Voluntary Wheel Running Reduces Cardiometabolic Risks in Female Offspring Exposed to Lifelong High-Fat, High-Sucrose Diet

Abstract

Purpose: Maternal and postnatal overnutrition has been linked to an increased risk of cardiometabolic diseases in offspring. This study investigated the impact of adult-onset voluntary wheel running to counteract cardiometabolic risks in female offspring exposed to a life-long high-fat, high-sucrose (HFHS) diet.

Methods: Dams were fed either an HFHS or a low-fat, low-sucrose (LFLS) diet starting from 8 wk before pregnancy and continuing throughout gestation and lactation. Offspring followed their mothers' diets. At 15 wk of age, they were divided into sedentary (Sed) or voluntary wheel running (Ex) groups, resulting in four groups: LFLS/Sed ( n = 10), LFLS/Ex ( n = 5), HFHS/Sed ( n = 6), HFHS/Ex ( n = 5). Cardiac function was assessed at 25 wk, with tissue collection at 26 wk for mitochondrial respiratory function and protein analysis. Data were analyzed using two-way ANOVA.

Results: Although maternal HFHS diet did not affect the offspring's body weight at weaning, continuous HFHS feeding postweaning resulted in increased body weight and adiposity, irrespective of the exercise regimen. HFHS/Sed offspring showed increased left ventricular wall thickness and elevated expression of enzymes involved in fatty acid transport (CD36, FABP3), lipogenesis (DGAT), glucose transport (GLUT4), oxidative stress (protein carbonyls, nitrotyrosine), and early senescence markers (p16, p21). Their cardiac mitochondria displayed lower oxidative phosphorylation (OXPHOS) efficiency and reduced expression of OXPHOS complexes and fatty acid metabolism enzymes (ACSL5, CPT1B). However, HFHS/Ex offspring mitigated these effects, aligning more with LFLS/Sed offspring.

Conclusions: Adult-onset voluntary wheel running effectively counteracts the detrimental cardiac effects of a lifelong HFHS diet, improving mitochondrial efficiency, reducing oxidative stress, and preventing early senescence. This underscores the significant role of physical activity in mitigating diet-induced cardiometabolic risks.

Figure 1.

Voluntary wheel running and physiological measurements. A. Daily running distance (Km/Day); B. Daily running duration (Hr/Day); C. Sol wt/BW; D. Final BW; E. HW; F. LVW; G. Visceral fat; H. Sub fat for studied animals. Values are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. n= 10 in LFLS/Sed; n= 6 in HFHS/Sed, n= 5 in LFLS/Ex; n= 5 in HFHS/Ex. Sed: sedentary; Ex: voluntary wheel running; LFLS: low-fat and low-sucrose diet; HFHS: high-fat and high-sucrose diet; Km: kilometer; Hr: hours; Sol wt: soleus weight; BW: body weight; HW: heart weight; LVW: Left ventricular weight; Sub: subcutaneous.​

Figure 2.

Voluntary wheel running relieves HFHS-induced metabolic maladaptation. A. Representative Western blot images of lipolysis-related proteins. B-D. Quantification of lipolysis signaling proteins ATGL, ABHD5, p-HSL, HSL, and p-HSL/HSL. E. Representative Western blot images. F-J. Quantification of fatty acid import (CD36 and FABP3), activation (ACSL1), and lipogenesis (FASN and DGAT1) proteins. K. Representative Western blot images of insulin signaling-related proteins and glucose transportation proteins. L-P. Quantification of insulin signaling activation markers p-IRβ, IRβ, p-AKT, AKT, p-mTOR, mTOR, AS160, and GLUT4. Values are mean ± SEM. * P < 0.05, ** P < 0.01. 4 animals per group with 3 technical replicates per animal were used. Ponceau staining was used as a loading control. Sed: sedentary; Ex: voluntary wheel running; LFLS: low-fat and low-sucrose diet; HFHS: high-fat and high-sucrose diet; ATGL: adipose triglyceride lipase; ABHD5: alpha-beta hydrolase domain-containing 5; HSL: hormone-sensitive lipase; CD36: Cluster of differentiation; FABP3: fatty acid binding protein 3; ACSL1: Acyl-CoA synthetase long-chain family member 1; FASN: fatty acid synthetase; DGAT1: diacylglycerol O-acyltransferase 1; IRb: insulin receptor β; mTOR: mammalian target of rapamycin; AS160: AKT substrate 160; GLUT4: glucose transport 4; p: phosphorylated form.

Figure 3.

Mitochondrial respiration measured by high-resolution respirometry. A. Quantification of CS activities in the isolated mitochondria, normalized to mg of isolated mitochondrial protein. B-E. OXPHOS capacity in the absence of fatty acids substrates. B. PM_L: Leak respiration in the presence of pyruvate and malate (PM); C. PMp: PM-supported OXPHOS-linked respiration; D. CI+CIIp: maximal OXPHOS rates; E. (PMp-PML)/PMp: coupling efficiency. F-J. OXPHOS capacity in the presence of fatty acids substrates, Palmitoyl-L-carnitine + Malate (PalM). F. FAO_L: leak respiration; G. FAOp: ADP-stimulated OXPHOS; H. FAOp CIp: PalM-supported OXPHOS; I. FAOp CI+CIIp: Maximal OXPHOS-linked respiration. J. (FAOp-FAO_L)/FAOp: Coupling efficiency. Values are expressed as mean ± SEM. n= 10 in LFLS; n= 6 in HFHS, n= 5 in LFLS/Ex; n= 5 in HFHS/Ex. ** P < 0.01. Sed: sedentary; Ex: voluntary wheel running; LFLS: low-fat and low-sucrose diet; HFHS: high-fat and high-sucrose diet; CS: Citrate synthase; CI: complex I; CII: complex II; FAO: fatty acid oxidation; JO₂: oxygen flux.

Figure 4.

Voluntary wheel running reinforces metabolic efficiency. A. Representative Western blot images from isolated mitochondrial fractions B. Quantification of mitochondrial electron transport chain proteins. C. Representative Western blot images. D-F. Quantification of fatty acid activation at mitochondria (ACSL5), fatty acid transportation to mitochondria (CPT1B and CPT2). Ponceau staining was used as a loading control. Values are mean ± SEM. 4 animals per group with 3 technical replicates per animal were used. * P < 0.05. Sed: sedentary; Ex: voluntary wheel running; LFLS: low-fat and low-sucrose diet; HFHS: high-fat and high-sucrose diet; NDUFB8: NADH: ubiquinone oxidoreductase subunit B8; SDHB: Succinate dehydrogenase subunit B; UQCRC2: Ubiquinol-cytochrome c reductase core protein 2; MTCO1: Mitochondrially encoded cytochrome c oxidase I; ATP5F1A: ATP synthase F1 subunit alpha; ACSL5: Acyl-CoA synthetase long-chain family member 5; CPT1B: Carnitine palmitoyl transferase 1B; CPT2: Carnitine palmitoyl transferase 2.

Figure 5.

Voluntary wheel running reverts oxidative stress caused by protein modifications. A-H. Representative Western blot images and results obtained from whole tissue lysates. B. Quantification of antioxidant enzyme CuZnSOD; C-D. Quantification of lipid peroxidation makers 4-HNE and MDA. E-F. Quantification of protein carbonylation with Representative Western blot images. G-H. Representative Western blot images with Quantification of nitrotyrosine; I-M. Representative Western blot images and results obtained from isolated mitochondria fractions; I. Representative Western blot images of 4-HNE; J. Quantification of lipid peroxidation maker, 4-HNE. K. Representative Western blot images of MAO-A and nitrotyrosine; L. Quantification of MAO-A; and M. nitrotyrosine, respectively. Values are mean ± SEM. 4 animals per group with 3 technical replicates per animal were used. Ponceau staining was used as a loading control. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Sed: sedentary; Ex: voluntary wheel running; LFLS: low-fat and low-sucrose diet; HFHS: high-fat and high-sucrose diet; CuZnSOD: Copper Zinc superoxide dismutase; 4HNE: 4-Hydroxy-2-nonenal; MDA: Malondialdehyde; MAO-A: monoamine oxidase A.

Figure 6.

Voluntary wheel running represses HFHS-diet-induced cardiac senescence. A. Representative Western blot images. B-D. Quantification of senescence-related proteins p53, p16, and p21, respectively. Values are mean ± SEM. 4 animals per group with 3 technical replicates per animal were used. Ponceau staining was used as a loading control. * P < 0.05, ** P < 0.01. Sed: sedentary; Ex: voluntary wheel running; LFLS: low-fat and low-sucrose diet; HFHS: high-fat and high-sucrose diet.