Figure 8

Effects of various compounds on the productions of nitric oxide (NO) in LPS-stimulated (pre-treatment) peritoneal macrophages of 8-week-old C57BL/6 versus BALB/c female mice. Thioglycolate-elicited peritoneal macrophages were prepared from 8-week-old C57BL/6 and BALB/c (5 of each) female mice as described previously [6]. The macrophages (1 x10⁷ cells/well in 1.0 mL media) were adhered to the bottom of 100 mm tissue culture plate for 4 h. After 4 h the cells were treated with each compound (100 μL dissolved in 0.2% DMSO) of dexamethasone, 10 μM; mevinolin, 20 μM; α-tocopherol, 100 μM; δ-tocotrienol, 10 μM; riboflavin, 40 μM; or quercetin-HCL, 40 μM for I h, followed by stimulation with LPS 10 n g/mL (10 μL of 1.0 μg/mL) of each treatment. The assay mixtures were incubated at room temperature for 4 h, after 4 h, the assay mixtures were centrifuged at 2,000 rpm for 20 min. The cells were then harvested, and the total cellular RNA was extracted from each pellet with RNeasy mini kit according to manufacturer's instructions [17]. The total level of NO for each inhibitor was estimated in the supernatant by using Griess reagent as described earlier [7, 17]. The cells viability were very good (> 95%) in all the treatments [4, 7]. Data are means ± SD, n = 5 per treatment, and triplicate analyses of each sample. The treatments 1-7 correspond to: 1. Control (macrophages + LPS + 0.2% DMSO); 2. dexamethasone; 3. mevinolin; 4. α-tocopherol; 5. δ-tocotrienol; 6. riboflavin; 7. quercetin-HCL. Values in a column with a different superscript symbols are significantly different at P< 0.05, A = values, B = Percentages.