Maternal Exercise Programs Glucose and Lipid Metabolism and Modulates Hepatic miRNAs in Adult Male Offspring

Abstract

Detrimental exposures in mothers are recognized as risk factors for the development of metabolic dysfunction in offspring. In contrast, maternal exercise has been reported to be an effective strategy to maintain offspring health. However, the mechanisms underlying the protective effects of maternal exercise on adult offspring metabolic homeostasis are largely unclear. This study aims to investigate whether maternal exercise before and during pregnancy could combat the adverse effects of maternal high-fat diet (HFD) on metabolism in 24-week-old male offspring and to explore the role of miRNAs in mediating the effects. Female C57BL/6 mice were fed with either control diet or HFD 3-week prior to breeding and throughout pregnancy and lactation, among whom half of the HFD-fed mice were submitted to voluntary wheel running training 3-week before and during pregnancy. Male offspring were sedentary and fed with a control diet from weaning to 24 weeks. Body weight, the content of inguinal subcutaneous adipose tissue and perirenal visceral adipose tissue, glucose tolerance, and serum insulin and lipids in offspring were analyzed. Hepatic tissues were collected for transcriptome and miRNA sequencing and reverse transcription-quantitative polymerase chain reaction validation. The results showed that maternal HFD resulted in significant glucose intolerance, insulin resistance, and dyslipidemia in adult offspring, which were negated by maternal exercise. Transcriptome sequencing showed that maternal exercise reversed perinatal HFD-regulated genes in adult offspring, which were enriched in glucose and lipid metabolic-related signaling pathways. At the same time, maternal exercise significantly rescued the changes in the expression levels of 3 hepatic miRNAs in adult offspring, and their target genes were involved in the regulation of cholesterol biosynthesis and epigenetic modification, which may play an important role in mediating the intergenerational metabolic regulation of exercise. Overall, our research pioneered the role of miRNAs in mediating the programming effects of maternal exercise on adult offspring metabolism, which might provide novel insight into the prevention and treatment of metabolic disorders in early life.

Keywords: glucose and lipid metabolism; maternal voluntary wheel running training; miRNAs; mice; offspring; transcriptome.

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Copyright © 2022 Zhou, Li, Zhang, Yu and Xiao.

Figures

Figure 1

Glucose and lipid metabolism in male offspring. (A) Experimental scheme; (B) body weight changes; (C) subcutaneous adipose tissue mass at 24 weeks; (D) visceral adipose tissue content at 24 weeks; (E) AUC of blood glucose values during intraperitoneal glucose tolerance tests; (F) intraperitoneal glucose tolerance test at 24 weeks. C, offspring of dams fed the normal control diet; HF, offspring of dams fed the high-fat diet; HF-EX, offspring of dams intervened with a high-fat diet and exercise; SAT, subcutaneous adipose tissue mass; VAT, visceral adipose tissue; IPGTT, intraperitoneal glucose tolerance test; AUC, area under the curve. Data are expressed as means ± SD (n = 6/group) and are analyzed by one-way ANOVA or two-way ANOVA, with Turkey post-hoc analyses. Mean values were significantly different between the groups: *HF vs. C, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; #HF-EX vs. HF, #p < 0.05, ##p < 0.01, ###p < 0.001.

Figure 2

Principal component analysis and scatter plot of hepatic transcriptome sequencing in adult offspring. (A) Principal component analysis; (B) scatter plot of HF vs. C; (C) scatter plot of HF-EX vs. HF. C, offspring of dams fed the normal control diet; HF, offspring of dams fed the high-fat diet; HF-EX, offspring of dams intervened with a high-fat diet and exercise (n = 3/group).

Figure 3

Hepatic genes that were up-regulated by maternal HFD and down-regulated by maternal exercise and their functional analysis in adult offspring. (A) Heatmap of the differentially expressed genes; (B) Venn diagram of the differentially expressed genes; (C) Reactome pathway enrichment of the differentially expressed genes. C, offspring of dams fed the normal control diet; HF, offspring of dams fed the high-fat diet; HF-EX, offspring of dams intervened with a high-fat diet and exercise (n = 3/group); HFD, high-fat diet.

Figure 4

Hepatic genes that were down-regulated by maternal HFD and up-regulated by maternal exercise and their functional analysis in adult offspring. (A) Heatmap of the differentially expressed genes; (B) Venn diagram of the differentially expressed genes; (C) KEGG pathway enrichment of the differentially expressed genes; (D) differentially expressed genes. C, offspring of dams fed the normal control diet; HF, offspring of dams fed the high-fat diet; HF-EX, offspring of dams intervened with a high-fat diet and exercise; HFD, high-fat diet. Data are expressed as means ± SD (n = 3/group). Mean values were significantly different between the groups: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Figure 5

Maternal HFD and exercise regulated hepatic miRNA expression in adult offspring. (A) Volcano plot of miRNAs between HF and C groups; (B) volcano plot of miRNAs between HF-EX and HF groups; (C) Venn diagram of the differentially expressed miRNAs among the three groups; (D) lists of the three differentially expressed miRNAs. C, offspring of dams fed the normal control diet; HF, offspring of dams fed the high-fat diet; HF-EX, offspring of dams intervened with a high-fat diet and exercise; HFD, high-fat diet. Data are expressed as means ± SD (n = 3 / group). Mean values were significantly different between the groups: *p < 0.05, **p < 0.01.

Figure 6

Analysis of the targeted genes of the differentially expressed hepatic miRNAs in adult offspring regulated by maternal exercise. (A) Reactome pathway enrichment analysis of the target genes; (B) genes enriched in the epigenetic regulation of gene expression pathway; and (C) target genes involved in the regulation of cholesterol biosynthesis by SREBP pathway; (D)SREBF1 expression level. C, offspring of dams fed the normal control diet; HF, offspring of dams fed the high-fat diet; HF-EX, offspring of dams intervened with a high-fat diet and exercise; SREBF1, sterol regulatory element binding protein-1. Data are expressed as means ± SD (n = 3/group). Mean values were significantly different between the groups: **p < 0.01, ****p < 0.0001.