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Figure 9 | Journal of Neuroinflammation

Figure 9

From: Inhibitory effect of 4-O-methylhonokiol on lipopolysaccharide-induced neuroinflammation, amyloidogenesis and memory impairment via inhibition of nuclear factor-kappaB in vitro and in vivo models

Figure 9

Effect of 4- O -methylhonokiol on LPS-induced iNOS, COX-2, APP, C99, Aβ, BACE generation, NF-κB activity, β- and γ-secretase activity in cultured astrocytes and in microglial BV-2 cells. Astrocytes and microglial BV-2 cells were treated with LPS (1 μM) and 4-O-methylhonokiol (0.5-2 μM). (A) NF-κB activity in astrocytes was determined by EMSA as described in Methods. (B) For supershift assays, nuclear extracts from cultured astrocytes treated with LPS (1 μg/ml) were incubated for 1 h before EMSA with specific antibodies against the p50 and p65 NF-κB isoforms. (C) Phosphorylation of IκB, and p50 and p65 translocation in the astrocytes. (D) Immunoblots of lysates from astrocytes were probed with iNOS, COX-2, APP, C99, Aβ and BACE antibodies, respectively. (E) The amounts of Aβ1-42 were assessed by using a specific Aβ1-42 ELISA kit as described in the Methods. (F) and (G) β- and γ-secretase activity in the astrocytes were determined as described in Methods. (H) Immunoblots of lysates from microglial BV-2 cells were probed with iNOS, COX-2, APP, C99 and BACE antibodies, respectively. (I) The amounts of Aβ1-42 were assessed by using a specific Aβ1-42 ELISA kit as described in the Methods. Values represent means ± S.D. of three independent experiments performed in triplicate. The values in the western blot band indicate average density over β-actin from three animals. #, Significantly different from control group (p < 0.05). *, Significantly different from LPS-treated group (p < 0.05). Control, saline-treated group. LPS, lipopolysaccharide. MH, 4-O-methylhonokiol.

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