Diabetes Metab J.  2021 Sep;45(5):773-786. 10.4093/dmj.2020.0157.

The Effects of Exercise and Restriction of Sugar-Sweetened Beverages on Muscle Function and Autophagy Regulation in High-Fat High-Sucrose-Fed Obesity Mice

• Zhang D
• Lee JH
• Shin HE
• Kwak SE
• Bae JH
• Tang L
• Song W

• KMID: 2520859
• DOI: http://doi.org/10.4093/dmj.2020.0157

Abstract

Background
Autophagy maintains muscle mass and healthy skeletal muscles. Several recent studies have associated sugar-sweetened beverage (SSB) consumption with diseases. We investigated whether muscle dysfunction due to obesity could be restored by SSB restriction (SR) alone or in combination with exercise (EX) training.
Methods
Obese mice were subjected to SR combined with treadmill EX. Intraperitoneal glucose tolerance test, grip strength test, hanging time test, and body composition analysis were performed. Triglyceride (TG) and total cholesterol (TC) serum concentrations and TG concentrations in quadriceps muscles were analyzed. Western blot and reverse transcription-quantitative polymerase chain reaction helped analyze autophagy-related protein and mRNA expression, respectively.
Results
SR alone had no significant effect on fasting blood glucose levels, glucose tolerance, and muscle function. However, it had effect on serum TC, serum TG, and BCL2 interacting protein 3 expression. SR+EX improved glucose tolerance and muscle function and increased serum TC utilization than SR alone. SR+EX reduced P62 levels, increased glucose transporter type 4 and peroxisome proliferator-activated receptor γ coactivator-1α protein expression, and improved grip strength relative to the high-fat and high-sucrose liquid (HFHS) group, and this was not observed in the HFHS+EX group.
Conclusion
SR induced mitophagy-related protein expression in quadriceps, without affecting muscle function. And, the combination of SR and EX activated mitophagy-related proteins and improved muscle function.

Keyword

Autophagy; Diet, high-fat; Muscle, skeletal; Obesity; Sucrose; Sugar-sweetened beverages

Figure

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  Fig. 1. Schematic diagram of the experimental design. Male C57BL/6 mice, 4-week-old (n=61) were randomly assigned to six groups. Five mice from the high-fat and high-sucrose liquid (HFHS) group were subjected to glucose tolerance test and muscle performance test before re-grouping. After diet or exercise intervention, five mice from each of the eight groups were subjected to glucose tolerance test, muscle performance test, morphological observation, and tissue lipid quantification. Moreover, the other six mice from the groups as mentioned above were subjected to biological detection. Control (CON), normal diet+plain water; EX, exercise; SR, sugar-sweetened beverages (SSB) restriction, high-fat diet+plain water; TG, triglyceride; TC, total cholesterol; RT-PCR, reverse transcription-polymerase chain reaction.

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  Fig. 2. Effects of SR and exercise on body weight and food intake. (A) Images of mice. (B) Change in the body weight (n=5). (C) The area under the curve of energy intake (n=5). (D) Diet or liquid intake (n=5). (E) Wet weight of adipose tissues (n=6). (F) Normalized wet weight of adipose tissues (n=6). (G) Results of body composition analysis (n=5). All values are shown as the mean±standard deviation. HFHS, high-fat and high-sucrose liquid; EX, exercise; SR, sugar-sweetened beverages (SSB) restriction, high-fat diet+plain water; iWAT, inguinal white adipose tissue; eWAT, epididymal white adipose tissue; iBAT, interscapular brown adipose tissue. aP<0.05, bP<0.01 vs. the HFHS group and cP<0.05, dP<0.01 vs. the SR group.

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  Fig. 3. Effects of SR and exercise on glucose and lipid metabolism. (A) Fasting glucose (n=5). (B) Intraperitoneal glucose tolerance test (ipGTT) curve (n=5). (C) The area under the curve (AUC) of ipGTT (n=5). (D) Total cholesterol (TC) (n=6). (E) Triglyceride (TG) concentration (n=6) of serum. (F) TG concentration (n=6) of quadriceps (QUAD). All values are shown as the mean±standard deviation. 0 week, point before intervention; HFHS, high-fat and high-sucrose liquid; EX, exercise; SR, sugar-sweetened beverages (SSB) restriction, high-fat+plain water. aP<0.05, bP<0.01 vs. the HFHS group, cP<0.01 vs. the SR group, dP<0.05, eP<0.01 vs. 0 week, fP<0.01, and gP<0.01 vs. the control (CON) group

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  Fig. 4. Effects of SR and exercise on skeletal muscle performance. (A) Normalized quadriceps mass (n=6). (B) Normalized gastrocnemius mass (n=6). (C) Normalized tibialis anterior mass (n=6). (D) Normalized soleus mass (n=6). (E) Normalized extensor digitorum longus mass (n=6). (F) Normalized hanging time (n=5). (G) Normalized grip strength (n=5). All values are shown as the mean±standard deviation. 0 week, point before intervention; control (CON), normal diet+plain water; EX, exercise; HFHS, high-fat and high-sucrose liquid; SR, sugar-sweetened beverages (SSB) restriction, high-fat+plain water diet. aP<0.05, bP<0.01 vs. the HFHS group, cP<0.05, dP<0.01 vs. the SR group, eP<0.01 vs. 0 week, and fP<0.05, gP<0.01 vs. the CON group.

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  Fig. 5. Effects of SR and exercise on the expression of the autophagy pathway-related proteins. (A) Images of immunoblotting results. (B) The graph shows the expression of microtubule-associated proteins 1A/1B light chain 3 (LC3I), LC3II, and LC3II/LC3I ratio, (C) P62, (D) BCL2 interacting protein 3 (BNIP3), (E) beclin1, (F) glucose transporter type 4 (GLUT4), (G) myogenic differentiation (MyoD); (H) p-forkhead box O1 (FoxO1), (I) p-mechanistic target of rapamycin (mTOR)/mTOR ratio, (J) p-AMP-activated protein kinase (AMPK)/AMPK ratio, (K) peroxisome proliferator-activated receptor γ coactivator-1α (PGC1α), determined by densitometric quantification. All values are shown as the mean±standard error of the mean (n=6). GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Control (CON), normal diet+plain water; EX, exercise; HFHS, high-fat and high-sucrose liquid; SR, sugar-sweetened beverages (SSB) restriction, high-fat+plain water diet. aP<0.05, bP<0.01 vs. the HFHS group, cP<0.05, and dP<0.01 vs. the CON group.

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