Fig. 6: PC1 is responsible for miR-203a-3p-mediated nociceptive behaviors.
a, b Time course of PC1 and PC2 protein expression in the rat ARC after CCI-ION (a) or sham surgery (b). ***p < 0.001 versus PC1 on Day 0, one-way ANOVA followed by Bonferroni’s test. n = 8 rats per time point per group. c Correlation between miRNA and PCSK1 mRNA expression (R2 = 0.8113). d Immunostaining analysis of PC1 (red) with NeuN, GFAP, and CD11b (green) in ARC sections of intact rats. Bar charts show the percentage of double-stained ARC neurons among total numbers of PC1+-labeled neurons. Arrows show colocalization. n = 4 rats per group. Scale bar, 25 μm. e Administration of lenti-hSyn-PCSK1-up (PCSK1-up) restored the CCI-ION-induced decrease in PC1 expression. ***p = 0.0001 versus CCI-ION + PCSK1-NC, two-sided Student’s t test. n = 5 rats per group. f, g Injection of PCSK1-up alleviated CCI-ION-induced mechanical allodynia in male (f) or female (g) rats. ***p < 0.001 versus sham + PCSK1-NC, ###p < 0.001 versus CCI-ION + PCSK1-NC, two-way ANOVA followed by Bonferroni’s test. n = 8 rats per group. h PCSK1-up pretreatment abolished the CCI-ION-induced decrease in mechanical threshold. ***p < 0.001 versus PCSK1-NC + sham, ###p < 0.001 versus PCSK1-NC + CCI-ION, two-way ANOVA followed by Bonferroni’s test. n = 8 rats per time point per group. i Bilateral administration of PCSK1-up alleviated SNL-induced mechanical allodynia. ***p < 0.001 versus sham + PCSK1-NC, ###p < 0.001 versus SNL + PCSK1-NC, two-way ANOVA followed by Bonferroni’s test. n = 8 rats per group. j Administration of PCSK1 siRNA (PCSK1-siRNA) reduced PC1 expression in the ARC of intact rats. ***p = 0.0003 versus NC-siRNA, one-way ANOVA followed by Bonferroni’s test. n = 4 rats per group. k Application of PCSK1-siRNA in intact rats induced mechanical hypersensitivity. **p < 0.01, ***p < 0.001 versus NC-siRNA, two-way ANOVA followed by Bonferroni’s test. n = 10 rats per group. l, m Effects of miR-203a-down (l) or HDAC9-up (m) on the decreased PC1 expression induced by CCI-ION. ***p = 0.0006 versus CCI-ION + miR-NC, ###p = 0.0006 versus CCI-ION + HDAC9-NC, one-way ANOVA followed by Bonferroni’s test. n = 4 rats per group. n Evaluation of preinjection of PCSK1-up on the miR-203a-down-induced attenuation of mechanical allodynia in CCI-ION rats. ***p < 0.01 versus sham + PCSK1-NC, ###p < 0.001 versus CCI-ION + PCSK1 on Day 7, +++p < 0.001 versus CCI-ION + PCSK1-up on Day 0, two-way ANOVA followed by Bonferroni’s test. n = 9 rats per group. All data are presented as mean values ± SEM.
