Figure 10

Effects of various compounds on the induction of NO in LPS-stimulated thioglycolate-elicited peritoneal macrophages of 8-week-old C57BL/6, BALB/c, double subunits knockout (LMP-7/MECL-1-/-), and PPAR-α, female mice. Thioglycolate-elicited peritoneal macrophages of each mouse were adhered to the bottom of 12 well plates (1 × 10⁷ cells/well in 1.0 mL media) for 4 h. After 4 h, the cells were washed with media three times. The cells were cultured overnight in the fresh media (500 μL) at 37°C in an incubator at 5% CO2 after the final wash. The cells were treated with 100 μL dissolved in 0.2% DMSO of α-tocopherol, 100 μM; δ-tocotrienol, 10 μM; riboflavin, 40 μM; or quercetin-HCL, 40 μM for I h, and the LPS + IFN-γ (10 μL of 1.0 μg/mL + 50 units/mL) was added to assay mixture. The assay mixtures were incubated for 36 h at room temperature. The supernatants were collected after 36 h of LPS challenge, and stored at -70°C to carry out NO estimation, using the Greiss reagent. The cells viability were very good (> 95%) in all the treatments. The treatments 1-5 correspond to: 1. Control (media + macrophages (cells) + LPS + IFN-γ + 0.2% DMSO); 2. α-tocopherol; 3. δ-tocotrienol; 4. riboflavin; 5. quercetin-HCL.