Fig. 2
From: miRNA-23a/CXCR4 regulates neuropathic pain via directly targeting TXNIP/NLRP3 inflammasome axis
miR-23a regulates CXCR4 expression in spinal astrocyte in neuropathic pain. a The shared miRNA numbers to targeting CXCR4 predicted by the use of TargetScan and MicroRNA program. b The informatics analysis of miR-23a binding the 3′UTR in CXCR4 mRNA. c Time course of spinal miR-23a expression in pSNL-induced chronic neuropathic pain mice. One-way ANOVA (expression versus time point) followed by post hoc Tukey test, Ftime (5, 24) = 19.66, *p < 0.05, **p < 0.01 versus sham group. d In vitro validation of miR-23a targeting CXCR4. A mutation was generated in the CXCR4-3′-UTR mRNA sequence in the complementary site for the seed region of miR-23a as indicated (CHK-mut-CXCR4). **p < 0.01 versus the corresponding CHK-mut-CXCR4 or empty vector group by two-tailed unpaired Student’s t test. e The validation of transfection efficiency of miR-23 mimics or Lenti-miR-23a in the mouse spinal cord by qRT-PCR. Spinal cord was harvested 24 h after intrathecal injection of continuous 2-day miR-23a mimics in naïve mice or pSNL mice with 7-day surgery or 72 h after intrathecal injection of continuous 2-day Lenti-miR-23a in naïve mice or pSNL mice with 7-day surgery. f, g The increased spinal CXCR4 protein expression in pSNL mice was reversed by intrathecal injection of miR-23a mimics (f) or Lenti-miR-23a (g). Intrathecal injections of miR-23a mimics or Lenti-miR-23a were performed from day 7 after pSNL. CXCR4 was measured at 24 h after 2-day miR-23a mimics injections or 48 h after 3-day Lenti-miR-23a injections. One-way ANOVA (expression versus treatment groups) followed by post hoc Tukey test, f F (3, 16) = 11.2, g Ftime (3, 16) = 14.6, **p < 0.01, ***p < 0.001, sham group; #p < 0.05 versus pSNL+Scr or pSNL+Lenti-vector group. h, i Spinal CXCR4 protein expression was increased by intrathecal injection of miR-23a inhibitor (miR-23a Ih) (h) or LV-miR-23a (LV-23a) (i) in naïve mice. CXCR4 was measured at 24 h after 2-day miR-23a inhibitor injections or at 48 h after 3-day LV-miR-23a injections. One-way ANOVA (expression versus treatment groups) followed by post hoc Tukey test, h G (2, 12) = 26.98, i H (2, 12) = 24.56, *p < 0.05, **p < 0.01 versus Scr or vector group. Data are presented as mean ± SEM; n = 5 per group