Impact of Prenatal Exposure to Maternal Diabetes and High-Fat Diet on Postnatal Myocardial Ketone Body Metabolism in Rats

Figures

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  Figure 1. Respiratory capacity increased with increasing concentration of βHOB in both control and combination exposed cardiomyocytes. (A) Basal respiration, (B) Ketone oxidation, (C) Maximal respiration, and (D) Spare respiratory capacity in control and combination exposed NRCM. Bars represent mean OCR (n = 10–11 litter per group). Blue color indicates controls and red color indicates combination exposed. Open symbols in the graph indicate males and filled symbols indicate females. Ketone effect is noted with p values, as shown in the figures; there was no group or interaction effect, as determined by two-way ANOVA. mM refers to the progressive concentrations of βHOB supplied.

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  Figure 2. Schematic representation of the ketone stress test (KST) for assessing glycolysis in terms of extracellular acidification rate (ECAR).

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  Figure 3. Ketones improve glucose mediated and anaerobic ECAR in combination exposed, but not in control, NRCM. (A) Ketone mediated ECAR, (B) Maximal ECAR, (C) Glucose mediated ECAR, and (D) Anaerobic ECAR in control and combination exposed NRCM. Bars represent mean ECAR (n = 10–11 litter per group). Blue color indicates controls and red color indicates combination exposed. Open symbols in the graph indicate males and filled symbols indicate females. Interaction effect was determined by two-way ANOVA, and the group differences remain significant only in combination exposed NRCM by one-way ANOVA with p values, as shown in the figures. mM refers to the progressive concentration of βHOB supplied.

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  Figure 4. Maximal and glucose mediated total proton efflux rate (PER) tended to increase with increasing concentrations of βHOB. (A) Ketone mediated PERtotal, (B) Maximal PERtotal, (C) Glucose mediated PERtotal, and (D) Anaerobic PERtotal in control and combination exposed NRCM. Bars represent mean PER (n = 10–11 litter per group). Blue color indicates controls and red color indicates combination exposed. Open symbols in the graph indicate males and filled symbols indicate females. Ketone mediated effect was determined by two-way ANOVA with p values as shown. mM refers to the progressive concentration of βHOB supplied.

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  Figure 5. Maximal and glucose mediated proton efflux rate from CO2 (PERCO2) increased with increasing concentrations of βHOB in both control and combination exposed cardiomyocytes. (A) Maximal PERCO2, (B) Maximal PER from lactate (PERLactate), (C) Glucose mediated PERCO2, and (D) Glucose mediated PERLactate in control and combination exposed NRCM. Bars represent mean PER (n = 10–11 litter per group). Blue color indicates controls and red color indicates combination exposed. Open symbols in the graph indicate males and filled symbols indicate females. Ketone effect was determined by two-way ANOVA with p values above straight bars; group differences as determined by one-way ANOVA are shown with p values above footed bars. Significance accepted at p < 0.05. mM refers to the progressive concentration of βHOB supplied.

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  Figure 6. The ratio of glucose mediated PERCO2 and PERLactate, a marker of aerobic glycolysis, increased with increasing concentration of βHOB in both control and combination exposed cardiomyocytes. (A) Maximal and (B) glucose-mediated PERCO2:PERLactate ratio for control and combination exposed NRCM. Bars represent mean ratio (n = 10–11 litter per group). Blue color indicate controls and red color indicate combination exposed. Open symbols in the graph indicate males and filled symbols indicate females. Ketone effect was determined by two-way ANOVA and the significance accepted at p < 0.05. (C) Bar graphs illustrate the combined contributions of PERCO2 (black with red border) and PERLactate (grey with blue border) to the PERTotal in control and combination exposed NRCM. Bars represent the mean ± SD. For illustration purposes, the solid line highlights the mean PERCO2 and the dashed line highlights the mean PERLactate for control NRCM that were not administered βHOB. mM refers to the progressive concentration of βHOB supplied.

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  Figure 7. Expression of genes involved in ketone body metabolism increase in combination exposed males. (A) Expression of Hmgcs2 and (B) Bdh1 in the neonatal heart of control and combination groups. Bars represents mean fold change (n = 12). Blue color indicate controls and red color indicate combination exposed. Bars represent mean fold change and the significance accepted at p < 0.05.

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  Figure 8. Maternal diabetes and HFD augment the myocardial expression of Pgc1a and Pparg. (A) Expression of Pgc1a, and (B) Pparg in the neonatal heart of control and combination groups. Bars represent mean fold change (n = 12). Blue color indicates controls and red color indicates combination exposed. Bars represent mean fold change and the significance accepted at p < 0.05.

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  Figure 9. Combination exposed males have increased myocardial expression of Cpt1a. Bars represents mean fold change (n = 12). Blue color indicates controls and red color indicates combination exposed. Bars represent mean fold change and the significance accepted at p < 0.05.

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  Figure 10. Schematic representation of the ketone stress test (KST) for assessing mitochondrial respiration in terms of oxygen consumption rate (OCR).