Suppression of nitric oxide induction and pro-inflammatory cytokines by novel proteasome inhibitors in various experimental models

Table 3 Effects of various compounds on the secretion of TNF-α (pg/mL) in LPS-stimulated (pre-treatment) thioglycolate-elicited peritoneal macrophages of 8-week-old BALB/c female mice¹

¹The thioglycolate-elicited peritoneal macrophages (1.0 mL) were adhered to the bottom of 12 well plates (1 × 10⁷ cells/well in 1.0 mL media) for 4 h, then cells were washed extennsively with medium 3-times. Then cells were cultured overnight in fresh media. After overnight incubation at 37°C in an incubator at 5% CO2 for 1 h, then cells were treated with different concentrations of various compounds for 1 h. after 1 h, LPS (10 μL of 1.0 μg/mL stock solution) was added to each well (LPS final concentration 10.0 ng/mL). The supernatants were collected after 4 h of incubation at room temperature. The supernatants were collected after 4 h of LPS challenge, and stored at -70°C to carry out TNF- α ELISA assay. Cells viability were > 95% in all treatments.
²Each compound was dissolved in dimethyl sulfoxide (0.2% DMSO). Appropriate amount of each compound (highest concentration) was dissolved in 1.0 mL DMSO = A stock solution; 50 μL of stock solution A was mixed with 950 μL of media = B working solution. The remaining concentrations were prepared from B working solution (1:2 dilution) successively.
³Percentages of control values are in parenthesis.
*Values from reference [10] in Lipids in Health and Disease 2011,10: 39.

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