Fig. 5

Inhibition of Sirt3/Pgc-1α partially blocked the effect of SS-31 on headache treatment. A Schematic diagram of the experiment. C57BL/6 mice received PBS + sham + DMSO, PBS + IS + DMSO, SS-31 + IS + DMSO, SS-31 + IS + 3-TYP or SS-31 + IS + SR-18292 treatment for 7 consecutive days, followed by behavioral tests and sacrifice to evaluate nociceptive responses. B-D The number of head scratching in 1 h (F(4, 40) = 9.107, ^****p < 0.0001; ^****p_{(PBS+IS+DMSO)} < 0.0001, ⁺⁺⁺p_{(SS-31+IS+DMSO)} = 0.0001, ⁺p_{(SS-31+IS+3-TYP)} = 0.015, ⁺p_{(SS-31+IS+SR-18292)} = 0.0475, periorbital mechanical threshold (F(4, 40) = 9.986, ^****p < 0.0001; ^***p_{(PBS+IS+DMSO)} = 0.0001, ^***p_{(SS-31+IS+3-TYP)} < 0.0004, ^*p_{(SS-31+IS+SR-18292)} = 0.0260, ⁺⁺p_{(SS-31+IS+DMSO)} = 0.0011, ^##p_{(SS-31+IS+3-TYP)} = 0.0031) and paw withdrawal latency (F(4, 40) = 8.679, ^****p < 0.0001; ^***p_{(PBS+IS+DMSO)} = 0.0003, ^**p_{(SS-31+IS+3-TYP)} = 0.0027, ^**p_{(SS-31+IS+SR-18292)} = 0.0015, ⁺⁺p_{(SS-31+IS+DMSO)} = 0.0080, ^#p_{(SS-31+IS+3-TYP)} = 0.0464, ^#p_{(SS-31+IS+SR-18292)} = 0.0288) were recorded in different groups. n = 9 per group; One-way ANOVA. E–G Western blot analysis (n = 9 per group; F(4, 40) = 6.389, ^***p = 0.0004; ^**p_{(PBS+IS+DMSO)} = 0.0058, ^*p_{(SS-31+IS+SR-18292)} = 0.0451, ⁺⁺p_{(SS-31+IS+DMSO)} = 0.0022, ^#p_{(SS-31+IS+SR-18292)} = 0.0195) and immunofluorescence staining (n = 6 per group; F(4, 25) = 10.36, ^****p < 0.0001; ^***p_{(PBS+IS+DMSO)} = 0.0005, ^**p_{(SS-31+IS+3-TYP)} = 0.0017, ^*p_{(SS-31+IS+SR-18292)} = 0.0286, ⁺⁺⁺p_{(SS-31+IS+DMSO)} = 0.0009, ^##p_{(SS-31+IS+3-TYP)} = 0.0029, ^#p_{(SS-31+IS+SR-18292)} = 0.0446) illustrated that expression levels of c-fos in TNC of different groups. Scale bar, 50 μm. One-way ANOVA. H Immunofluorescence staining (F(4, 25) = 14.31, ^****p < 0.0001; ^****p_{(PBS+IS+DMSO)} < 0.0001, ^***p_{(SS-31+IS+3-TYP)} = 0.0003, ^****p_{(SS-31+IS+SR-18292)} = 0.0004, ⁺⁺⁺p_{(SS-31+IS+DMSO)} = 0.0009, ^#p_{(SS-31+IS+SR-18292)} = 0.0425) was used to examine the levels of CGRP in TNC of different groups. Scale bar, 200 μm. n = 6 per group; One-way ANOVA. Data are represented as Mean ± SD; ^*p < 0.05, ^**p < 0.01 and ^***p < 0.001 as compared to PBS + Sham + DMSO group. ⁺p < 0.05, ⁺⁺p < 0.01 and ⁺⁺⁺p < 0.001 as compared to PBS + IS + DMSO group. ^#p < 0.05 as compared to SS-31 + IS + DMSO group