Evidence-Based Complementary and Alternative Medicine / 2018 / Article / Fig 4

Research Article

Anti-Inflammatory Potential of Carpomitra costata Ethanolic Extracts via Inhibition of NF-κB and AP-1 Activation in LPS-Stimulated RAW264.7 Macrophages

Figure 4

Effects of CCE on LPS-induced phosphorylation of MAPKs and Akt in RAW264.7 macrophages. RAW264.7 cells were pretreated with either the vehicle or the indicated concentration of CCE (30–100 μg/mL) for 30 min, prior to stimulation with LPS (100 ng/mL). (a) Cell extracts were then prepared and subjected to western blotting with antibodies specific for the total and phosphorylated forms of MAPKs (p38, ERK1/2, JNK) (for 15 min) and Akt (for 4 h). The results are representative of three independent experiments. Levels of NO and PGE2 in the supernatant were detected using the Griess reaction assay (b) and ELISA (c). (d) The expression levels of TNF-α, IL-6, and IL-1β in the culture medium were determined using ELISA. Cells were treated with SP600125 (a JNK inhibitor) and LY294002 (an Akt inhibitor) 30 min prior to LPS stimulation for 24 h. Each value indicates the mean ± SEM and is representative of the results obtained from three independent experiments. versus control group; versus LPS-stimulated group. SP: SP600125 and LY: LY294002.