Fig. 4

ACT001 mitigated LPS-induced pro-inflammatory activation of BV2 cells in vitro. a BV2 cells were treated with indicated doses of ACT001 for 12–48 h, then the cytotoxicity of ACT001 was measured by CCK-8 assay. The cell viability result was normalized to BV2 cells without ACT001 treatment (Control) for 12 h. b, c Indicated doses of ACT001 were co-treated with 100 ng/ml LPS (b) or 500 ng/ml LPS (c) in BV2 cells for 12–48 h, then the cytotoxicity of co-treatment was measured by CCK-8 assay. The cell viability result was normalized to BV2 cells without ACT001 and LPS treatment (Control) for 12 h. d The effect of ACT001 and LPS co-treatment on morphological changes of BV2 cells. The normal morphology of BV2 cells cultured in complete DMEM for 24 h was shown as Control. Red arrows indicated inflammatory amoeboid morphology of activated BV2 cells. Scale bar = 50 μm. e, f After co-treatment with indicated doses of ACT001 and 500 ng/ml LPS for 24 and 48 h, relative mRNA expression levels of pro-inflammatory cytokines (e) and anti-inflammatory cytokines (f) in BV2 cells were quantified by Real-time PCR. Cells without ACT001 and LPS treatment were shown as Control. Data were presented as means ± SEMs of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 versus Control group; # P < 0.05, ## P < 0.01, ### P < 0.001 versus ACT001 0 μM + LPS group