Figure 11.

ST1926 suppresses inflammation response and reactive oxygen species (ROS) production in lung epithelia cells in vitro. (A) Immunofluorescence of phosphorylated nuclear factor-κB (NF-κB) was evaluated. (B) Tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-18 and IL-6 mRNA levels were calculated via RT-qPCR analysis. (C) Western blot analysis was used determine Toll-like receptor 4 (TLR4) and MyD88 expression levels in lipopolysaccharide (LPS)-induced cells with or without ST1926. (D) Inhibitor-κB kinase (IKKα) and inhibitor-κB kinase-α (IκBα) protein expression levels were calculated through western blot analysis. (E) ROS levels were calculated in different groups of cells. (F) SOD1 and SOD2 mRNA levels were determined via RT-qPCR analysis. (G) Phosphorylated p38, and (H) extracellular receptor kinase 1/2 (ERK1/2) was determined by the use of western blot analysis. (I) MLE-12 cells were treated with 100 ng/ml LPS for different times as indicated. Then, western blot analysis was conducted to investigate p38 and ERK1/2 phosphorylation. (J) ST1926 reduced p38 and ERK1/2 phosphorylation in MLE-12 cells after LPS treatment in the presence or absence of ST1926 for different times from 0 to 48 h. Western blot assays were used to explore p38 and ERK1/2 activation. (K) Western blotting was carried out to investigate nuclear NF-κB levels. (L) Cytoplasm p-IκBα and IκBα levels were determined by western blot analysis. The ratio of p-IκBα to IκBα was exhibited. The data are represented as mean ± SD (n=8). ^*p<0.05 and ^**p<0.001 vs. the control (Con); ⁺p<0.05, ⁺⁺p<0.01 and ⁺⁺⁺p<0.001 vs. LPS-induced mice (LPS).