title: The role of CB1in intestinal permeability and inflammation.
abstract: The endocannabinoid system has previously been shown to play a role in the permeability and inflammatory response of the human gut. The goal of our study was to determine the effects of endogenous anandamide (AEA) and 2-arachidonoyl glycerol (2-AG) on the permeability and inflammatory response of intestinal epithelium under normal, inflammatory, and hypoxic conditions. Human intestinal mucosa was modeled using Caco-2 cells. Human tissue was collected from planned colorectal resections. Accumulation of AEA and 2-AG was achieved by inhibiting their metabolizing enzymes URB597 (a fatty acid amide hydrolase inhibitor) and JZL184 (a monoacylglycerol lipase inhibitor). Inflammation and ischemia were simulated with TNF-α and IFN-γ and oxygen deprivation. Permeability changes were measured by transepithelial electrical resistance. The role of the CB1receptor was explored using CB1-knockdown (CB1Kd) intestinal epithelial cells. Endocannabinoid levels were measured using liquid chromatography-mass spectrometry. Cytokine secretion was measured using multiplex and ELISA. URB597 and JZL184 caused a concentration-dependent increase in permeabilityviaCB1(P< 0.0001) and decreased cytokine production. Basolateral application of JZL184 decreased permeabilityviaCB1(P< 0.0001). URB597 and JZL184 increased the enhanced (worsened) permeability caused by inflammation and hypoxia (P< 0.0001 andP< 0.05). CB1Kd cells showed reduced permeability response to inflammation (P< 0.01) but not hypoxia. 2-AG levels were increased in response to inflammation and hypoxia in Caco-2 cells. In human mucosal tissue, inflammation increased the secretion of granulocyte macrophage-colony stimulating factor, IL-12, -13, and -15, which was prevented withex vivotreatment with URB597 and JZL184, and was inhibited by a CB1antagonist. The results of this study show that endogenous AEA and 2-AG production and CB1activation play a key modulatory roles in normal intestinal mucosa permeability and in inflammatory and hypoxic conditions.-Karwad, M. A., Couch, D. G., Theophilidou, E., Sarmad, S., Barrett, D. A., Larvin, M., Wright, K. L., Lund, J. N., O'Sullivan, S. E. The role of CB1in intestinal permeability and inflammation.
authors.affiliation: School of Medicine, Royal Derby Hospital, University of Nottingham, Nottingham, United Kingdom.
authors.forename: Mustafa A
authors.initials: MA
authors.lastname: Karwad
authors.affiliation: School of Medicine, Royal Derby Hospital, University of Nottingham, Nottingham, United Kingdom.
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Couch
authors.affiliation: School of Medicine, Royal Derby Hospital, University of Nottingham, Nottingham, United Kingdom.
authors.forename: Elena
authors.initials: E
authors.lastname: Theophilidou
authors.affiliation: Centre for Analytical Bioscience, School of Pharmacy, University of Nottingham, Nottingham, United Kingdom.
authors.forename: Sarir
authors.initials: S
authors.lastname: Sarmad
authors.affiliation: Centre for Analytical Bioscience, School of Pharmacy, University of Nottingham, Nottingham, United Kingdom.
authors.forename: David A
authors.initials: DA
authors.lastname: Barrett
authors.affiliation: Graduate Entry Medical School, University of Limerick, Limerick, Ireland; University of Limerick, Limerick, Ireland.
authors.forename: Michael
authors.initials: M
authors.lastname: Larvin
authors.affiliation: Division of Biomedical and Life Sciences, Faculty of Health and Medicine, Lancaster University, Lancaster, United Kingdom.
authors.forename: Karen L
authors.initials: KL
authors.lastname: Wright
authors.affiliation: School of Medicine, Royal Derby Hospital, University of Nottingham, Nottingham, United Kingdom.
authors.forename: Jonathan N
authors.initials: JN
authors.lastname: Lund
authors.affiliation: School of Medicine, Royal Derby Hospital, University of Nottingham, Nottingham, United Kingdom; couch27@gmail.com.
authors.forename: Saoirse E
authors.initials: SE
authors.lastname: O'Sullivan
mesh_terms: D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism; D001095:Arachidonic Acids / Q000378:metabolism*; D001549:Benzamides / Q000494:pharmacology; D052117:Benzodioxoles / Q000494:pharmacology; D018938:Caco-2 Cells; D002219:Carbamates / Q000494:pharmacology; D015179:Colorectal Neoplasms / Q000378:metabolism; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D017097:Electric Impedance; D063388:Endocannabinoids / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology; D005989:Glycerides / Q000378:metabolism*; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D007413:Intestinal Mucosa / Q000378:metabolism / Q000473:pathology; D007422:Intestines / Q000473:pathology / Q000502:physiology*; D008994:Monoacylglycerol Lipases / Q000037:antagonists & inhibitors / Q000378:metabolism; D010101:Oxygen Consumption; D010539:Permeability; D010880:Piperidines / Q000494:pharmacology; D053284:Polyunsaturated Alkamides / Q000378:metabolism*; D043884:Receptor, Cannabinoid, CB1 / Q000235:genetics / Q000378:metabolism*; D046509:Tissue Culture Techniques; D000099252:Fatty Acid Amide Hydrolases
chemical_list: D000581:Amidohydrolases; D001095:Arachidonic Acids; D001549:Benzamides; D052117:Benzodioxoles; D002219:Carbamates; D016207:Cytokines; D063388:Endocannabinoids; D005989:Glycerides; D008994:Monoacylglycerol Lipases; D010880:Piperidines; D053284:Polyunsaturated Alkamides; D043884:Receptor, Cannabinoid, CB1; D000099252:Fatty Acid Amide Hydrolases; C534333:JZL 184; C500528:cyclohexyl carbamic acid 3'-carbamoylbiphenyl-3-yl ester; C094503:glyceryl 2-arachidonate; C078814:anandamide
keywords: 2-ag; anandamide; endocannabinoids; gut