Dual therapy with zinc acetate and rifaximin prevents from ethanol-induced liver fibrosis by maintaining intestinal barrier integrity

Figure 5 Zinc acetate and rifaximin on intestinal barrier function in alcoholic liver disease mice. A: Representative microphotographs of ileum sections stained with hematoxylin-eosin (upper), zonula occludens-1 (ZO-1) (middle) and Occludin (lower) in the experimental groups. Boxes are selected regions for magnified. Nuclei counterstained with 4',6-diamidino-2-phenylindole. Scale Bar: 50 µm; B: Villus height (upper) and crypt depth (lower) of the ileum in the experimental mice; C: Semi-quantitation of ZO-1 and Occludin immuno-positive areas in high-power field by NIH imageJ software; D: Western blots for ZO-1 and Occludin in the liver of experimental mice. Actin was used as internal control; E: Relative mRNA expression levels of Zo1, Ocln, Cldn1 and Cldn4 in the ileum of experimental mice; F: Blood levels of fluorescein isothiocyanate (FITC)-dextran (4kDa) 4 h after oral administration; G and H): Relative mRNA expression levels of Tnfa (G) and Mylk (H) in the ileum of experimental mice. Histochemical quantitative analyses included five fields per section (B and C). The mRNA expression levels were measured by RT-qPCR, and Gapdh was used as internal control (E, G and H). Quantitative values are indicated as fold changes to the values of C/V group (C, E, G and H). Data are mean ± SD (B, C, E, G and H; n = 10, F; n = 5), ^aP < 0.05 and ^bP < 0.01 vs C/V group; ^cP < 0.05 and ^dP < 0.01 vs E/V group; ^eP < 0.05 and ^fP < 0.01 vs E/Zn group; ^gP < 0.05 and ^hP < 0.01 vs E/RFX group. DAPI: 4',6-diamidino-2-phenylindole; ZO-1: Zonula occludens; HE: Hematoxylin and eosin.