Fig. 6

TXNIP regulates the protein expression of spinal NLRP3 inflammasome in SNL-induced neuropathic pain. a Co-IP of TXNIP and NLRP3. Rabbit IgG is used as control for co-IP assays. *p < 0.05, **p < 0.01 versus the corresponding sham group by two-tailed unpaired Student’s t test; n = 5 per group. b Increased NLRP3 inflammasome complex after pSNL, including NLRP3, ASC, P-Caspase1, C-Caspase1, and mature IL-1β, were reversed by knockdown of TXNIP with siRNA. Inflammasome complex expression was determined on day 7 after pSNL or at 24 h after 3-day injections of TXNIP siRNA or Scr, beginning on day 7 after pSNL. One-way ANOVA (expression versus the treated groups) followed by post hoc Tukey test, F (3, 8) = 76.56 for TXNIP, 106.9 for NLRP3, 79.01 for ASC, 191.3 for P-Caspase1, 26.68 for C-Caspase1, and 33.34 for mature IL-1β, **p < 0.01, ***p < 0.001 versus sham group. ^#p < 0.05, ^##p < 0.01, and ^###p < 0.001 versus pSNL+Scr group. c Increased NLRP3 inflammasome were reversed by intrathecal injection of Lenti-miR-23a in pSNL mice. Inflammasome complex expressions were measured at 7 days after pSNL surgery or at 48 h after 3-day injections of Lenti-miR-23a or Lenti-vector, beginning on day 7 after pSNL. One-way ANOVA (expression versus the treated groups) followed by post hoc Tukey test, F (3, 8) = 20.79 for NLRP3, 96.78 for ASC, 29.35 for P-Caspase1, 106.1 for C-Caspase1, and 42.21 for mature IL-1β, **p < 0.01, ***p < 0.001 versus sham group. ^#p < 0.05, ^##p < 0.01, and ^###p < 0.001 versus pSNL+Lenti-miR-23a group. d LV-miR-23a-induced expression of NLRP3 inflammasome was reversed by knockdown of TXNIP with siRNA in naïve mice. Content of inflammasome was examined at 48 h after 3-day injections of LV-miR-23a or at 24 h after 3-day injections of TXNIP siRNA or Scr (starting after the injections LV-miR-23a or vector). One-way ANOVA (expression versus the treated groups) followed by post hoc Tukey test, F (3, 8) = 106.7 for NLRP3, 138.4 for ASC, 54.04 for P-Caspase1, 133 for C-Caspase1, and 50.59 for mature IL-1β, **p < 0.01, ***p < 0.001 versus vector group. ^#p < 0.05, ^##p < 0.01, and ^###p < 0.001 versus LV-miR-23a + Scr group. e Increased NLRP3 inflammasome induced by injection of Lenti-CXCR4 was reversed by injection of TXNIP siRNA in naïve mice. Inflammasome complex expressions were measured at 48 h after 3-day injections of Lenti-CXCR4 or at 24 h after 3-day injection of TXNIP siRNA or Scr (starting after injections of Lenti-CXCR4 or Lenti-vector). One-way ANOVA (expression versus the treated groups) followed by post hoc Tukey test, F (3, 8) = 18.84 for NLRP3, 48.48 for ASC, 84.25 for P-Caspase1, 8.63 for C-Caspase1, and 48.37 for mature IL-1β, **p < 0.01, ***p < 0.001 versus Lenti-vector group. ^#p < 0.05, ^##p < 0.01, and ^###p < 0.001 versus Lenti-CXCR4 + Scr group. f NLRP3 inflammasome upregulated by injection of LV-miR-23a was reversed by injection of CXCR4 siRNA in naïve mice. One-way ANOVA (expression versus the treated groups) followed by post hoc Tukey test, F (3, 8) = 42.41 for NLRP3, 72.1 for ASC, 10.9 for P-Caspase1, 82.57 for C-Caspase1, and 290.7 for mature IL-1β, **p < 0.01, ***p < 0.001 versus vector group. ^#p < 0.05, ^##p < 0.01 versus LV-miR-23a + Scr group. Expression of inflammasome was measured at 48 h after 3-day injections of LV-miR-23a or at 6 h after injection of CXCR4 siRNA or Scr (beginning after injections of LV-miR-23a or Vector). Data are presented as mean ± SEM; n = 3 per group