Role of defective methylation reactions in ethanol-induced dysregulation of intestinal barrier integrity.
Alcoholic liver disease (ALD) is a major healthcare challenge worldwide. Emerging evidence reveals that ethanol administration disrupts the intestinal epithelial tight junction (TJ) complex; this defect allows for the paracellular translocation of gut-derived pathogenic molecules to reach the liver to cause inflammation and progressive liver injury. We have previously demonstrated a causative role of impairments in liver transmethylation reactions in the pathogenesis of ALD. We have further shown that treatment with betaine, a methylation agent that normalizes liver methylation potential, can attenuate ethanol-induced liver injury. Herein, we explored whether alterations in methylation reactions play a causative role in disrupting intestinal mucosal barrier function by employing an intestinal epithelial cell line. Monolayers of Caco-2 cells were exposed to ethanol or a-pan methylation reaction inhibitor, tubercidin, in the presence and absence of betaine. The structural and functional integrity of intestinal epithelial barrier was then examined. We observed that exposure to either ethanol or tubercidin disrupted TJ integrity and function by decreasing the localization of TJ protein occludin-1 to the intracellular junctions, reducing transepithelial electrical resistance and increasing dextran influx. All these detrimental effects of ethanol and tubercidin were attenuated by co-treatment with betaine. We further show that the mechanism of betaine protection was through BHMT-mediated catalysis. Collectively, our data suggest a novel mechanism for alcohol-induced gut leakiness and identifies the importance of normal methylation reactions in maintaining TJ integrity. We also propose betaine as a potential therapeutic option for leaky gut in alcohol-consuming patients who are at the risk of developing ALD.
Research Service, Veterans Affairs Nebraska-Western Iowa Health Care System, Omaha, NE 68105, USA; Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE 68198, USA.
Paul G
PG
Thomes
Research Service, Veterans Affairs Nebraska-Western Iowa Health Care System, Omaha, NE 68105, USA; Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE 68198, USA.
Natalia A
NA
Osna
Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE 68198, USA.
Sarah M
SM
Bligh
Research Service, Veterans Affairs Nebraska-Western Iowa Health Care System, Omaha, NE 68105, USA; Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE 68198, USA.
Dean J
DJ
Tuma
Research Service, Veterans Affairs Nebraska-Western Iowa Health Care System, Omaha, NE 68105, USA; Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE 68198, USA; Department of Biochemistry & Molecular Biology, University of Nebraska Medical Center, Omaha, NE 68198, USA. Electronic address: kkharbanda@unmc.edu.
Kusum K
KK
Kharbanda
D015153:Blotting, Western; D000431:Ethanol / Q000494:pharmacology*; D006801:Humans; D007413:Intestinal Mucosa / Q000187:drug effects* / Q000503:physiopathology; D008745:Methylation
D000431:Ethanol
Betaine; Caco-2; Ethanol; Methylation; Occludin-1; Tubercidin