Fig. 7: Decreased β-EP mediates mechanical allodynia in CCI-ION rats. | Nature Communications

Fig. 7: Decreased β-EP mediates mechanical allodynia in CCI-ION rats.

From: Epigenetic regulation of beta-endorphin synthesis in hypothalamic arcuate nucleus neurons modulates neuropathic pain in a rodent pain model

Fig. 7

a Schematic diagram showing that PC1 cleaves POMC to β-LPH, and PC2 further cleaves β-LPH to β-EP. b, c Representative immunoblots (b) and bar charts (c) indicate the protein abundance of POMC in the ARC. **p < 0.01 versus POMC on Day 0, one-way ANOVA followed by Bonferroni’s test. n = 8 rats per time point per group. d, e ELISA analysis of β-LPH (d, n = 4 rats per group, p = 0.0001) or β-EP (e, n = 6 rats per group, p = 0.0002). ***p < 0.001 versus sham, two-sided Student’s t-test. f Immunostaining analysis of PC1 (red) with β-EP (green) and DAPI (blue) in the ARC. Bar charts show the percentage of both PC1+ and β-EP + ARC neurons among total numbers of DAPI-labeled cells. Arrows show colocalization. **p = 0.0031 versus sham, two-sided Student’s t-test. n = 4 rats per group. Scale bar, 25 μm. g–i Administration of PCSK1-up (g), HDAC9-up (h), or antagomir-203a (i) abolished the CCI-ION-induced β-EP decrease in the ARC. **p < 0.01, ***p < 0.001 versus sham, ##p = 0.0027 versus CCI-ION + PCSK1-NC, ++p = 0.0023 versus CCI-ION + HDAC9-NC, @@p = 0.0021 versus CCI-ION + antagomir-NC, one-way ANOVA followed by Bonferroni’s test. n = 6 rats per group. j–l Administration of PCSK1-siRNA (j), HDAC9-siRNA (k), or agomir-203a (l) decreased the content of β-EP in the ARC of intact rats. **p = 0.0037, ***p = 0.0002 versus NC-siRNA, ###p = 0.0005 versus agomir-NC, one-way ANOVA followed by Bonferroni’s test. n = 6 rats per group. m, Evaluation of POMC-siRNA or NC-siRNA on the miR-203a-down-mediated attenuation of mechanical allodynia in CCI-ION rats. ***p < 0.001 versus sham + NC-siRNA, ###p < 0.001 versus CCI-ION + POMC-siRNA, two-way ANOVA followed by Bonferroni’s test. n = 8 rats per group. n Intravenous administration of CTAP (0.1 mg/ml) completely abolished the miR-203a-down-induced alleviation of mechanical allodynia in CCI-ION rats. ***p < 0.001 versus miR-NC + vehicle at Day 0, ###p < 0.001 versus miR-203a-down + vehicle, two-way ANOVA followed by Bonferroni’s test. n = 8 rats per group. All data are presented as mean values ± SEM.

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