Figure 1 . Experimental Design. The mothers were divided into
four experimental groups during the pre-pregnancy period, until week 9:
G1- Water: females that received water and did not practice voluntary
physical exercise (n=10); G2 – Water + VPE: females that received water
and practiced voluntary physical exercise (n=20); G3 –FRU: females that
received 20% fructose/L (n=10); G4 –FRU + VPE: females that received
20% fructose/L and practiced voluntary physical exercise (n=30). After
this period, interventions were carried out for which the females were
subdivided into 7 groups: G1 - Water: females continued to receive water
and did not practice voluntary physical exercise until the end of
pregnancy (n=10); G2a – Water + VPE: females that received water and
practiced voluntary physical exercise until the end of pregnancy (n=10);
G2b – Water + VPE/Water: females that received water and practiced
voluntary physical exercise until copulation and then stopped the VPE
and continued only with water until the end of the pregnancy period
(n=10); G3 – FRU: females that continued to receive 20% fructose/L
(n=10); G4a – FRU + VPW: females that received 20% fructose/L and
practiced voluntary physical exercise until the end of pregnancy (n=10);
G4b –FRU + VPE/Water + VPE: females that received 20% fructose/L and
practiced voluntary physical exercise until copulation, after this
period stopped fructose consumption and continued receiving only water
and practiced voluntary physical exercise during pregnancy (n=10); G4c
– FRU + VPE/Water: females that received 20% fructose/L and practiced
voluntary physical exercise until copulation, after which they stopped
fructose consumption and voluntary physical exercise and received only
water during pregnancy (n=10). *During lactation only fructose
supplementation was continued, while VPE was stopped due to limited
space in the cages of the mothers and the pups. After weaning (week 15),
the offspring continued to be monitored according to the seven
experimental groups, referred to as offspring groups (OG). FRU:
fructose; VPE: Voluntary Physical Exercise;
blood collection from mother;
assessment of VPE;
blood collection from offspring.
Figure 2. Caloric intake (kcal) of mothers during the various
stages, while or after receiving water or fructose (FRU) and/or
practiced voluntary physical exercise (VPE) during pre-pregnancy and/or
pregnancy. Data are expressed as mean ± standard deviation of mean
consumption of calories (n=10 animals per group).aSignificant difference in relation to the water group
(p < 0.05, Kruskal-Wallis, post hoc Dunn’s).
Figure 3 . Fructose concentration (mg/mL) determined by high
performance liquid chromatography in the serum of mothers that received
water or fructose (FRU) and/or practiced voluntary physical exercise
(VPE) during pre-pregnancy and/or
pregnancy. Data are expressed as mean
± standard deviation of mean (n=6 animals per group).aSignificant difference in relation to the water group
(p < 0.05, Test t -student); bSignificant difference in relation to the fructose group (p <
0.05, Test t -student).
Figure 4. Insulin tolerance test of mothers that received water
or fructose (FRU) and/or practiced voluntary physical exercise (VPE)
during pre-pregnancy and/or pregnancy. Data are expressed as mean ±
standard deviation of mean (n=6 animals per group).aSignificant difference in relation to the water group
(p < 0.05, ANOVA, post hoc Tukey’s); bSignificant difference in relation to the fructose group (p <
0.05, ANOVA, post hoc Tukey’s). AUC: Area under the curve. Based
on the graphs on the left, the areas under the curves were computed and
presented in the graphs on the right in the insulin tolerance test.
Figure 5. DNA damage index in peripheral blood cells of mothers
that received water or fructose (FRU) and/or practiced voluntary
physical exercise (VPE) during pre-pregnancy and/or pregnancy. Data are
expressed as mean ± standard deviation of the mean (n=8 animals per
group). 5A: peripheral blood in pre-pregnancy; 5B: peripheral blood in
pregnancy; 5C: peripheral blood in post-pregnancy.aSignificant difference in relation to the water group
(p < 0.05, ANOVA, post hoc Tukey’s);bSignificant difference in relation to the fructose
group (p < 0.05, ANOVA, post hoc Tukey’s). FPG:
formamidopyrimidine DNA glycosylase.
Figure 6. DNA damage index (assessed as strand breaks) in liver
cells of mothers that received water or fructose (FRU) and/or practiced
voluntary physical exercise (VPE) during pre-pregnancy and/or pregnancy.
Data are expressed as mean ± standard deviation of the mean (n=8 animals
per group). aSignificant difference in relation to the
water group (p < 0.05, ANOVA, post hoc Tukey’s);bSignificant difference in relation to the fructose
group (p < 0.05, ANOVA, post hoc Tukey’s).
Figure 7 . Fructose concentration (mg/mL) determined by high
performance liquid chromatography of the offspring (females and males)
of mothers that received water or fructose and/or practiced voluntary
physical exercise during the pre-pregnancy period /or pregnancy. Data
are expressed as mean ± standard deviation of mean (n=6 animals per
group). FRU- Fructose; VPE- voluntary physical exercise.aSignificant difference in relation to the water group
(p < 0.05, ANOVA, post hoc Tukey’s);bSignificant difference in relation to the FRU group
(p < 0.05, ANOVA, post hoc Tukey’s).
Figure 8. Body adiposity index of offspring (females and males)
of mothers that received water or fructose and/or practiced voluntary
physical exercise during pre-pregnancy and/or pregnancy. Data are
expressed as mean ± standard deviation of mean (n=6 animals per group).
FRU- Fructose; VPE- voluntary physical exercise.aSignificant difference in relation to the water group
(p < 0.05, ANOVA, post hoc de Tukey’s);bSignificant difference in relation to the FRU group
(p < 0.05, ANOVA, post hoc de Tukey’s).
Figure 9. Insulin tolerance test of offspring (females and
males) of mothers that received water or fructose and/or practiced
voluntary physical exercise during pre-pregnancy and/or pregnancy. Data
are expressed as mean ± standard deviation of mean (n=6 animals per
group). FRU- Fructose; VPE- voluntary physical exercise.aSignificant difference in relation to the FRU group
(p < 0.05, ANOVA, post hoc Tukey’s);bSignificant difference compared to the FRU + VPE
group (p < 0.05, ANOVA, post hoc Tukey’s). AUC: area
under the curve.
Figure 10. Histological profile of the hepatic tissue of the
offspring (females and males) of mothers that received water or fructose
and/or practiced voluntary physical exercise during pre-pregnancy and/or
pregnancy. The upper pannel shows the representative images of female
and male offspring. The lower pannel shows the quantified percentage of
steatosis of female and male offspring. Representative images of
histological photomicrographs of cross-sections of the liver of the
animals. A: Water; B: Water + VPE; C: Water + VPE/Water; D: FRU; E: FRU
+ VPE; F: FRU + VPE/Water + VPE; G: FRU + VPE/Water. Data are expressed
as mean ± standard deviation of mean (n=8 animals per group). FRU-
Fructose; VPE- voluntary physical exercise.aSignificant difference in relation to the water group
(p < 0.05, Kruskal-Wallis, post hoc Dunn’s);bSignificant difference in relation to the FRU group
(p < 0.05, Kruskal-Wallis, post hoc Dunn’s);cSignificant difference in relation to the FRU + VPE
group (p < 0.05, Kruskal-Wallis, post hoc Dunn’s);dSignificant difference compared to the FRU +
VPE/Water + VPE group (p < 0.05, Kruskal-Wallis, post
hoc Dunn’s).
Figure 11. DNA damage index in peripheral tissue cells of the
offspring (female and male) of mothers that received water or fructose
(FRU) and/or practiced voluntary physical exercise (VPE) during
pre-pregnancy and/or pregnancy. Data are expressed as mean ± standard
deviation of mean (n=8 animals per group).aSignificant difference in relation to the water group
(p < 0.05, ANOVA, post hoc Tukey’s);bSignificant difference in relation to the FRU group
(p < 0.05, ANOVA, post hoc Tukey’s).