Effect of different BV samples and BV-4 on inflammatory mediator expression.

<p>In (A), RAW 264.7 macrophages (1×10⁶ in 2 ml of medium) were incubated for 30 min with or without the indicated concentrations of BV-1, BV-2, BV-3, or BV-4, then for 24 h with or without addition of 1 µg/ml of LPS, then NO generation in the culture medium was measured by the Griess reaction. In (B), (C), and (D), RAW 264.7 macrophages (1×10⁶ in 2 ml of medium) were incubated for 30 min with or without the BV-4, then for 24 h with or without addition of 1 µg/ml of LPS, then IL-6 (B) and TNF-α (C) in the culture medium were measured by ELISA and levels of iNOS and COX-2 (D) in cell lysates were measured by Western blotting. In (E), RAW 264.7 macrophages (5×10⁴ in 1 ml of medium) were incubated for 30 min with or without the indicated concentration of BV-4 or 50 µM cinnamaldehyde, then for 24 h with or without addition of 1 µg/ml of LPS, then cell viability was measured using the AlamarBlue® assay. In (A), (B), (C), and (E), the data are expressed as the mean ± SD for three separate experiments, while, in (D), the lower panel shows a typical result and the histogram shows results for 3 experiments expressed as the mean ± SD. # indicate a significant difference at the respective level of <i>p</i><0.001 compared to the LPS-treated group.