NF-B/AP-1-Targeted Inhibition of Macrophage-Mediated Inflammatory Responses by Depigmenting Compound AP736 Derived from Natural 1,3-Diphenylpropane Skeleton

Figure 2

Effects of AP736 on macrophage-mediated inflammatory responses. ((a), (b), and (c)) The effects of AP736 or control compounds (L-NAME and indomethacin) on the levels of NO and PGE₂ were determined in RAW264.7 cells (1 × 10⁶ cells/mL). Cells were treated with LPS (1 μg/mL) in the presence or absence of AP736 for 24 or 6 h. After stimulation, culture supernatants were collected and the resultant concentrations of NO and PGE₂ were determined using the Griess assay and EIAs (left panel). AP736 was incubated with SNP (10 μM) for 30 min in microtubes. The levels of SNP-generated NO in microtubes were also determined by the Griess assay (right panel). (d) RAW264.7 cells (1 × 10⁶ cells/mL) were treated with AP736 for either 8 (left panel) or 24 (right panel) hours. Cell viability was evaluated using the MTT assay. (e) Images of the cells in culture at 6 h were obtained using an inverted phase contrast microscope equipped with a video camera and captured using NIH image software (left panel). Morphologically altered cells were enumerated with a cell counter (right panel). (f) RAW264.7 cells preincubated with AP736 were treated with FITC-dextran (1 mg/mL) for 30 min. The level of dextran uptake was determined by flow cytometric analysis. and compared with the normal or control groups.

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