Toona sinensis Inhibits LPS-Induced Inflammation and Migration in Vascular Smooth Muscle Cells via Suppression of Reactive Oxygen Species and NF-B Signaling Pathway
Figure 8
TS and GA suppress LPS-induced NF-κB transcriptional activation and IκB degradation in A7r5 cells. (a) A7r5 cells were transiently transfected with SEAP plasmids using lipofectamine. NF-κB transcriptional activation was measured after preincubation with or without TS (25–100 μg/mL) or GA (5 μg/mL) for 2 h, followed by stimulation with LPS (100 ng/mL) for 3, 6, 9, and 12 h. Cell lysates were mixed with luciferase reagents and quantified using a illuminometer. Relative NF-κB activity was calculated by dividing the relative luciferase unit (RLU) of treated cells by the RLU of untreated cells. (b) Western blotting was performed to analyze IκB protein levels. Cells were preincubated with or without TS (25–100 μg/mL) or GA (5 μg/mL) for 2 h and then stimulated by LPS (100 ng/mL) for 45 min. β-Actin was used as an internal control. Relative changes in protein bands were measured using AlpaEaseFc 4.0 software. Densitometric analysis, with the control being 100%, is shown below the gel data. The results are presented as the mean ± SD of three assays. significant difference () in comparison to control or LPS-treated groups, respectively.
