Fig. 7.

Inhibition of neuroinflammation by thymoquinone is dependent on SIRT1. Control siRNA- and SIRT1 siRNA-transfected BV2 cells were pre-treated with thymoquinone (10 μM) prior to stimulation with LPS (100 ng/ml) for 24 h. Culture supernatants were analysed for TNFα (a), IL-6 (b), IL-1β (c), nitrite (d) and PGE₂ (e). In f nuclear extracts from cells were added to 96-well plates to which an oligonucleotide containing the NF-κB consensus site (5′-GGGACTTTCC-3′) has been immobilised, followed by addition of NF-κB and HRP-conjugated antibodies. Absorbance was read in a microplate reader. Reduction of cellular ROS production in LPS-stimulated microglia is not dependent on SIRT1 (g). (Mean ± SEM; **p < 0.01, ***p < 0.001 thymoquinone +LPS treatment compared with LPS (alone) in control siRNA-transfected cells; ^# p < 0.05, ^### p < 0.001, thymoquinone + LPS treatment in SIRT1 siRNA-transfected cells compared with thymoquinone + LPS treatment in control siRNA-transfected cells; one-way ANOVA)