Table 1.
Forward and reverse sequences of primers used for the amplification of mouse genes.
Figure 1.
Increase in body weight and body composition during life in mice postnatally overfed (OF) by litter size reduction (3 pups/litter) or normally fed (NF) in litters of normal size (10 pups/litter).
A: Body weight from birth to 210 days (7 months) of OF and NFmice. Results are expressed as means ± SEM. *, P<0.05 was considered significant (one-way ANOVA). B: Percentage of mean body weight increase in OF mice as compared with that in NF mice. C: Body composition (fat mass, lean mass and total water), measured by nuclear magnetic resonance, as a percentage of body weight in OF mice and NF mice. *, P<0.05 was considered significantly different. Data are expressed as means ± SEM from 10 animals per group.
Table 2.
Metabolic parameters of NF and OF mice groups. Fasting blood glucose was assayed by glucometry; cholesterol and triglycerides were assayed using colorimetric methods and plasma insulin, leptin and adiponectin were estimated using ELISA. P was calculated by Student’s t-test.
Table 3.
List of genes differentially expressed between NF and OF mice at 24 days of age.
Figure 2.
Apelin and apelin receptor (APJ) gene expression in myocardial tissue mRNA extracts from young (24 days) postnatally normal-fed (NF) or overfed (OF) mice.
The mRNA expression of (A) apelin and (B) apelin receptor genes was measured by quantitative RT-PCR in myocardial tissues extracted from juvenile (24 days) mice that were overfed (OF) by litter size reduction (3 pups/litter) or normally fed (NF) in litters of normal size (10 pups/litter). Results are expressed as means ± SEM from 8 animals per group. P was calculated by Student’s t-test.
Figure 3.
Oxidative stress indexes in plasma and heart tissue of 7-month-old mice that were postnatally overfed (OF) by litter size reduction (3 pups/litter) or normally fed (NF) in litters of normal size (10 pups/litter).
A: Plasma hydroperoxides, considered as a marker of peroxidation in the circulation, were measured in OF and NF mice (FORT test). B: Cardiac MnSOD mRNA relative expression measured by quantitative RT-PCR in myocardial tissues extracted from adult (7 months) NF or OF mice. C: Cardiac catalase mRNA relative expression measured by quantitative RT-PCR in myocardial tissues extracted from adult (7 months) NF or OF mice. D: Cardiac global superoxide dismutase (SOD) enzymatic activity. E: Cardiac catalase enzymatic activity. F: CP• nitroxide radical formation in heart tissue, measured with electron paramagnetic resonance spectroscopy. Under the presence of reactive nitrogen and oxygen species, the cell-permeable CMH hydroxylamine probe is converted into the nitroxide CP• radical, which can be considered a marker of tissue nitro-oxidative stress. G: Representative anisotropic signal of CP• radical in heart tissue at 100K. Data are expressed as means ± SEM from 10 animals per group. P was calculated by Student’s t-test.
Figure 4.
Cardiovascular function in anesthetized 7-month-old mice that were postnatally overfed (OF) by litter size reduction (3 pups/litter) or normally fed (NF) in litters of normal size (10 pups/litter).
A: Diastolic (DBP), systolic (SBP) and mean arterial blood pressure (MABP) measured at 7 months by plethysmography. B: Evolution of left ventricular ejection fraction (LVEF, in %) from 3 months to 7 months of life, measured in vivo by echocardiography. C: Evolution of left ventricular internal diameter (LVID) from 3 months to 7 months of life, measured in vivo by echocardiography. *, P<0.05; ***, P<0.001 were considered significantly different from NF mice. D: left ventricular ejection fraction measured at 7 months in anesthetized NF and OF mice by echocardiography (LVEF, in %). E: left ventricular end diastolic diameter (LVEDD) and left ventricular end systolic diameter (LVESD) measured at 7 months in anesthetized NF and OF mice by echocardiography. Data are expressed as means ± SEM from 10 animals per group. P was calculated by Student’s t-test.
Figure 5.
Ventricular remodeling of hearts from 7-month-old NF or OF mice.
A: Collagen density (% of area) of ventricular tissues from 7 months-old NF and OF mice. B: Representative picrosirius collagen staining of ventricular tissue from 7 months-old NF and OF mice., C: MMP-2 mRNA expression in myocardial tissues from 7 months-old NF and OF mice, measured by quantitative RT-PCR, D: Metalloproteinase activity in myocardial tissues from 7 months-old NF and OF mice, measured by gelatin zymography. P was calculated by Student’s t-test.
Figure 6.
Myocardial contractility recovery, tissue injury of hearts from 7-month-old NF or OF mice, after 30 min of global normothermic ischemia followed by 2 hours of reperfusion, and expression of STAT-3 and SOCS3 in intact (non-ischemic) ventricles of 7-months old NF or OF mice.
A: Recovery of pre-ischemic coronary flow (in %) of isolated Langendorff perfused hearts after 30 min of global normothermic total ischemia. B: Recovery of pre-ischemic developed pressure (in %) of isolated Langendorff perfused hearts after 30 min of global normothermic total ischemia. C: Ratios of ventricular infarcted areas/non-ischemic tissue (in %) of NF and OF mice hearts, after 30 min of global ischemia followed by 2 h of reperfusion. P was calculated by Student’s t-test. D: representative pictures of TTC-staining of myocardial slices of NF and OF mice after ischemia-reperfusion. Red-stained areas show non-ischemic tissue and white areas show infarcted tissue. E: Phosphorylated/total STAT-3 protein expression in intact ventricular tissue (non-ischemic) of NF and OF mice. F: SOCS-3/HSC70 protein expression measured by Western blot in intact ventricular tissue (non-ischemic) of NF and OF mice.