Figure 4

Effects of various tocols on secretion of TNF-α in LPS-stimulated RAW 264. 7 cells. RAW 264.7 cells (500 μL) were adhered in wells for 2 h at room temperature. The cells were then treated with various concentrations of α-tocopherol, or α-, γ-, or δ-tocotrienols (100 μL) for 1 h, then the wells were treated with LPS (1 ng/well; 400 μL) for 4 h. The supernatants were then transferred in glass vials and stored at -20°C for subsequent TNF-α assays. Cell viability was determined for all treatment and control groups. The experimental solution were prepared by dissolving highest concentration amount of α-tocopherol or α-, γ-, δ-tocotrienols in 1.0 mL DMSO = X. Solution × (50 μL) was mixed with 950 μL of media = Y. The remaining required concentrations were prepared with Y (1:2 dilutions). The TNF-α assays were carried out using an ELISA kit, and kit control value varies 266 - 444 (287), A = values; B = percentages.