Fig. 5.

TLR4/MyD88/NF-κB signaling pathway participated in free heme-induced neuroinflammation and cognitive dysfunction. A Representative western blot bands of TLR4, MyD88, p-P65 and P65 in rat brain tissue on day 7 after administration. B Relative intensity of TLR4, MyD88 and p-P65 protein expression in rat brain tissue on day 7 after administration. C Representative western blot bands of TLR4, MyD88, p-P65 and P65 in BV2 cells after treatment for 24 h. BV2 cells were divided into 4 groups: 1) the Control group; 2) the Heme group treated with hemin; 3) the Heme + T6167923 group treated with hemin and MyD88 inhibitor; and 4) the Heme + JSH-23 group treated with hemin and NF-κB inhibitor. D Relative intensity of TLR4, MyD88 and P65 in BV2 cells after treatment for 24 h. ELISA analysis of TNF-α E, IL-1β F and IL-6 G in BV2 cells after treatment for 24 h. H Representative immunofluorescence images of CD86 (red) staining in BV2 cells. Scale bar = 100 μm. I Quantitative analysis of CD86 + cells. Data are presented as means ± SEM. ***p < 0.001. **p < 0.01. *p < 0.05. ns p > 0.05