Inhibition of microglial activation by PC(16:0/16:0). Cultured mouse brain primary microglia were treated with PC(16:0/16:0) (A) and LPC(16:0) (B) at the indicated concentrations. Cellular toxicity was assessed using the CCK-8 viability kit. C, micrographs showing primary microglia untreated (left panel) and LPS-treated (right panel), followed by immunohistochemical staining with Iba1 antibody. The scale bar = 20 μm. ELISA assay was performed to show LPS-induced significant expression of IL-1β in cultured microglia (D), but not by LPC(16:0) and PC(16:0/16:0) treatments. Adding LPS with PC(16:0/16:0) at the indicated dosage significantly inhibited LPS-induced IL-1β expression (E), but not with LPC(16:0) (G). Similarly, PC(16:0/16:0) also significantly inhibited LPS-induced induction of TNK-α, but not by LPC(16:0). Data represent the mean ± SD. Error bars indicate SD. n = 5 independent repeats; ∗ indicates p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 by one-way ANOVA with LSD post hoc analysis to identify significant groups.