Fig. 7

Disruption of the C/EBPβ-Clec7a axis attenuated neuropathic pain by regulating NLRP3 inflammasome-induced pyroptosis in vivo. A Timeline schematic of experimental paradigm. B The decreased expression of C/EBPβ significantly ameliorated mechanical withdrawal threshold in CCI rat model, while overexpression Clec7a facilitated neuropathic pain (Sham group, n = 9; the other groups, n = 15 rats/group). C Expression levels of C/EBPβ, Clec7a, pSyk, pERK, pJNK, NLRP3 and GSDMD proteins in ipsilateral spinal cord (SC) of CCI rats on postoperative 4 weeks were measured using western blot analysis (n = 3 independent blots). D Immunohistochemical detection of C/EBPβ and Clec7a in ipsilateral SC of CCI rats after treatment (n = 9 rats/group, 1 section/rat). White arrows point to C/EBPβ- or Clec7a-positive cells, respectively. Scale bar: 100 μm (20 ×); 50 μm (40 ×). E The concentration of IL-1β and IL-18 in serum of indicated groups were assessed by ELISA (n = 3 rats/group with 3 technical replicates). F Representative images (up) and quantification evaluation (down) of Tunel staining in the indicated groups (n = 6 rats/group, 3 section/rat). Red: Tunel-positive cells; blue: nuclei (Hoechst 33,258). Scale bar: 100 μm. Data are presented as mean ± SEM. Two-way repeated measures ANOVA and Tukey’s post hoc test was used to analyze data at different time points B. One-way ANOVA was used to analyze data among multiple groups followed by Tukey’s post hoc test for equal variances or Dunnett T3 post hoc test for unequal variances C–F. ^**P < 0.01, ^***P < 0.001 compared with the Sham group; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 compared with CCI + pLV.1-NC + pLVSO5-NC group; ^△P < 0.05, ^△△P < 0.01, ^△△△P < 0.001 compared with CCI + pLV.1-shCebpb + pLVSO5-NC group; ^§P < 0.05, ^§§P < 0.01, ^§§§P < 0.001 compared with CCI + pLV.1-NC + pLVSO5- Clec7a-overexpress group