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
| Pre-treatment | Secretion of TNF-α, (pg/mL) | ||||||
|---|---|---|---|---|---|---|---|
| # | Assay Mixture | Dexamethasone | Mevinolin | α,-Tocopherol | δ-Tocotrienol* | Riboflavin | Quercetin |
| 1 | Media + Cells = A | ||||||
| 2 | A + LPS (10 ng/mL) = B | 1224 | 1224 | 1224 | 1224 | 1224 | 1224 |
| 3 | B + 0.2% DMSO2 = C | 1305 (100)3 | 1305 (100)3 | 1305 (100)3 | 1305 (100)3 | 1305 (100)3 | 1305 (100)3 |
| 4 | C + 5 μM | 333 (26) | 1125 (86) | 1280 (98) | 938 (72) | 1012 (76) | 980 (75) |
| 5 | C + 10 μM | 283 (22) | 1064 (82) | 1267 (97) | 962 (74) | 896 (69) | 873 (67) |
| 6 | C + 20 μM | 234 (18) | 1012 (78) | 1235 (95) | 834 (64) | 674 (52) | 846 (65) |
| 7 | C + 40 μM | 213 (16) | 1048 (80) | 1259 (96) | 769 (59) | 621 (48) | 821 (63) |
| 8 | C + 80 μM | 207 (16) | 1140 (87) | 1298 (99) | 1673 (128) | 544 (42) | 778 (60) |
| 9 | C + 160 μM | 133 (10) | 1396 (107) | 1323 (101) | 1732 (133) | 520 (40) | 472 (36) |
| 10 | C + 320 μM | 112 (9) | 1443 (111) | 1257 (96) | 1987 (152) | 480 (37) | 345 (26) |
| 11 | C + 640 μM | 85 (7) | 1487 (114) | 1241 (95) | 2245 (172) | 406 (31) | 310 (24) |
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.