Andrographolide, a Novel NF-κB Inhibitor, Induces Vascular Smooth Muscle Cell Apoptosis via a Ceramide-p47phox-ROS Signaling Cascade

Figure 4

Effects of ROS scavengers on andrographolide-stimulated p53 activation, Bax, and active caspase-3 expression in rat VSMCs. (a) Cells were transiently transfected with PG-13-luc and Renilla-luc for 48 h. After transfection, the cells were pretreated with a vehicle, 1 mM NAC, or 10 𝜇M DPI for 30 min before being treated with 50 𝜇M andrographolide for another 24 h. A PG13-luciferase assay was subsequently conducted. Cells were pretreated with a vehicle, 1 mM NAC, or 10 𝜇M DPI for 30 min before being treated with 50 𝜇M andrographolide for 48 hr, and the expression of Bax (b) and active caspase-3 (c) was examined using immunoblotting. The data are presented as the mean ± SEM (error bars: **𝑃<0.01 and ***𝑃<0.001, compared with the control group, and ^#𝑃<0.05, ^##𝑃<0.01, and ^###𝑃<0.001, compared with the group treated only with andrographolide).

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