title: Dysregulation of microRNA biogenesis in the small intestine after ethanol and burn injury.
abstract: Ethanol exposure at the time of burn injury is a major contributor to post-burn pathogenesis. Many of the adverse effects associated with ethanol and burn injury are linked to an impaired intestinal barrier. The combined insult causes intestinal inflammation, resulting in tissue damage, altered tight junction expression, and increased intestinal permeability. MicroRNAs play a critical role in maintaining intestinal homeostasis including intestinal inflammation and barrier function. Specifically, miR-150 regulates inflammatory mediators which can contribute to gut barrier disruption. The present study examined whether ethanol and burn injury alter expression of microRNA processing enzymes (Drosha, Dicer, and Argonaute-2) and miR-150 in the small intestine. Male mice were gavaged with ethanol (~2.9g/kg) 4h prior to receiving a ~12.5% total body surface area full thickness burn. One or three days after injury, mice were euthanized and small intestinal epithelial cells (IECs) were isolated and analyzed for expression of microRNA biogenesis components and miR-150. Dicer mRNA and protein levels were not changed following the combined insult. Drosha and Argonaute-2 mRNA and protein levels were significantly reduced in IECs one day after injury; which accompanied reduced miR-150 expression. To further determine the role of miR-150 in intestinal inflammation, young adult mouse colonocytes were transfected with a miR-150 plasmid and stimulated with LPS (100ng/ml). miR-150 overexpression significantly reduced IL-6 and KC protein levels compared to vector control cells challenged with LPS. These results suggest that altered microRNA biogenesis and associated decrease in miR-150 likely contribute to increased intestinal inflammation following ethanol and burn injury.
authors.affiliation: Alcohol Research Program, Burn and Shock Trauma Research Institute, Department of Surgery, Loyola University Chicago Health Sciences Campus, Maywood, IL 60153, USA.; Integrative Cell Biology Program, Loyola University Chicago Health Sciences Campus, Maywood, IL 60153, USA. Electronic address: nmorris3@luc.edu.
authors.forename: Niya L
authors.initials: NL
authors.lastname: Morris
authors.affiliation: Alcohol Research Program, Burn and Shock Trauma Research Institute, Department of Surgery, Loyola University Chicago Health Sciences Campus, Maywood, IL 60153, USA.; Integrative Cell Biology Program, Loyola University Chicago Health Sciences Campus, Maywood, IL 60153, USA. Electronic address: adhammer@luc.edu.
authors.forename: Adam M
authors.initials: AM
authors.lastname: Hammer
authors.affiliation: Alcohol Research Program, Burn and Shock Trauma Research Institute, Department of Surgery, Loyola University Chicago Health Sciences Campus, Maywood, IL 60153, USA.; Integrative Cell Biology Program, Loyola University Chicago Health Sciences Campus, Maywood, IL 60153, USA. Electronic address: acannon@luc.edu.
authors.forename: Abigail R
authors.initials: AR
authors.lastname: Cannon
authors.affiliation: Alcohol Research Program, Burn and Shock Trauma Research Institute, Department of Surgery, Loyola University Chicago Health Sciences Campus, Maywood, IL 60153, USA.. Electronic address: rgagnon1@luc.edu.
authors.forename: Robin C
authors.initials: RC
authors.lastname: Gagnon
authors.affiliation: Alcohol Research Program, Burn and Shock Trauma Research Institute, Department of Surgery, Loyola University Chicago Health Sciences Campus, Maywood, IL 60153, USA.. Electronic address: xiali@luc.edu.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Li
authors.affiliation: Alcohol Research Program, Burn and Shock Trauma Research Institute, Department of Surgery, Loyola University Chicago Health Sciences Campus, Maywood, IL 60153, USA.; Integrative Cell Biology Program, Loyola University Chicago Health Sciences Campus, Maywood, IL 60153, USA; Department of Microbiology and Immunology, Loyola University Chicago Health Sciences Campus, Maywood, IL 60153, USA. Electronic address: mchoudhry@luc.edu.
authors.forename: Mashkoor A
authors.initials: MA
authors.lastname: Choudhry
mesh_terms: D000818:Animals; D060565:Argonaute Proteins / Q000276:immunology / Q000378:metabolism; D002056:Burns / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D054360:Chemokine CXCL1 / Q000276:immunology / Q000378:metabolism; D053487:DEAD-box RNA Helicases / Q000276:immunology / Q000378:metabolism; D000431:Ethanol / Q000009:adverse effects* / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000276:immunology*; D018836:Inflammation Mediators / Q000276:immunology / Q000378:metabolism; D015850:Interleukin-6 / Q000276:immunology / Q000378:metabolism; D007413:Intestinal Mucosa / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D007421:Intestine, Small / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000276:immunology* / Q000378:metabolism; D012333:RNA, Messenger / Q000276:immunology / Q000378:metabolism; D043244:Ribonuclease III / Q000276:immunology / Q000378:metabolism
chemical_list: C526735:Ago2 protein, mouse; D060565:Argonaute Proteins; D054360:Chemokine CXCL1; C475895:Cxcl1 protein, mouse; D018836:Inflammation Mediators; D015850:Interleukin-6; D035683:MicroRNAs; C529101:Mirn150 microRNA, mouse; D012333:RNA, Messenger; C577686:interleukin-6, mouse; D000431:Ethanol; C509781:Dicer1 protein, mouse; D043244:Ribonuclease III; D053487:DEAD-box RNA Helicases
keywords: Alcohol; Gut; Inflammation; MicroRNA processing; Trauma; miRs