Fig. 3
TSG inhibited microglia-mediated neuroinflammation. Rat primary midbrain neuron-glia cultures were treated with TSG (20–80 μM) for 30 min before LPS (10 ng/ml) stimulation. Seven days later, LPS-induced microglial activation was analyzed by immunostaining with an anti-OX-42 antibody (a) and Iba-1 protein expression by western blotting (b); the two-way ANOVA interactions analysis showed F = 3.963 and P = 0.127). Images presented were representative of three independent experiments. Scale bar = 200 μm. Also, the total cell protein was harvested and the Iba-1 protein expression was quantified. The ratio of densitometry values of Iba-1 and β-actin was assessed and normalized to the control group. In addition, rat primary midbrain neuron-glia cultures were treated with TSG (20–80 μM) for 30 min and then stimulated by LPS (10 ng/ml). After LPS treatment for 6 and 24 h, an aliquot of the culture medium was harvested for ELISA analysis of TNFα, IL-1β, and PGE2 and Griess reaction analysis of nitrite levels (c). TNFα: the two-way ANOVA interactions analysis of showed F = 3.098 and P = 0.112; NO: the two-way ANOVA interactions analysis of showed F = 4.005 and P = 0.091; PGE2: the two-way ANOVA interactions analysis of showed F = 4.453 and P = 0.087; IL-1β: the two-way ANOVA interactions analysis of showed F = 4.786 and P = 0.063). Data the mean ± SEM from three independent experiments performed in triplicate. #P < 0.05 compared with control cultures; *P < 0.05 compared with LPS-treated cultures