title: Defects in 15-HETE Production and Control of Epithelial Permeability by Human Enteric Glial Cells From Patients With Crohn's Disease.
abstract: OBJECTIVE
  Enteric glial cells (EGCs) produce soluble mediators that regulate homeostasis and permeability of the intestinal epithelial barrier (IEB). We investigated the profile of polyunsaturated fatty acid (PUFA) metabolites produced by EGCs from rats and from patients with Crohn's disease (CD), compared with controls, along with the ability of one of these metabolites, 15-hydroxyeicosatetraenoic acid (15-HETE), to regulate the permeability of the IEB.
  
  
  METHODS
  We isolated EGCs from male Sprague-Dawley rats, intestinal resections of 6 patients with CD, and uninflamed healthy areas of intestinal tissue from 6 patients who underwent surgery for colorectal cancer (controls). EGC-conditioned media was analyzed by high-sensitivity liquid-chromatography tandem mass spectrometry to determine PUFA signatures. We used immunostaining to identify 15-HETE-producing enzymes in EGCs and tissues. The effects of human EGCs and 15-HETE on permeability and transepithelial electrical resistance of the IEB were measured using Caco-2 cells; effects on signal transduction proteins were measured with immunoblots. Levels of proteins were reduced in Caco-2 cells using short-hairpin RNAs or proteins were inhibited pharmacologically. Rats were given intraperitoneal injections of 15-HETE or an inhibitor of 15-lipoxygenase (the enzyme that produces 15-HETE); colons were collected and permeability was measured.
  
  
  RESULTS
  EGCs expressed 15-lipoxygenase-2 and produced high levels of 15-HETE, which increased IEB resistance and reduced IEB permeability. 15-HETE production was reduced in EGCs from patients with CD compared with controls. EGCs from patients with CD were unable to reduce the permeability of the IEB; the addition of 15-HETE restored permeability to levels of control tissues. Inhibiting 15-HETE production in rats increased the permeability of the IEB in colon tissues. We found that 15-HETE regulates IEB permeability by inhibiting an adenosine monophosphate-activated protein kinase and increasing expression of zonula occludens-1.
  
  
  CONCLUSIONS
  Enteric glial cells from patients with CD have reduced production of 15-HETE, which controls IEB permeability by inhibiting adenosine monophosphate-activated protein kinase and increasing expression of zonula occludens-1.
authors.affiliation: INSERM, UMR913, Nantes, France; Nantes University, Nantes, France; Institut des Maladies de l'Appareil Digestif, IMAD, CHU Nantes, Hopital Hôtel-Dieu, Nantes, France; Centre de Recherche en Nutrition Humaine, Nantes, France.
authors.forename: Camille
authors.initials: C
authors.lastname: Pochard
authors.affiliation: INSERM, UMR913, Nantes, France; Nantes University, Nantes, France; Institut des Maladies de l'Appareil Digestif, IMAD, CHU Nantes, Hopital Hôtel-Dieu, Nantes, France; Centre de Recherche en Nutrition Humaine, Nantes, France.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Coquenlorge
authors.affiliation: INSERM, UMR913, Nantes, France; Nantes University, Nantes, France; Institut des Maladies de l'Appareil Digestif, IMAD, CHU Nantes, Hopital Hôtel-Dieu, Nantes, France; Centre de Recherche en Nutrition Humaine, Nantes, France.
authors.forename: Julie
authors.initials: J
authors.lastname: Jaulin
authors.affiliation: INSERM UMR1043, Toulouse, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Cenac
authors.affiliation: INSERM UMR1043, Toulouse, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Vergnolle
authors.affiliation: INSERM, UMR913, Nantes, France; Nantes University, Nantes, France; Institut des Maladies de l'Appareil Digestif, IMAD, CHU Nantes, Hopital Hôtel-Dieu, Nantes, France; Centre de Recherche en Nutrition Humaine, Nantes, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Meurette
authors.affiliation: INSERM, UMR913, Nantes, France; Institut des Maladies de l'Appareil Digestif, IMAD, CHU Nantes, Hopital Hôtel-Dieu, Nantes, France; Centre de Recherche en Nutrition Humaine, Nantes, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Freyssinet
authors.affiliation: INSERM, UMR913, Nantes, France; Nantes University, Nantes, France; Institut des Maladies de l'Appareil Digestif, IMAD, CHU Nantes, Hopital Hôtel-Dieu, Nantes, France; Centre de Recherche en Nutrition Humaine, Nantes, France. Electronic address: Michel.Neunlist@univ-nantes.fr.
authors.forename: Michel
authors.initials: M
authors.lastname: Neunlist
authors.affiliation: INSERM, UMR913, Nantes, France; Nantes University, Nantes, France; Institut des Maladies de l'Appareil Digestif, IMAD, CHU Nantes, Hopital Hôtel-Dieu, Nantes, France; Centre de Recherche en Nutrition Humaine, Nantes, France. Electronic address: malvyne.derkinderen@univ-nantes.fr.
authors.forename: Malvyne
authors.initials: M
authors.lastname: Rolli-Derkinderen
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western; D018938:Caco-2 Cells / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000502:physiology*; D002478:Cells, Cultured; D003424:Crohn Disease / Q000378:metabolism*; D004195:Disease Models, Animal; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000378:metabolism*; D007150:Immunohistochemistry; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism; D008297:Male; D009457:Neuroglia / Q000378:metabolism*; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D012016:Reference Values
chemical_list: D006893:Hydroxyeicosatetraenoic Acids
keywords: Arachidonic Acid; Enteric Nervous System; IBD; Inflammatory Bowel Disease