Fig. 4
From: miRNA-23a/CXCR4 regulates neuropathic pain via directly targeting TXNIP/NLRP3 inflammasome axis
TXNIP is involved in the process of neuropathic pain by the CXCR4-dependent regulation. a Quantitative expression of Txnip mRNA at 1, 3, 7, 14, and 21 days after pSNL surgery. One-way ANOVA (expression versus time point) followed by post hoc Tukey test, Ftime (5, 24) = 40.4, *p < 0.05, **p < 0.01 versus sham group; n = 5 per group. b–d TXNIP immunofluorescent co-staining with NeuN (b and b′), GFAP (c and c′), or IBA1 (d and d′) in the lumbar of spinal cord at 7 days after pSNL surgery. *p < 0.05, **p < 0.01 versus sham group by two-tailed unpaired Student’s t test; n = 5 per group. Scale bar, 50 μm. e Daily intrathecal injections of TXNIP siRNA for 3 consecutive days reversed pSNL-induced thermal hyperalgesia and mechanical allodynia. Two-way ANOVA (effect versus group × time interaction) followed by post hoc Tukey test, PWL Fgroup (10, 72) = 20.76; PWT Fgroup (10, 72) = 6.85, *p < 0.05, **p < 0.001, versus pSNL+Scr group; n = 5 per group. f, f′ Co-staining of double immunofluorescence (CXCR4, green; TXNIP, red) in the lumbar segment of the spinal cord of naïve mice. Scale bar, 50 μm. g The representative immunoblot demonstrates the interaction of TXNIP and CXCR4 in a co-IP experiment. Rabbit IgG is used as control for co-IP assays. **p < 0.01, ***p < 0.001 versus the corresponding sham group by two-tailed unpaired Student’s t test; n = 5 per group. h The CDS region of gene Txnip was inserted into pBIND to produce Gal4-Txnip chimeric fusion expression vector (pBIND-Txnip), and the C-terminal of Cxcr4 was inserted into pACT to produce VP16-Cxcr4-C chimeric fusion expression vector (pACT-CXCR-C). i pBIND-Txnip and/or pACT-Cxcr4-C transfected into 293T cells with the reporter gene vector pG5luc or empty vector. At 48 h after transfection, the interactions were measured by relative luciferase activity. In general, only pBIND-Txnip and pACT-Cxcr4-C co-transfection with pG5luc indicated strong luciferase activity. One-way ANOVA (expression versus the treated groups) followed by post hoc Tukey test, F (5, 24) = 628, ***p < 0.001; n = 3 per group