Influence of Maternal Metabolic Status and Diet during the Perinatal Period on the Metabolic Programming by Leptin Ingested during the Suckling Period in Rats

Abstract

We aimed to analyze the long-term metabolic effects of leptin supplementation at physiological doses during suckling in the offspring of diet-induced obese rats, together with the potential benefits of improving maternal diet during lactation. Thus, the offspring of: dams fed standard-diet (SD) (CON-dams), dams fed western-diet (WD) before and during gestation and lactation (WD-dams), and dams fed as WD-dams but moved to SD during lactation (REV-dams) were supplemented throughout suckling with leptin or vehicle, and fed SD or WD from weaning to four months. Under SD, leptin treatment significantly improved metabolic profile and body fat accumulation, with stronger effects in the male offspring of CON-dams and REV-dams. Under WD, the offspring of WD-dams presented metabolic alterations that were not evident in the offspring of REV-dams. Moreover, leptin supplementation improved glucose homeostasis in the male offspring of REV-dams. Conversely, leptin supplementation in females born to WD-dams and fed WD from weaning resulted in impaired insulin sensitivity and increased hepatic lipid content. These results highlight the importance of a balanced maternal diet during the perinatal period, especially lactation, for the subsequent metabolic health of the offspring and for the beneficial effects of leptin supplementation during suckling, more evident in the male offspring.

Keywords: Western diet; lactation; leptin; metabolic programming.

Figure 1

Representative scheme of the experimental design. Abbreviations: control dams (CON-dams), western diet dams (WD-dams), reversion dams (REV-dams), offspring of CON-dams (O-CON), offspring of WD-dams (O-WD), offspring of REV-dams (O-REV), vehicle (V), leptin (L), retroperitoneal white adipose tissue (rWAT).

Figure 2

Body weight and composition (fat and lean mass percentage) of O-CON, O-WD and O-REV male and female animals treated with vehicle or leptin during suckling and fed SD or WD from weaning, at one and four months of age. Data are mean ± SEM (n = 8–12). Statistics: After data separation depending on post-weaning diet, three-way ANOVA was performed to analyze the effect of sex, maternal diet and/or leptin treatment. In each sex, two-way ANOVA was performed to analyze the effect of leptin treatment and/or maternal diet. When an interactive effect was found, one-way ANOVA was performed to analyze the effect of maternal diet in each treatment. Single comparisons between leptin versus vehicle-treated rats of all experimental groups were carried out using Mann-Whitney U test. Symbols: sex (S), maternal diet (MD), leptin treatment (L); A ≠ B ≠ C (p < 0.05, LSD post-hoc, two-way ANOVA); a ≠ b (p < 0.05, LSD post-hoc, one-way ANOVA); *, different from their vehicle-treated equal (p < 0.05, Mann-Whitney U test). Abbreviations: offspring of CON-dams (O-CON), offspring of WD-dams (O-WD), offspring of REV-dams (O-REV), vehicle (V), leptin (L).

Figure 3

Cumulative food intake from one to four months of age of O-CON, O-WD and O-REV male and female animals treated with vehicle or leptin during suckling and fed SD or WD from weaning. Data are mean ± SEM (n = 8–12). Statistics: After data separation depending on post-weaning diet, three-way ANOVA was performed to analyze the effect of sex, maternal diet and/or leptin treatment. In each sex, two-way ANOVA was performed to analyze the effect of leptin treatment and/or maternal diet. Single comparisons between leptin versus vehicle-treated rats of all experimental groups were carried out using Mann-Whitney U test. Symbols: sex (S), maternal diet (MD), leptin treatment (L); A ≠ B (p < 0.05, LSD post-hoc, two-way ANOVA); *, different from their vehicle-treated equal (p < 0.05, Mann-Whitney U test). Abbreviations: offspring of CON-dams (O-CON), offspring of WD-dams (O-WD), offspring of REV-dams (O-REV), vehicle (V), leptin (L).

Figure 4

Circulating parameters and HOMA-IR of O-CON, O-WD and O-REV male and female animals treated with vehicle or leptin during suckling and fed SD or WD from weaning, under fasting (at three and half months) and ad libitum (four months) feeding conditions. Data are mean ± SEM (n = 8–12). Statistics: After data separation depending on post-weaning diet, three-way ANOVA was performed to analyze the effect of sex, maternal diet and/or leptin treatment. In each sex, two-way ANOVA was performed to analyze the effect of leptin treatment and/or maternal diet. Single comparisons between leptin versus vehicle-treated rats of all experimental groups were carried out using Mann-Whitney U test. Symbols: sex (S), maternal diet (MD), leptin treatment (L); A ≠ B (p < 0.05, LSD post-hoc, two-way ANOVA); *, different from their vehicle-treated equal (p < 0.05, Mann-Whitney U test). Abbreviations: offspring of CON-dams (O-CON), offspring of WD-dams (O-WD), offspring of REV-dams (O-REV), vehicle (V), leptin (L).

Figure 5

Hepatic lipid content in O-CON, O-WD and O-REV male and female animals treated with vehicle or leptin during suckling and fed SD or WD from weaning, at four months of age. Data are mean ± SEM (n = 8–12). Statistics: After data separation depending on post-weaning diet, three-way ANOVA was performed to analyze the effect of sex, maternal diet and/or leptin treatment. In each sex, two-way ANOVA was performed to analyze the effect of leptin treatment and/or maternal diet. When an interactive effect was found, one-way ANOVA was performed to analyze the effect of maternal diet in each treatment. Single comparisons between leptin versus vehicle-treated rats of all experimental groups were carried out using Mann-Whitney U test. Symbols: sex (S), maternal diet (MD), leptin treatment (L); a ≠ b (p < 0.05, LSD post-hoc, one-way ANOVA); *, different from their vehicle-treated equal (p < 0.05, Mann-Whitney U test). Abbreviations: offspring of CON-dams (O-CON), offspring of WD-dams (O-WD), offspring of REV-dams (O-REV), vehicle (V), leptin (L).

Figure 6

Expression levels of genes related to energy metabolism in retroperitoneal white adipose tissue of O-CON, O-WD and O-REV male and female animals treated with vehicle or leptin during suckling and fed SD or WD from weaning, at four months of age. Data are mean ± SEM (n = 8–12). Statistics: After data separation depending on post-weaning diet, three-way ANOVA was performed to analyze the effect of sex, maternal diet and/or leptin treatment. In each sex, two-way ANOVA was performed to analyze the effect of leptin treatment and/or maternal diet. When an interactive effect was found, one-way ANOVA was performed to analyze the effect of maternal diet in each treatment. Single comparisons between leptin versus vehicle-treated rats of all experimental groups were carried out using Mann-Whitney U test. Symbols: sex (S), maternal diet (MD), leptin treatment (L); A ≠ B (p < 0.05, LSD post-hoc, two-way ANOVA); a ≠ b (p < 0.05, LSD post-hoc, one-way ANOVA); *, different from their vehicle-treated equal (p < 0.05, Mann-Whitney U test). Abbreviations: offspring of CON-dams (O-CON), offspring of WD-dams (O-WD), offspring of REV-dams (O-REV), vehicle (V), leptin (L).

Figure 7

Expression levels of genes related to energy metabolism in liver of O-CON, O-WD and O-REV male and female animals treated with vehicle or leptin during suckling and fed SD or WD from weaning, at four months of age. Data are mean ± SEM (n = 8–12). Statistics: After data separation depending on post-weaning diet, three-way ANOVA was performed to analyze the effect of sex, maternal diet and/or leptin treatment. In each sex, two-way ANOVA was performed to analyze the effect of leptin treatment and/or maternal diet. When an interactive effect was found, one-way ANOVA was performed to analyze the effect of maternal diet in each treatment. Single comparisons between leptin versus vehicle-treated rats of all experimental groups were carried out using Mann-Whitney U test. Symbols: sex (S), maternal diet (MD), leptin treatment (L); A ≠ B (p < 0.05, LSD post-hoc, two-way ANOVA); a ≠ b (p < 0.05, LSD post-hoc, one-way ANOVA); *, different from their vehicle-treated equal (p < 0.05, Mann-Whitney U test). Abbreviations: offspring of CON-dams (O-CON), offspring of WD-dams (O-WD), offspring of REV-dams (O-REV), vehicle (V), leptin (L).

Figure 8

Summary of the effects of leptin supplementation during suckling on cumulative food intake (from one to four months of age), fat mass and hepatic lipid content (four months), and circulating parameters in adult O-CON, O-WD and O-REV male and female rats weaned onto SD or WD. Circulating parameter were determined at four months (under feeding conditions), or at three and a half months (under fasting conditions). Abbreviations: offspring of CON-dams (O-CON), offspring of WD-dams (O-WD), offspring of REV-dams (O-REV), standard diet (SD), western diet (WD). Symbols: ↑ and ↓ indicate increases or decreases, respectively, by Mann-Whitney U test, two-way ANOVA, or three-way ANOVA with respect to their vehicle-treated equals; ●, significant difference in the indicated group with respect to its vehicle-treated equal by Mann-Whitney U test.