Inhibitory Effects of Traditional Herbal Formula Pyungwi-San on Inflammatory Response In Vitro and In Vivo
Inhibitory effects of PWS on LPS-induced production of PGE2 (a), NO (b), and pro-inflammatory cytokines (IL-6 (c) and TNF-α (d)) in Raw 264.7 macrophages. Cells (5 × 105 cells/mL) were treated with various concentrations (0.5, 0.75, and 1 mg/mL) of PWS for 1 h, followed by continuous incubation with LPS (1 μg/mL) for the next 20 h. Concentrations of PGE2, NO, and pro-inflammatory cytokines in culture medium were monitored as described in the Materials and Methods Section. The MTT assay was used for measurement of viability of cells exposed to PWS (e). Inhibitory effects of PWS on LPS-induced NF-κB dependent luciferase activity (f). Transfected cells (5 × 105 cells/mL) were incubated for 16 h and pretreated with different concentrations (0.5, 0.75, and 1 mg/mL) of PWS for 1 h, followed by stimulation with 1 μg/mL LPS for 20 h. Following lysis of cells, measurement of luciferase activity was performed using the Promega luciferase assay system and a luminometer. Data represent the mean ± S.D. from three separate experiments. **, ***, significant compared with vehicle-treated control; #, ##, ###, significant compared with LPS alone.