Fig. 4

Zafirlukast effectively inhibits the LPS-induced inflammation of epithelial cells through TLR4/NF-KB/NLRP3 pathway in vitro. A-B A549 cells were subjected to different concentration of LPS or Zafirlukast (2.5 μM, 5 μM, 10 μM, 20 μM) for a duration of 24 h, Western blotting was then performed for assessing the protein expression levels of MyD88, TLR4, P65, P-P65 as well as inflammasome pathway-related protein NLRP3, caspase1, Cleaved-caspase1, IL-1β and Cleaved-IL-1β in the cells (A). The quantification of optical density was also determined (B). C-D Additionally, the activation of MYD88 (C) and P-P65 (D) by immunofluorescence in A549 cells subjected to LPS or Zafirlukast (2.5 μM, 5 μM, 10 μM, 20 μM) for 24 h (Scale: 50 μm). (E–F) MLE-12 cells were subjected to LPS or Zafirlukast (2.5 μM, 5 μM, 10 μM, 20 μM) for 24 h, and the Western blotting was used to assess the MyD88, TLR4, P65, P-P65 and the inflammasome pathway related protein NLRP3, caspase1, Cleaved-caspase1, IL-1β and Cleaved-IL-1β protein expression levels in the cells (E). The optical density was quantified and presented (F). G-H MLE-12 cells were subjected to LPS or Zafirlukast (2.5 μM, 5 μM, 10 μM, 20 μM) for 24 h, and the activation of MYD88 (G) and P-P65 (H) was assessed through immunofluorescence (Scale: 50 μm). Data are shown as mean ± SD (n = 3). Statistical significance was denoted as follows: #P < 0.05, ##P < 0.01, ###P < 0.001 compared with control group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 compared with model group