Alternate-day fasting delays pubertal development in normal-weight mice but prevents high-fat diet-induced obesity and precocious puberty

Abstract

Background/objectives: Childhood obesity, particularly in girls, is linked to early puberty onset, heightening risks for adult-onset diseases. Addressing childhood obesity and precocious puberty is vital to mitigate societal burdens. Despite existing costly and invasive medical interventions, introducing lifestyle-based alternatives is essential. Our study investigates alternate-day fasting's (ADF) impact on pubertal development in normal-weight and high-fat diet (HFD)-induced obese female mice.

Methods: Four groups of female mice were utilized, with dams initially fed control chow during and before pregnancy. Post-parturition, two groups continued on control chow, while two switched to an HFD. Offspring diets mirrored maternal exposure. One control and one HFD group were subjected to ADF. Morphometry and hormone analyses at various time points were performed.

Results: Our findings demonstrate that ADF in normal-weight mice led to reduced body length, weight, uterine, and ovarian weights, accompanied by delayed puberty and lower levels of sex hormones and growth hormone (GH). Remarkably, GH treatment effectively prevented ADF-induced growth reduction but did not prevent delayed puberty. Conversely, an HFD increased body length, induced obesity and precocious puberty, and altered sex hormones and leptin levels, which were counteracted by ADF regimen. Our data indicate ADF's potential in managing childhood obesity and precocious puberty.

Conclusions: ADF reduced GH and sex hormone levels, contributing to reduced growth and delayed puberty, respectively. Therefore, parents of normal-weight children should be cautious about prolonged overnight fasting. ADF prevented HFD-induced obesity and precocious puberty, offering an alternative to medical approaches; nevertheless, further studies are needed for translation into clinical practice.

Fig. 1. Effects of 18 h ADF on pubertal development in normal weight mice.

a Cumulative percentage of VO; b Average age at puberty onset (indicated by VO); c–e Body weight at P34 and P40; f–h Body length at P34 and P40; i–k Uterine weight at P34 and P40; l–n Ovarian weight at P34 and P40. Data are presented as mean ± SEM, and p-values were calculated using the unpaired Student’s t test and the Mann–Whitney test. Each data point corresponds to an individual mouse. PND Postnatal days. The data represent three separate experiments.

Fig. 2. Effects of 18 h ADF on plasma levels of sex hormones in normal weight mice.

a–c Plasma levels of LH at P34 and P40; d–f Plasma levels of FSH at P34 and P40; g–i Plasma levels of E2 at P34 and P40. Data are presented as mean ± SEM, and p-values were calculated using the unpaired Student’s t test and the Mann–Whitney test. Each data point corresponds to an individual mouse. The data represent two separate experiments.

Fig. 3. Effects of 18 h ADF on plasma levels of metabolic hormones and growth factors in normal weight mice.

a–c Plasma levels of leptin at P34 and P40; d–f Plasma levels of insulin at P34 and P40; g–i Plasma levels of GH at P34 and P40; j–l Plasma levels of IGF-1 at P34 and P40. Data are presented as mean ± SEM, and p-values were calculated using the unpaired Student’s t test and the Mann–Whitney test. Each data point corresponds to an individual mouse. The data represent two separate experiments.

Fig. 4. Effects of 18 h ADF on pubertal development in HFD-induced obese mice.

a Cumulative percentage of VO; b Average age at puberty onset; c Body weight at P21; d Body weight at P34; e Body length at P34; f Uterine weight at P34; g Ovarian weight at P34; h Plasma levels of LH at P34; i Plasma levels of FSH at P34; j Plasma levels of E2 at P34; k Plasma levels of GH at P34; l Plasma levels of IGF-1 at P34. Data are presented as mean ± SEM, and p-values were calculated using one-way ANOVA and the Kruskal–Wallis test. Each data point corresponds to an individual mouse. PND Postnatal days. The data represent two separate experiments.

Fig. 5. Effects of 24 h ADF on pubertal development in HFD-induced obese mice.

a Cumulative percentage of VO; b Average age at puberty onset; c Body weight at P34; d Body length at P34; e Uterine weight at P34; f Ovarian weight at P34; g Plasma levels of LH at P34; h Plasma levels of FSH at P34; i Plasma levels of E2 at P34; j Plasma levels of GH at P34; k Plasma levels of IGF-1 at P34; l Plasma levels of insulin at P34; m Plasma levels of leptin at P34. Data are presented as mean ± SEM, and p-values were calculated using the one-way ANOVA and the Kruskal–Wallis test. Due to differences in variance, the Kruskal–Wallis test did not indicate a significant difference in uterine weight between the Cntrl and HFD groups, but the Mann–Whitney test did (Supplementary Fig. 2). Each data point corresponds to an individual mouse. PND Postnatal days. The data represent three separate experiments.

Fig. 6. GH administration prevents reduced body growth induced by 18 h ADF.

a Cumulative percentage of VO; b Average age at puberty onset; c Body weight at P40; d Body length at P40; e Uterine weight at P40; f Ovarian weight at P40. Data are presented as mean ± SEM, and p-values were calculated using the Kruskal–Wallis test. Due to variance differences, the Kruskal–Wallis test did not indicate significance for nose-to-anus length, uterine weight, and ovarian weight between the Cntrl and HFD groups. However, the Mann–Whitney and unpaired t-tests did show significance (Supplementary Fig. 3). Each data point corresponds to an individual mouse. PND Postnatal days. The data represent three separate experiments.