Figure 7.

DHA supplementation suppresses nigericin-induced IL-1β and IL-1α release and cell death in RAW-ASC MΦs. RAW-ASC cells were incubated in serum-deprived RPMI containing DHA (0, 10, or 25 μM) or VEH (BSA) for 24 h. Cells were pretreated with 20 ng/ml LPS for 2 h, incubated with 0, 2.5, or 5.0 μM nigericin for 45 min, and then release of (A) IL-1β, (B) IL-1α, and (C) LDH measured. The presence of mature (D) IL-1β and (E) IL-1α in the supernatant were determined by Western blotting. For Western blots of IL-1α, LPS priming was extended to 5 h and supernatant concentrated 10x by methanol-chloroform precipitation. Data presented as mean ± SEM, n = 3. Significant differences between DHA-supplemented groups represented as different letters (uppercase for 5 μM group, lowercase for 2.5 μM group). Significant differences from vehicle control at each DHA concentration represented by asterisks (*p < 0.05, **p < 0.01). ELISAs and LDH assay are representative of three independent experiments. Western blots are representative of two independent experiments.