DA-9601, a standardized extract of Artemisia asiatica, blocks TNF-α-induced IL-8 and CCL20 production by inhibiting p38 kinase and NF-κB pathways in human gastric epithelial cells

Figure 4 DA-9601 blocks NF-κB activity in AGS cells. A: AGS cells (1 x 10⁵) were transfected with NF-κB luciferase reporter vector (0.8 μg/well). After 24 h of incubation, cells (1 x 10⁵) were pre-treated for 1 h with DA-9601 (50 μg/mL) and stimulated for additional 16 h with medium alone or medium containing TNF-α (5 ng/mL). At the end of incubation, cells were lysed, and the relative luciferase activity was measured using Luciferase Assay System; B: AGS cells (1 x 10⁵) were transfected with p65-EGFP vector (0.8 μg/well). After 24 h of incubation, cells were pre-treated for 1 h with DA-9601 (50 μg/mL) and stimulated for 1 h with medium alone or TNF-α. Nuclear translocation of p65-EGFP was observed under the fluorescence microscope (original magnification, 200 X); C: AGS cells (5 x 10⁵) were incubated with TNF-α (5 ng/mL) for the indicated time points (0-240 min) (top) or were pretreated with medium alone or with DA-9601 (50 μg/mL) for 1 h, and incubated with TNF-α for 30 min (bottom). The cell lysates were blotted with antibodies specific for the I-κBα and β-actin.