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).