Figure 8

Addition of recombinant High-mobility group box 1 (rHMGB1) in the subarachnoid space triggered inflammatory response in vivo . NF-κB was measured by the western blot of its P65 subunit in the nucleus. Toll-like receptor (TLR)4 protein level was also detected by western blot analysis. IL-1β was measured by real-time PCR. (A) rHMGB1 up-regulated P65 subunit protein level in the nuclear protein in cortex cells; P < 0.05 between the 12-h, day-1 group and control group, P < 0.01 between the day-2 group and control group. (B) rHMGB1 increased TLR4 protein level. P < 0.05 between the 12-h and control group, P < 0.01 between the day-1, day-2 group and control group. (C) rHMGB1 upregulated IL-1β mRNA expression in cortex cells: P < 0.05 between the 12-h, day-2 group and control group, P < 0.01 between the day-1 group and control group. (D) Western blot analysis of histone 3 content in rHMGB1(left lane) and the nuclear protein extraction(right lane). The result could exclude histone 3 contamination in rHMGB1 products. Histone 3 was predominant composition of histone protein, thus, our result indicated the rHMGB1 used in the study had good purity. Bars represent the mean ± standard error (n = 6): *P < 0.05 compared with the control group; ^# P < 0.01 compared with the control group.