Sepiapterin Improves Vascular Reactivity and Insulin‐Stimulated Glucose in Wistar Rats

Figures

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  Figure 1. Insulin and glucose intraperitoneal tolerance test. Animal insulin and glucose tolerance were assessed at baseline and end of the study (sac). Control group n = 5, SEP n = 4 at baseline, control group n = 5, and SEP n = 3 at endpoint. ∗Interaction effect of p < 0.05; ∗∗difference at sacrifice as determined by post hoc analysis (p < 0.05) and repeated-measures two-way ANOVA.

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  Figure 2. (a, b, and c) Aorta vasodilation. At the end of the study, aorta rings (2 mm) were hung in an upright water muscle bath and analyzed for vasodilation using a dose response to ACh (a) and NO donor S-nitroso-N-acetyl-DL-penicillamine (SNAP, b) and cGMP (c) in intact and denuded endothelium. Control group n = 5 and SEP n = 4. Data are presented as mean ± SEM, ∗p < 0.05, Student's t-test.

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  Figure 3. (a and b) Aorta vasoconstriction. At the end of the study, aorta rings (2 mm) were hung in an upright water muscle bath and analyzed for vasoconstriction using high potassium buffer (a) and a dose response to phenylephrine (PE, b). Control group n = 5 and SEP n = 4. Data are presented as mean ± SEM, Student's t-test.

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  Figure 4. (a–d). Mitochondrial respiration. Aorta from animals were permeabilized and exposed to two different experiments using various substrates (a, b: pyruvate, malate, glutamate, ADP, cytochrome c, succinate, oligomycin, and carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone (FCCP)) or (c, d: octanoylcarnitine, malate, glutamate, ADP, cytochrome c, succinate (S), oligomycin, and (FCCP)). State 2 respiration (S2), state 3 respiration (S3), and state 4 respiration (S4) were measured. States were analyzed as shown above, including respiratory control ratio (RCR, state 3(S)/state 4). Control group n = 5 and SEP n = 4. Data are presented as mean ± SEM, ∗p < 0.05, Student's t-test.

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  Figure 5. (a–d) Cellular signaling in aorta tissue: aorta tissue was harvested and lysates processed for Western blot analysis. Blots of aorta were probed for pAMPK, AMPK (a), peNOS, eNOS (b), PGC-1α, SIRT3, MnSOD (c), and mitochondrial complexes (d). In (d), complex I was calculated using enhanced blot image for visibility and was found not to be significantly different from control (data not shown on blot image). Ladder (Ld) was loaded to orient protein identification, and β-actin was used for normalization. Specific activity was calculated as phosphorylated protein expression normalized to total protein expression. Control group n = 5 and SEP n = 4. Data are presented as mean ± SEM, ∗p < 0.05, Student's t-test.