Fig. 3

SS-31 restored mitochondrial function and mitochondrial homeostasis in an IS-induced headache mouse model. C57BL/6 mice received sham, IS or SS-31 treatment for 7 consecutive days, and were sacrificed to evaluate the effects of SS-31 on mitochondrial function and mitochondrial homeostasis in TNC. A-C Western blot analysis was used to asses expression levels of Pgc-1α (F(2, 15) = 5.434, ^*p = 0.0168; ^*p = 0.0384, ⁺p = 0.0249), Tfam (F(2, 15) = 6.457, ^**p = 0.0095; ^*p = 0.0443, ⁺⁺p = 0.0100), Mfn2, Drp1, Fis1 (F(2, 15) = 15.74, ^***p = 0.0002; ^**p = 0.0015, ⁺⁺⁺p = 0.0003), P62 and Pink1 (F(2, 15) = 5.143, ^*p = 0.0199; ^*p = 0.0153) in different groups. n = 6 per group; One-way ANOVA. D Western blot analysis and quantification of Sirt3 (F(2, 15) = 10.66, ^**p = 0.0013; ^**p = 0.0011, ⁺p = 0.0208) protein level normalized to β-actin. n = 6 per group; One-way ANOVA. E The levels of ROS (F(2, 15) = 25.33, ^****p < 0.0001; ^****p < 0.0001, ⁺⁺⁺p = 0.0003), ATP (F(2, 15) = 7.704, ^**p = 0.0050; ^*p = 0.0413, ⁺⁺p = 0.0045), MMP (F(2, 15) = 25.89, ^****p < 0.0001; ^****p < 0.0001, ⁺⁺⁺p = 0.0009) and MDA (F(2, 15) = 3.854, ^*p = 0.0446; ⁺p = 0.0362) were detected and normalized by total protein concentrations in different groups. n = 6 per group; One-way ANOVA. (F) Mitochondrial ultrastructure in TNC by TEM analysis. The abnormal mitochondria count percent (F(2, 15) = 14.39, ^***p = 0.0003; ^***p = 0.0004, ⁺⁺p = 0.0034) and area percent (F(2, 15) = 13.70, ^***p = 0.0004; ^***p = 0.0003, ⁺p = 0.0120) were calculated. Red arrowhead, damaged mitochondria (swollen mitochondria and reduction of mitochondrial cristae). Black arrowhead, normal mitochondria. Scale bar, 500 nm. n = 6 per group; One-way ANOVA. Data are represented as Mean ± SD; ^*p < 0.05, ^**p < 0.01, ^***p < 0.001 and ^****p < 0.0001 as compared to sham group. ⁺p < 0.05, ⁺⁺p < 0.01 and ⁺⁺⁺p < 0.001 as compared to IS group