Fig. 1. Effects of BST204 on COX-2 expression in RAW 264.7 macrophages. (A) The expression of COX-2 was time-dependently induced by LPS in RAW 264.7 macrophages. The cells were incubated with 1-μg/ml LPS for the indicated times and the levels of COX-2 mRNA and protein were detected by RT-PCR and Western blot analysis, as described in Materials and methods. (B) Effects of BST204 on the induction of COX-2 expression. The cells were incubated with 20-μg/ml BST204 for 4, 8, or 16 h or with crude ginseng extract (GE, 50 μg/ml) for 16 h and the levels of COX-2 protein were detected by Western blot analysis.

Fig. 2. BST204 inhibited the expression of COX-2 protein, but not the level of COX-2 mRNA. (A) The cells were treated with BST204 in the indicated doses or with crude ginseng extract (GE, 50 μg/ml) 1 h before they were incubated with 1-μg/ml LPS for 4 h, and the levels of COX-2 protein were detected by Western blot analysis. (B) The PGE₂ production was dose-dependently inhibited by BST204, not by crude ginseng extract (GE). The cells were treated with BST204 in the indicated doses or with crude ginseng extract (GE, 50 μg/ml) 1 h before they were incubated with 1-μg/ml LPS for 16 h, and the levels of PGE₂ in media were detected using a specific enzyme immunoassay according to the manufacturer’s instruction (Cayman Co., MI). (C) The cells were treated with BST204 in the indicated doses or with crude ginseng extract (GE, 50 μg/ml) 1 h before they were stimulated with 1-μg/ml LPS for 4 h and the levels of COX-2 mRNA were measured by RT-PCR, as described in Materials and methods.

Fig. 3. The activating phosphorylation of p70 S6 kinase was dose-dependently inhibited by BST204 but not by crude ginseng extract. (A) The cells were treated with 1-μg/ml LPS for the indicated times and the phosphorylation of p70 S6 kinase was detected by Western blot analysis. (B) The cells were treated with BST204 in the indicated doses or with crude ginseng extract (GE, 50 μg/ml) 1 h before they were incubated with 1-μg/ml LPS for 4 h and the phosphorylation of p70 S6 kinase was detected by Western blot analysis, as described in Materials and methods.

Fig. 4. Effects of rapamycin on the expression of COX-2 protein in RAW 264.7 macrophages. The cells were treated with rapamycin in the indicated doses 1 h before they were incubated with 1-μg/ml LPS for 4 h, and the levels of COX-2 protein and the phosphorylation of p70 S6 kinase were detected by Western blot analysis, as described in Materials and methods.

Ginsenoside contents of the fermented ginseng extract compared to crude ginseng extract

The extracts were solubilized in methanol and for the analysis of ginsenosides, high performance liquid chromatography was accomplished as in Materials and methods.