.initials: MC
authors.lastname: Berin
authors.forename: M G
authors.initials: MG
authors.lastname: Buell
mesh_terms: D000208:Acute Disease; D000818:Animals; D002463:Cell Membrane Permeability / Q000187:drug effects*; D003092:Colitis / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology; D003106:Colon / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D018836:Inflammation Mediators / Q000378:metabolism; D008297:Male; D011493:Protein Kinase C / Q000819:agonists*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000187:drug effects* / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000493:pharmacokinetics / Q000494:pharmacology*
chemical_list: D018836:Inflammation Mediators; D017382:Reactive Oxygen Species; D011493:Protein Kinase C; D012697:Serine Endopeptidases; D013755:Tetradecanoylphorbol Acetate