Figure 6.

Effects of EEP on LPS-induced production of NO and PGE₂ and expression of iNOS and COX-2 in RAW 264.7 macrophages. Following treatment with or without indicated dose of EEP for 1 h, cells were treated with or without LPS (1 μg/mL) for 24 h. Control cells were not treated with EEP or LPS. (A,B) Production of NO and PGE₂ was determined by Griess reaction and EIA kits, respectively. l-NIL (40 µM) and NS-398 (10 nM) were used as positive control for NO and PGE₂ production, respectively. (C) Total cellular proteins were obtained from cells stimulated with LPS (1 μg/mL) for 24 h with or without EEP (50, 100, or 200 μg/mL). Protein expression levels of iNOS and COX-2 were detected by Western blotting. β-Actin was used as an internal control. (D,E) Total RNAs were prepared from cells stimulated with LPS for 6 h in presence or absence of EEP (50, 100, or 200 μg/mL). mRNA levels of iNOS and COX-2 were determined by qRT-PCR using specific primers. Results were normalized against β-actin. Experiments were performed independently three times and similar results were obtained. Values shown are the means ± SDs of three independent experiments. # p < 0.05 vs. control group; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. LPS-stimulated group.