The role of CB1 in intestinal permeability and inflammation
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 in...
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| Format: | Article |
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Federation of American Society of Experimental Biology
2017
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| Online Access: | https://eprints.nottingham.ac.uk/45054/ |
| _version_ | 1848797058484404224 |
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| author | Karwad, Mustafa A. Couch, Daniel G. Theophilidou, Elena Sarmad, Sarir Barrett, David A. Larvin, Michael Wright, Karen L. Lund, Jonathan N. O'Sullivan, Saoirse |
| author_facet | Karwad, Mustafa A. Couch, Daniel G. Theophilidou, Elena Sarmad, Sarir Barrett, David A. Larvin, Michael Wright, Karen L. Lund, Jonathan N. O'Sullivan, Saoirse |
| author_sort | Karwad, Mustafa A. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | 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-a and IFN-g and oxygen deprivation. Permeability changes were measured by transepithelial electrical resistance. The role of the CB1 receptor 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 permeability via CB1 (P < 0.0001) and decreased cytokine production. Basolateral application of JZL184 decreased permeability via CB1 (P < 0.0001). URB597 and JZL184 increased the enhanced (worsened) permeability caused by inflammation and hypoxia (P <0.0001 and P< 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 with ex vivo treatment with URB597 and JZL184, and was inhibited by a CB1 antagonist. The results of this study show that endogenous AEA and 2-AG production and CB1 activation play a key modulatory roles in normal intestinal mucosa permeability and in inflammatory and hypoxic conditions. |
| first_indexed | 2025-11-14T19:57:50Z |
| format | Article |
| id | nottingham-45054 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:57:50Z |
| publishDate | 2017 |
| publisher | Federation of American Society of Experimental Biology |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-450542024-08-15T15:22:21Z https://eprints.nottingham.ac.uk/45054/ The role of CB1 in intestinal permeability and inflammation Karwad, Mustafa A. Couch, Daniel G. Theophilidou, Elena Sarmad, Sarir Barrett, David A. Larvin, Michael Wright, Karen L. Lund, Jonathan N. O'Sullivan, Saoirse 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-a and IFN-g and oxygen deprivation. Permeability changes were measured by transepithelial electrical resistance. The role of the CB1 receptor 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 permeability via CB1 (P < 0.0001) and decreased cytokine production. Basolateral application of JZL184 decreased permeability via CB1 (P < 0.0001). URB597 and JZL184 increased the enhanced (worsened) permeability caused by inflammation and hypoxia (P <0.0001 and P< 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 with ex vivo treatment with URB597 and JZL184, and was inhibited by a CB1 antagonist. The results of this study show that endogenous AEA and 2-AG production and CB1 activation play a key modulatory roles in normal intestinal mucosa permeability and in inflammatory and hypoxic conditions. Federation of American Society of Experimental Biology 2017-04-12 Article PeerReviewed Karwad, Mustafa A., Couch, Daniel G., Theophilidou, Elena, Sarmad, Sarir, Barrett, David A., Larvin, Michael, Wright, Karen L., Lund, Jonathan N. and O'Sullivan, Saoirse (2017) The role of CB1 in intestinal permeability and inflammation. FASEB Journal, 31 (8). pp. 3267-3277. ISSN 1530-6860 gut endocannibinoids anandamide 2-ag http://www.fasebj.org/content/31/8/3267 doi:10.1096/fj.201601346R doi:10.1096/fj.201601346R |
| spellingShingle | gut endocannibinoids anandamide 2-ag Karwad, Mustafa A. Couch, Daniel G. Theophilidou, Elena Sarmad, Sarir Barrett, David A. Larvin, Michael Wright, Karen L. Lund, Jonathan N. O'Sullivan, Saoirse The role of CB1 in intestinal permeability and inflammation |
| title | The role of CB1 in intestinal permeability and inflammation |
| title_full | The role of CB1 in intestinal permeability and inflammation |
| title_fullStr | The role of CB1 in intestinal permeability and inflammation |
| title_full_unstemmed | The role of CB1 in intestinal permeability and inflammation |
| title_short | The role of CB1 in intestinal permeability and inflammation |
| title_sort | role of cb1 in intestinal permeability and inflammation |
| topic | gut endocannibinoids anandamide 2-ag |
| url | https://eprints.nottingham.ac.uk/45054/ https://eprints.nottingham.ac.uk/45054/ https://eprints.nottingham.ac.uk/45054/ |