Fig 1: CB1 receptors in the striate muscle and AM251 effects on the mitochondrial CB1 receptor and DLD.Electron immunogold detection of CB1 receptors (arrows) on the mitochondrial (mt) membrane of striate muscle cells in rats (A,B), and wild-type (C,D) and CB 1 -/- (E,F) mice. Scale bars: 0.5μm. AM251 effects on the protein expression of CB1 receptor (G) and DLD (H) in the purified mitochondrial fraction of the abdominal muscle of STD, HFD and HCD-fed rats. Representative immunoblots (I). Bonferroni’s test (n = 8): *P<0.05, **P<0.01 vs. STD-vehicle; $ P<0.05 vs. HCD-vehicle.
Fig 2: Scheme showing the enzymatic reactions that catalyze the pyruvate dehydrogenase complex, including pyruvate dehydrogenase activity and diaphorase activity of DLD (A) as well as the glutathione reductase/peroxidase activity (B).NBT staining is proportional to O- production from redox activity.
Fig 3: AM251 regulates the expression of DLD and Cox4i1.AM251 effects on the protein and gene expressions of DLD (A,B) and the gene expression of Cox4i1 (C) in the abdominal muscle of STD, HFD and HCD-fed rats. Representative 2D polyacrylamide gels showing the intensity of the identified spots for DLD (D). Bonferroni’s test (n = 6–8): *P<0.05, **P<0.01, ***P<0.001 vs. STD-vehicle; # P<0.05 vs. HFD-vehicle; $ P<0.05 vs. HCD-vehicle. Gene (D) and protein (E) expressions of DLD in the abdominal muscle of wild-type and CB 1 -/- mice. Representative immunoblots (F). Student’s t test (n = 6): *P<0.05, ***P<0.001 vs. WT.
Fig 4: AM251 increased diaphorase/oxidative, pyruvate dehydrogenase and glutathione reductase activity.AM251 and ACEA effects on the diaphorase/oxidative activity in C2C12 myotubes in culture (A,B), and in the C2C12 mitochondrial fraction in vitro (I). AM251 and Representative images of differentiated C2C12 cells showing NBT staining (C-G). Representative immunoblots of CB1 receptor and DLD from the total and mitochondrial protein fractions of C2C12 myotubes (H). ACEA effects on the pyruvate dehydrogenase, glutathione content and glutathione reductase activity in C2C12 myotubes in culture (J-L). ANOVA (n = 8): *P<0.05, **P<0.01, ***P<0.001 vs. control C2C12 (white bar); & P<0.05, &&& P<0.001 vs. insulin C2C12 (black bar).
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