Abstract
Objective
Intriguingly, hyperinsulinemia, and hyperglycemia can predispose insulin resistance, obesity, and type 2 diabetes, leading to metabolic disturbances. Conversely, physical exercise stimulates skeletal muscle glucose uptake, improving whole-body glucose homeostasis. Therefore, we investigated the impact of short-term physical activity in a mouse model (Slc2a4+/−) that spontaneously develops hyperinsulinemia and hyperglycemia even when fed on a chow diet.
Methods
Slc2a4+/− mice were used, that performed 5 days of endurance or strength exercise training. Further analysis included physiological tests (GTT and ITT), skeletal muscle glucose uptake, skeletal muscle RNA-sequencing, mitochondrial function, and experiments with C2C12 cell line.
Results
When Slc2a4+/− mice were submitted to the endurance or strength training protocol, improvements were observed in the skeletal muscle glucose uptake and glucose metabolism, associated with broad transcriptomic modulation, that was, in part, related to mitochondrial adaptations. The endurance training, but not the strength protocol, was effective in improving skeletal muscle mitochondrial activity and unfolded protein response markers (UPRmt). Moreover, experiments with C2C12 cells indicated that insulin or glucose levels could contribute to these mitochondrial adaptations in skeletal muscle.
Conclusions
Both short-term exercise protocols were efficient in whole-body glucose homeostasis and insulin resistance. While endurance exercise plays an important role in transcriptome and mitochondrial activity, strength exercise mostly affects post-translational mechanisms and protein synthesis in skeletal muscle. Thus, the performance of both types of physical exercise proved to be a very effective way to mitigate the impacts of hyperglycemia and hyperinsulinemia in the Slc2a4+/− mouse model.
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Data availability
The data that support the findings of this study are available from the corresponding author upon reasonable request.
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Funding
This work was supported by FAEPEX, the National Council for Scientific and Technological Development (CNPq; case numbers 303571/2018–7; 140285/2016–4; 442542/2014–3 and 306535/2017–3), the Coordination for the Improvement of Higher Education Personnel (CAPES; finance code 001), and the São Paulo Research Foundation (FAPESP; case numbers 2015/26000–2, 2016/18488–8, 2018/20872–6, 2019/11820–5, 2020/13443–1 and 2021/08692–5).
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VRM and JRP wrote the paper and were ultimately responsible for the experiments in this study. VRM, JDB, RCG, ALR, RFLV, SCBRN and GCA. designed and performed the experiments with animals. BMC and RRB performed the mitochondrial function experiments. VRM, MBS, and FMS designed and performed the cell culture experiments. SQB, CDR, and LAV contributed to PET scan experiments. LAV, FMS, LPM, ASRS, ERR, DEC, and JRP contributed to the discussion and laboratory support. All the authors have read, critically revised, and approved this manuscript.
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18_2023_4771_MOESM1_ESM.tif
Figure S1. Morphological and physiological parameters. A. Fasting glucose at baseline (before exercise) and after (16 h and 90 h) exercise. B. Fasting insulin at baseline (before exercise) and after (16 h and 90 h) exercise. C. Body weight. D. Lee index. E. Total fat weight. F. Tissue weights (gastrocnemius, heart, mesenteric WAT, retroperitoneal WAT, perigonadal WAT, inguinal WAT, and brown adipose tissue). G. Representative images of hematoxylin–eosin (H&E, 10x, scale bar: 100 µM) in the liver. H. Liver tissue weight. I. Biochemical parameters in the serum (triglycerides, cholesterol, and high-density lipoprotein. J. mRNA levels of gluconeogenic genes (Pepck, G6pase, and Pc). In A-B, n = 7/group was used (* p < 0.05 vs WT group. # p < 0.05 vs Basal condition). In C-F,H n = 9/group was used. In I-J, n = 5/group was used. * p < 0.05 vs WT group. # p < 0.05 vs Slc2a4+/− Sedentary group. (TIF 13534 KB)
18_2023_4771_MOESM2_ESM.tif
Figure S2. Gene ontology analyses of up/downregulated genes in the skeletal muscle of Slc2a4+/− mice. A. Biological process, cellular components, and molecular functions of upregulated genes in the endurance versus sedentary comparison. B. Biological process, cellular components, and molecular functions of downregulated genes in the endurance versus sedentary comparison. C. Biological process, cellular components, and molecular functions of upregulated genes in the strength versus sedentary comparison. D. Biological process, cellular components, and molecular functions of downregulated genes in the strength versus sedentary comparison. E. Confirmation of common downregulated genes in the gastrocnemius of Slc2a4+/− mice (Cdh4, Ctgf1, Gas1, Hey1, Penk, Ptprb, Amd2, Sorbs2). F-G. Confirmation of common upregulated genes in the gastrocnemius of Slc2a4+/− mice (Srxn1, Irx3, Klhl40, Hspa1a, Hspa1b, Hspb1, Ninj1, Hspb7, Ogdha, Lmcd1, Nfic, Bcl2l13, Retsat, Hspa9, Dnaja4, Fads6, Hadha). In E–G, n = 5, 6, 6 was used. # p < 0.05 vs Slc2a4+/− Sedentary group. (TIF 10541 KB)
18_2023_4771_MOESM3_ESM.tif
Figure S3. Behavioral response after exercise protocols. A. Schematic figure of the groups submitted to the elevated plus maze and open field test (Slc2a4+/− Sedentary, and Slc2a4+/− Endurance or Strength after exercise protocol). B. Time in the border during the open field test. C. Time in the center during the open field test. D. Grooming times during the open field test. E. Percentage of time spent in the open arms during the elevated plus maze test. F. Percentage of time spent in the closed arms during the elevated plus maze test. G. Percentage of time spent in the center during the elevated plus maze test. H. Percentage of entries in the open arms during the elevated plus maze test. I. Corticosterone levels in the serum. In B-H, n = 5/group was used. In I, n = 4, 5, 5 was used. (TIF 3995 KB)
18_2023_4771_MOESM4_ESM.tif
Figure S4. Adipose tissue alterations among the groups. A. Browning-related genes (Pgc1a, Prdm16, Ucp1, Nrf1, and Cs) in the pgWAT. B. Browning-related genes (Pgc1a, Prdm16, Ucp1, Nrf1, Cs) in the ingWAT. C. Browning-related genes (Pgc1a, Prdm16, Ucp1, Nrf1, Cs) in the BAT. D. Representative images of hematoxylin–eosin (H&E, 10x) in the pgWAT. E. Representative images of hematoxylin–eosin (H&E, 10x) in the ingWAT. F. Representative images of hematoxylin–eosin (H&E, 10x, scale bar: 100 µM) in the BAT. In A-C, n = 5/group was used. # p < 0.05 vs Slc2a4+/− Sedentary group. (TIF 20533 KB)
18_2023_4771_MOESM5_ESM.tif
Figure S5. Indirect calorimetry among the groups. A. VO2 consumption during dark and light cycles, and Area under the curve (AUC). B. Heat production during dark and light cycle, and Area under the curve (AUC). C. VCO2 consumption during dark and light cycle, and Area under the curve (AUC). D. Accumulative food intake during dark and light cycle, and Area under the curve (AUC). E. Respiratory exchange ratio (RER) during dark and light cycle, and Area under the curve (AUC). F. Locomotor activity during dark and light cycle, and Area under the curve (AUC). In A-F, n = 5/group was used. * p < 0.05 vs WT group. # p < 0.05 vs Slc2a4+/− Sedentary group. (TIF 9250 KB)
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Muñoz, V.R., Botezelli, J.D., Gaspar, R.C. et al. Effects of short-term endurance and strength exercise in the molecular regulation of skeletal muscle in hyperinsulinemic and hyperglycemic Slc2a4+/− mice. Cell. Mol. Life Sci. 80, 122 (2023). https://doi.org/10.1007/s00018-023-04771-2
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DOI: https://doi.org/10.1007/s00018-023-04771-2