Ethyl Acetate Extract of Artemisia anomala S. Moore Displays Potent Anti-Inflammatory Effect
Figure 5
Effect of EAFA on proinflammatory cytokine and HO-1 expression in LPS/IFN-stimulated RAW264.7 cells. (a, b) RAW264.7 cells ( cells/dish) were pretreated with varying doses of EAFA for 1 h and then stimulated with IFN (10 U/mL) plus LPS (100 ng/mL) for 6 h. Total RNA was isolated and subjected to qRT-PCR to determine the level of IL-1 (a) and IL-6 mRNA (b). (c, d) RAW264.7 cells were treated with IFN (10 U/mL) plus LPS (100 ng/mL) in the presence of varying concentrations of EAFA for 24 h. Conditioned media were collected and subjected to ELISA to determine the amount of IL-1 (c) and IL-6 (d). The values (means SEM) were obtained from three independent experiments. ##, # versus control group; **, * versus model group. (e) RAW264.7 cells ( cells/dish) were pretreated with varying concentrations of EAFA (50, 100, 200 g/mL) for 1 h followed by LPS (100 ng/mL) plus IFN (10 U/mL) treatment for 18 h. Total RNA was isolated and subjected to qRT-PCR to measure HO-1 mRNA level. -actin was used as an internal control for standardization. (f) RAW264.7 cells plated at a density of cells/well in 30 mm dish for overnight were pretreated with EAFA for 1 h followed by 18 h-stimulation of IFN (10 U/mL) plus LPS (100 ng/mL). Whole cell lysates were prepared and subjected to western blotting to determine HO-1 protein levels. Data are the representative of three independent experiments. ##, # versus control group; *, ** versus model group.