Journal: Frontiers in Pharmacology

Article Title: Intestinal Epithelial Cell-Derived Extracellular Vesicles Modulate Hepatic Injury via the Gut-Liver Axis During Acute Alcohol Injury

doi: 10.3389/fphar.2020.603771

Figure Lengend Snippet: Acute alcohol injury disrupts the gut intestinal barrier in female and male mice. (A) Representative H&E staining was performed in paraffin colon sections of female and male mice treated with EtOH [6 g/kg] or PBS ( n = 5–7/group). (B) Quantification of the ZO-1 positive area was performed and graphed ( n = 4/group). (C) ZO-1 immunofluorescence staining was performed in colons of male and female mice. (D) FITC-dextran levels in serum from female and male mice was quantified as a measure of intestinal permeability ( n = 3–4/group). (E) MUCIN-2 immunofluorescence staining was performed in colons of female and male mice. (F) Quantification of the MUCIN-2 positive area was done and graphed ( n = 4–5/group; ** p < 0.01, **** p < 0.0001). In female mice, arrows mark mixed inflammation; double arrow, oedema in submucosa and a star, degeneration of crypt basal cells. In male mice, arrows mark mild hypertrophy in Goblet cells.

Article Snippet: Liver and colon from each mouse were preserved in cassettes in Tissue-Tek (Sakura Finetek U.S.A, Torrance, CA) at −80 C. Immunostainings for ZO-1 (Invitrogen, Paisley, FL), MUCIN-2 (Santa Cruz, Dallas, TX), TUNEL (Roche), and CD11b (BD, Madrid, Spain) were performed as previously described ( ).

Techniques: Staining, Immunofluorescence, Permeability

Journal: Frontiers in Pharmacology

Article Title: Intestinal Epithelial Cell-Derived Extracellular Vesicles Modulate Hepatic Injury via the Gut-Liver Axis During Acute Alcohol Injury

doi: 10.3389/fphar.2020.603771

Figure Lengend Snippet: Characterization of extracellular vesicles in vitro and in vivo . Isolated EVs from the supernatant of Caco-2 cells challenged with different concentrations of EtOH [0–100 mM] were measured by NTA, and (A) particles/ml and (B) size of EVs were represented. Isolated EVs were extracted from the serum of female and male mice treated with EtOH or PBS, and they were analyzed using NTA. (C) Particles/ml and (D) size of EVs were graphed ( n = 3/group; * p < 0.05, ** p < 0.01). (E) Schematic representation of the pathophysiological events that occur during acute alcohol exposure. Acute alcohol ingestion triggers damage to the intestinal epithelial barrier. Increased FITC-Dextran, disruption in tight junctions (ZO-1), and loss of mucosa (MUCIN-2) and alteration of the gut microbiota (increased Lactobacillus and decreased Lachnospiraceae) indicate damage to the intestinal epithelial barrier and gut dysbiosis. Translocation of microbial bacterial products through the leaky gut causes overexpression of tlr4 and intestinal-derived inflammation in the liver–presence of CD11b-positive cells, tnf-α , and il1β , and mild steatosis and lipid accumulation. Our in vitro experiments suggest that ethanol triggers the release of EVs by intestinal epithelial cells (IECs), which exert a deleterious effect on hepatocyte viability and lipid accumulation.

Article Snippet: Liver and colon from each mouse were preserved in cassettes in Tissue-Tek (Sakura Finetek U.S.A, Torrance, CA) at −80 C. Immunostainings for ZO-1 (Invitrogen, Paisley, FL), MUCIN-2 (Santa Cruz, Dallas, TX), TUNEL (Roche), and CD11b (BD, Madrid, Spain) were performed as previously described ( ).

Techniques: In Vitro, In Vivo, Isolation, Translocation Assay, Over Expression, Derivative Assay