Figure 8.

AAV9-HMGB1 attenuates the blocking effect of M2φ-Evs on the TLR4-NF-κB signaling pathway. Adenovirus overexpression vector harboring HMGB1 (AAV9-HMGB1) or empty vector (AAV9-NC) was injected into M2φ-Evs-treated mice via intrathecal injection. (A) The expression of HMGB1 and TLR4 and the phosphorylation levels of IKKB as well as NF-κB p65 in spinal cord tissues examined by western blot; (B) Mean (± SD) PWMT (in g) in response to five von Frey hair stimulations; (C) NSF in a time span of 2 min; (D) ELISA detection of inflammatory factors TNF-α, IL-1β, IL-6 in mouse serum; (E, F) Expression of the glial activation marker GFAP in mouse spinal cord tissues was detected by RT-qPCR (E) and immunofluorescence (F). n = 8. **P < 0.01 as determined by unpaired t test (D-F) or two-way ANOVA (A-C), followed by Tukey’s post-hoc analysis.