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

Figure 6 Effects of zinc acetate and rifaximin on in vitro EtOH/LPS/TNF-α-stimulated Caco-2 cells. A-D: In vitro paracellular permeability in ethanol (EtOH) (A and B)- or lipopolysaccharide (LPS) (C and D)-stimulated Caco-2 cells determined as transepithelial electrical resistance; E and F: Western blots for the effects of zinc acetate (100 μM) on ZO-1, Occludin, p-AKT and AKT expressions (E) and rifaximin (10 μM) on ZO-1, Occludin, p-p65 and p65 expressions (F) in the whole cell lysate of Caco-2 cells. Actin was used as internal control; G and H: Relative mRNA expression levels of MYLK in TNF-α-stimulated Caco-2 cells. The mRNA expression levels were measured by RT-qPCR, and GAPDH was used as internal control. Quantitative values are indicated as fold changes to the values of non-treatment group. Caco-2 were treated with each agent as following; (A, C, E and G) zinc acetate (Zn) and/or PI3K inhibitor, LY294002, (B, D, F and H) rifaximin (RFX) and/or human PXR inhibitor, SPA70. Data are mean ± SD (A-D; n = 6, G and H; n = 8), ^aP < 0.01 vs non-treated groups (A-D, G and H), ^bP < 0.01 vs EtOH (A and B) or LPS-treated groups (C and D), ^cP < 0.01 vs EtOH with Zn (100 μM) (A), EtOH with RFX (10 μM) (B), LPS with Zn (100 μM) (C) or LPS with RFX (10 μM) (D)-treated groups, ^dP < 0.01 vs TNF-α-treated group (H), ^eP < 0.01 vs TNF-α with RFX (10 μM)-treated group (H). LPS: Lipopolysaccharide; TEER: Transepithelial electrical resistance; EtOH: Ethanol; ZO-1: Zonula occludens.