Figure 6.

2′MeO6MF decreases NFkB activity and circulating pro-inflammatory cytokines. The level of expression for various GABA subunits was assessed in the RAWblue™ macrophage cell line. Macrophage cells were shown to express α5, β1 and δ, with very low levels of α1, α2 and β2 being detected (a). Upon stimulation of macrophage with LPS (2.5 ng/ml) a significant increase in NFkB activity was detected (b). Addition of the 2′MeO6MF (10⁻³–10⁻⁴), but not the vehicle (2.5% DMSO), to the media resulted in a significant decrease in NFkB activity (b). Data are presented as mean ± SD. +++=P < 0.001 compared with controls; ^=P < 0.01 compared to LPS or LPS + DMSO-treated controls. 2′MeO6MF: 2′-methoxy-6-methylflavone. The effects of sham-operated controls (white bars), stroke + vehicle treatment (blue bars) and stroke + 2′MeO6MF treatment (red bars) on circulating IL-1β (c), TNF-α (d) and IFN-γ levels (e) were assessed in plasma collected 1 - (solid bars) and three days (chequered bars) post-stroke. Stroke induces a significant elevation in circulating pro-inflammatory IL-1β (c), TNF-α (d) and IFN-γ (e) levels. Treatment with 2′MeO6MF dosing at 1 h with a second dose at 24 h resulted in a significant decrease in circulating IL-1β and IFN-γ ((c) and (e), respectively) levels at 0.1, 5 and 30 mg/kg. Treatment with 2′MeO6MF also resulted in a significant decrease in circulating TNF-α (d) levels three days following dosing at 30 mg/kg, but not on day 1, nor following dosing at the either 0.1 or 5 mg/kg. All data are presented as mean ± SD. +=P < 0.05, ++=P < 0.01 and +++=P < 0.001 compared with sham-operated controls; *=P<0.05, ^=P < 0.01 compared with stroke + vehicle-treated controls. DMSO: ; IF: interferon; LPS: ; TNF: tumour necrosis factor.