Fig. 4

Ethanol-mediated potentiation of LPS-induced inflammatory responses and ameliorating effect of synaptamide in microglia and peritoneal macrophages in culture. Microglia and peritoneal macrophages were isolated from adult mice, incubated with 25 mM ethanol for 4 h and treated with LPS (100 ng/ml) for 1 or 12 h for mRNA or protein analysis, respectively. Synaptamide (10 nM) was added to the cell culture immediately after LPS treatment. The mRNA expression of IL-1β, TNF-α or IL-6 and TNF-α protein level were measured in microglia (A) and peritoneal macrophages (B) and the data are presented as the fold change relative to the maltose control (MAL). Ethanol potentiated the LPS-induced proinflammatory cytokine expression at both mRNA and protein levels while synaptamide suppressed the effect of LPS and ethanol in microglia and macrophage cells in culture. Values are presented as mean ± SEM (n = 3–4), representing two independent experiments. ns, the difference of means is not statistically significant. *p < 0.05; **p < 0.01; ***, p < 0.001; ****p < 0.0001 vs. Maltose group