Fig. 6: RFGF21 activates CaMKK2 to upregulate AMPKα activity, improve endothelium-dependent relaxation and alleviate oxidative stress in aortas challenged with HG.
a Aortas isolated form C57BL/6J mice were pretreated with FIIN-4 (10 μM) for 30 mins and exposed to either HG (30 mM) alone or HG plus rFGF21 (0.01 mg/ml) for an additional 2 h. Phosphorylation level of CaMKK2 as determined by western blot analysis (upper panel) and quantitation using ImageJ software (lower panel) (n = 4). b–g Aortas isolated form C57BL/6J mice were pretreated with STO-609 (5 μg/ml) for 30 mins and exposed to either HG (30 mM) alone or HG plus rFGF21 (0.01 mg/ml) for an additional 2 h. b Dose-dependent relaxation to ACh (n = 5). c NO2 and NO3 levels stimulated by ACh (6 × 10−8 M) for 3 mins (n = 5). d Phosphorylation level of eNOS stimulated by ACh (6 × 10−8 M) for 3 mins as determined by western blot analysis (upper panel) and quantitation using ImageJ software (lower panel) (n = 4). e Immunofluorescent DHE staining (n = 6). The upper panel shows DHE staining and the lower panel shows quantitation using ImageJ software. Scale bars, 100 μm. (f, g) Phosphorylation level of AMPKα (f) and ACC (g) as determined by western blot analysis (upper panel) and quantitation using ImageJ software (lower panel) (n = 4). All data are presented as mean ± SEM. *p < 0.05 vs Control; #p < 0.05 vs HG; †p < 0.05 vs HG + FGF21
