Inhibition of JNK and p38 in macrophages suppressed inflammatory cytokine expression in macrophages and in macrophage-adipocyte interaction.

<p><i>(A)</i> RAW264.7 cells were stimulated with IFN-γ plus LPS in the presence of JNK-specific inhibitor SP600125 (20μM) (Sigma) or p38-specific inhibitor SB23580 (20μM) for the indicated time points. DMSO was used as vehicle control. The expression and activation of JNK or ATF2 were determined by western blot analysis. RAW264.7 cells were pretreated with SP600125 (20μM) followed by stimulation with M1 <i>(B)</i> or M2 <i>(C)</i> activation condition. The expression of M1 or M2 markers was determined. RAW264.7 cells were pretreated with SB23580 (20μM) followed by stimulation with M1 <i>(D)</i> or M2 <i>(E)</i> activation condition. The expression of M1 or M2 genes was analyzed. <i>(F)</i> Control and MKP-2 overexpressing RAW264.7 cells were pretreated with SP600125 (20μM) or SB23580 (20μM) or both for 1h before co-culture with differentiated 3T3-L1 adipocytes for 24h. The concentrations of MCP-1, IL-6, and TNF-α in culture supernatants were determined by ELISA. Cytokines production was normalized to total protein of the cell lysates. Ct, control; Co, co-culture. Data shown are representative of three independent experiments with similar results. Data are presented as mean ± SEM. *, p<0.05; **, p<0.01; ***, p<0.001.