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Table 5 δ-Tocotrienol and riboflavin inhibit expression of TNF-α, IL-1β, IL-6, and iNOS genes in LPS-stimulate RAW 264.7 cells, and peritoneal macrophages derived from C57BL/6 or BALB/c mice 1

From: Inhibition of nitric oxide and inflammatory cytokines in LPS-stimulated murine macrophages by resveratrol, a potent proteasome inhibitor

NO Treatments RT-PCR data (Ratios of digital values of optical density of gene expression of cytokines/β-actin).
TNF-α IL-1β IL-6 iNOS
RAW 264.7 cells.
1 Media + Cells = A 0.05 0.05 0.1 0.2
2 A + LPS (10 ng/well) = B 0.83 1.35 0.98 1.12
3 B + 0.2 % DMSO = C 0.80 ± 0.03a (100)2 1.32 ± 0.03a (100)2 0.96 ± 0.03a (100)2 1.04 ± 0.02a (100)2
4 C + δ-Tocotrienol (16.0 μM) 0.32 ± 0.03c (40) 0.33 ± 0.03c (25) 0.41 ± 0.04c (43) 0.42 ± 0.04c (40)
5 C + Riboflavin (16.0 μM) 0.64 ± 0.04b (80) 0.95 ± 0.03b (72) 0.82 ± 0.04b (85) 0.76 ± 0.03b (73)
8-week-old C57BL/6
1 Media + Cells (macrophages) = A 0 0 0 0
2 A + LPS (10 ng/well) = B 0.35 5.32 11.67 4.97
3 B + 0.4 % DMSO = C 0.36 ± 0.02a (100)2 6.89 ± 0.86a (100)2 12.34 ± 1.1a (100)2 5.67 ± 0.67a (100)2
4 C + δ-Tocotrienol (40.0 μM) 0.16 ± 0.08c (45) 2.34 ± 0.42c (34) 1.98 ± 0.24c (16) 2.12 ± 0.56c (56)
5 C + Riboflavin (40.0 μM) 0.22 ± 0.02b (61) 4.15 ± 0.68b (60) 7.23 ± 0.98b (59) 4.16 ± 0.53b (73)
8-week-old BALB/c
1 Media + Cells (macrophages) = A 0 0 0 0
2 A + LPS (10 ng/well) = B 0.62 8.27 14.45 7.76
3 B + 0.4 % DMSO = C 0.74 ± 0.05a (100)2 10.67 ± 1.24a (100)2 15.67 ± 1.45a (100)2 9.85 ± 0.88a (100)2
4 C + δ-Tocotrienol (40.0 μM) 0.39 ± 0.01c (53) 4.16 ± 0.36c (39) 3.45 ± 0.28c (22) 6.64 ± 0.86b (67)
5 C + Riboflavin (40.0 μM) 0.54 ± 0.03b (73) 6.45 ± 0.91b (60) 4.73 ± 0.22b (30) 7.64 ± 0.67b (78)
  1. 1RAW 264.7 cells or thioglycolate-elicited peritoneal macrophages from 8-week-old C57BL/6 or BALB/c mice were treated with δ-tocotrienol or riboflavin (16 μM or 40 μM) for 1 h, followed by treatment with LPS (10 ng/well) for 4 h. Total RNA was extracted, reverse transcribed, and the resulting DNA was amplified and analyzed by real time PCR to quantitate expression of TNF-α, IL-1β, IL-6, and iNOS genes [5, 21].
  2. Cell viability was >95 % in all the treatments. Data are means ± SD, n = 3 (triplicate analysis of each sample).
  3. 2Percentages of digital values of relative optical density of each cytokine/β-actin of each treatment are in parenthesis. a-cValues in a column not sharing a common superscript letter are significantly different at P < 0.05.