Effects of different diets used in diet-induced obesity models on insulin resistance and vascular dysfunction in C57BL/6 mice

Figures

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  Figure 1. Development of the bodyweight. Male C57BL/6J mice were put on standard chow diet (SCD), cafeteria diet (CAF), normal-fat diet (NFD) or high-fat diet (HFD) at the age of 6 weeks for another 12 weeks. Food intake per day per mouse in gram (A) and changes in food consumption per week per mouse in gram over the feeding period (B) are pictured. ANOVA followed by Fisher’s protected least significant difference test as post hoc method was performed to compare all four groups. Panel C shows the development of the bodyweight over time. Panel D shows the bodyweight after 12 weeks of experimental feeding. Significances marked with *,# and +refer to comparisons to the SCD, CAF and NFD group, respectively. *,#P < 0.05; ***,###,+++P < 0.001; ns, not significant. n = 15 (SCD and CAF) or 20 (NFD and HFD), respectively.

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  Figure 2. Weight of fat mass. Male C57BL/6J mice were put on experimental diets at the age of 6 weeks for another 12 weeks. The weights of epididymal (A), lumbar (B) and mesenteric fat pads (C) at the end of the experiment are shown. Significances marked with *, # and +refer to comparisons to the SCD, CAF and NFD group, respectively *P < 0.05; **P < 0.01; ***###,+++P < 0.001; ns, not significant. n = 15 (SCD and CAF) or 20 (NFD and HFD), respectively.

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  Figure 3. Glucose intolerance. Male C57BL/6J mice were put on experimental diets at the age of 6 weeks for another 12 weeks. Panel A shows the fasting glucose levels in the 12th week of feeding. n = 15 (SCD and CAF) or 20 (NFD and HFD), respectively. Panel B shows the plasma insulin concentration in the 12th week of feeding. n = 6 (SCD, NFD and HFD) or 5 (CAF), respectively. Panel C shows the results of GTT at the end of the experiment after overnight fasting. n = 5 (SCD, CAF and HFD) and 9 (NFD), respectively. Panels E shows the results of ITT (1 IU/kg body weight insulin was injected intraperitoneally) in the 12th week of experimental feeding after overnight fasting. n = 10 (SCD and NFD) and 9 (CAF and HFD), respectively. The area under the curve (AUC) of GTT and ITT are shown in panels D and F, respectively. Significances marked with *,# and +refer to comparisons to the SCD, CAF and NFD group, respectively. *,#,+P < 0.05, **,##P < 0.01, ***,###,+++P < 0.001, ns, not significant.

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  Figure 4. Vascular function. Male C57BL/6J mice were put on experimental diets at the age of 6 weeks for another 12 weeks. Vasodilation was induced by acetylcholine in norepinephrine-pre-contracted aorta in myograph experiments, without PVAT (A), with PVAT (B–E), or in the presence of NO synthase inhibitor L-NAME (E). *P < 0.05, **P < 0.01, ***P < 0.001. n = 9 (SCD and CAF) and 15 (NFD and HFD), respectively.

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  Figure 5. Phosphorylation of eNOS. Male C57BL/6J mice were put on experimental diets at the age of 6 weeks for another 12 weeks. Phosphorylation of eNOS at Ser1177 (A) and of Akt at Ser473 (B) in aortic PVAT was studied by Western blot analyses. The blots shown are representative of four independent experiments with similar results. Significances marked with *,# and +refer to comparisons to the SCD, CAF and NFD group, respectively. *,#P < 0.05, **,##,++P < 0.01, ***P < 0.001, ns, not significant. n = 12.

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  Figure 6. Coagulation. Male C57BL/6J mice were put on experimental diets at the age of 6 weeks for another 12 weeks. Prothrombin time (PT) (A) and activated partial thromboplastin time (APTT) (B) were measured with citrate plasma on a KC4 Delta Amelung Coagulometer. Significances marked with *,# and +refer to comparisons to the SCD, CAF and NFD group, respectively. *P < 0.05, ***P < 0.001, ns, not significant. n = 10 (SCD and CAF), 9 (NFD) and 11 (HFD), respectively.

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  Figure 7. Activity of coagulation factors. Male C57BL/6J mice were put on experimental diets at the age of 6 weeks for another 12 weeks. The activity of coagulation factors was determined using deficient plasma (shorter coagulation time means higher activity) on a KC4 Delta Amelung Coagulometer. Shorter coagulation time indicates higher activity. Significances marked with *,# and +refer to comparisons to the SCD, CAF and NFD group, respectively. *,#,+P < 0.05, **P < 0.01, ***P < 0.001, ns, not significant. n = 9 (SCD, NFD and CAF) and 11 (HFD), respectively.