Fig 1.
(a) Changes in the expression of claudin-1 (b) and occludin (c) proteins detected by WB. Data are presented as the mean ± SD. four independent experiments were performed in duplicate. *P < 0.05 vs. control group. RT-qPCR detection of TNF-α (d) and IL-6 (e) RNA expression in cells with different concentrations of LPS. Data are presented as the mean ± SD. three independent experiments were performed in duplicate. *P < 0.05 vs. control group, **P < 0.01 vs. control group. ELISA was used to detect the expression of TNF-α (f) and IL-6 (g) when different concentrations of LPS interfered with cells. Data are presented as the mean±SD. three independent experiments were performed in duplicate. *P < 0.05 vs. control group, **P < 0.01 vs. control group.
Fig 2.
Measurement of TEER at 0, 24, and 48 h by each cell resistance meter (a) and TEER changes of each group at 24–48 h (b), Data are presented as the mean ± SD. three independent experiments were performed in duplicate. **P < 0.01 LPS + LP group vs. LPS group. (c) Effect of LPS treatment for 48 h on cell viability. (d) WB detection of claudin-1, JAM-a, occludin, and Zo-1 expression levels. Data are presented as the mean ± SD. four independent experiments were performed in duplicate. *P < 0.05 LPS + LP group vs. LPS group, **P < 0.01 LPS + LP group vs. LPS group. (e) Immunofluorescence detection of ZO-1 tight junction protein expression and distribution. three independent experiments were performed in duplicate. (f) Changes in electron microscopy-observed tightly connected cellular ultrastructure between groups. Arrows indicate the tight connection of Caco2 cells in each group A: Ctrl group, B: LPS group, C: LPS + LP group, D: LP group. three independent experiments were performed in duplicate.
Fig 3.
FSK toxicity on cells at different concentrations (a) in the experiment. Data are presented as the mean ± SD. five independent experiments were performed in duplicate.*P < 0.05 vs. control group. Toxicity of different concentrations of HT89 (b) on cells. Data are presented as the mean ± SD. three independent experiments were performed in duplicate. *P < 0.05 vs. control group, **P < 0.01 vs. control group. (c)-(d) TEER changes in each group at 0, 24, 48, and 24–48 h. Data are presented as the mean ± SD. three independent experiments were performed in duplicate. **P < 0.01 vs. control group or LPS + LP + HT89 group vs. LPS + LP group.
Fig 4.
(a) WB detection of tight junction protein (claudin-1, JAM-a, occludin, and Zo-1) expression. Data are presented as the mean ± SD. three independent experiments were performed in duplicate. *P < 0.05 LPS + LP + HT89 group vs. LPS + LP group or LPS + LP group vs. LPS group. **P < 0.01 LPS + LP + HT89 group vs. LPS + LP group. (b) Immunofluorescent detection of the expression and distribution of the tight junction protein, ZO-1. three independent experiments were performed in duplicate. (c) Electron microscopy-observed changes of the tightly connected ultrastructure of cells between groups. Arrows indicate the tight connection of Caco2 cells in each group. A: Ctrl group, B: LPS group, C: LPS + LP group, D: LPS + FSK group, E: LPS + LP + HT89 group. three independent experiments were performed in duplicate.