Figure 4.

FA protected against LPS-induced intestinal epithelial barrier dysfunction by up-regulation of miR-200c-3p. (A) Caco-2 cells were transfected with Lv-miR-200c-3p (Lv-200c), Lv-miR-200c-3p spong (Lv-200c spong), and Lv-NC at a MOI of 15 and incubated at 37°C with 5% CO₂ for 48 h, the expression levels of miR-200c-3p were measured by qRT-PCR. (B, C) Cells were transfected with Lv-200c, Lv-200c spong or Lv-NC at an MOI of 15 and incubated at 37°C with 5% CO₂ for 48 h, and then cells were pretreated with FA (100 μM) for 2 h and stimulated with LPS for 24 h. TER values were monitored across the cell monolayers using Millicell-ERS and permeability of FD4 across the cell monolayer was measured. (D) The protein expression levels of occludin and ZO-1 were determined by Western blot analysis. Data were presented as means ± SD from three independent experiments and differences between means were compared using one-way ANOVA with Tukey’s multiple comparisons test. *P < 0.05, **P < 0.01. (E) The distribution and expression of ZO-1 were detected by immunofluorescence assay. ZO-1 (green) was labeled with fluorescent secondary antibodies and nuclei (blue) were labeled with DAPI (Scale bar = 50 μm).