Korean J Physiol Pharmacol.  2017 Nov;21(6):633-641. 10.4196/kjpp.2017.21.6.633.

Effects of intermittent ladder-climbing exercise training on mitochondrial biogenesis and endoplasmic reticulum stress of the cardiac muscle in obese middle-aged rats

• Kim K
• Ahn N
• Jung S
• Park S

• KMID: 2395258
• DOI: http://doi.org/10.4196/kjpp.2017.21.6.633

Abstract

The aim of this study is to investigate the effects of intermittent ladder-climbing exercise training on mitochondrial biogenesis and ER stress of the cardiac muscle in high fat diet-induced obese middle-aged rats. We induced obesity over 6 weeks of period in 40 male Sprague-Dawley rats around 50 weeks old, and were randomly divided into four experimental groups: chow, HFD, exercise+HFD, and exercise+chow. The exercising groups underwent high-intensity intermittent training using a ladder-climbing and weight exercise 3 days/week for a total of 8 weeks. High-fat diet and concurrent exercise resulted in no significant reduction in body weight but caused a significant reduction in visceral fat weight (p<0.05). Expression of PPARδ increased in the exercise groups and was significantly increased in the high-fat diet+exercise group (p<0.05). Among the ER stress-related proteins, the expression levels of p-PERK and CHOP, related to cardiac muscle damage, were significantly higher in the cardiac muscle of the high-fat diet group (p<0.05), and were significantly reduced by intermittent ladder-climbing exercise training (p<0.05). Specifically, this reduction was greater when the rats underwent exercise after switching back to the chow diet with a reduced caloric intake. Collectively, these results suggest that the combination of intermittent ladder-climbing exercise training and a reduced caloric intake can decrease the levels of ER stress-related proteins that contribute to cardiac muscle damage in obesity and aging. However, additional validation is required to understand the effects of these changes on mitochondrial biogenesis during exercise.

Keyword

Aging; Cardiac muscle; ER stress; High-fat diet-induced obesity; Ladder climbing exercise; Mitochondrial biogenesis

MeSH Terms

Aging
Animals
Body Weight
Diet
Diet, High-Fat
Endoplasmic Reticulum Stress*
Endoplasmic Reticulum*
Energy Intake
Humans
Intra-Abdominal Fat
Male
Myocardium*
Obesity
Organelle Biogenesis*
Rats*
Rats, Sprague-Dawley

Figure

• 

  Fig. 1 Body weight and visceral fat weight. (A) Changes in body weight after 8-week intervention. αSignificantly different from CD, CE (p<0.05). (B) Changes of visceral fat weight after 8-week intervention. +significantly different from CD, CE (p<0.05); *Significantly different from CD, CE, FE (p<0.05). CD, normal chow diet; CE, chow diet+exercise; FD, high-fat diet only; FE, high-fat diet+exercise.

• 

  Fig. 2 Expression levels of mitochondrial enzymes in the cardiac muscles. +Significantly different from CD, FD (p<0.05); *Significantly different from CD, FD, FE (p<0.05). CD, normal chow diet; CE, chow diet+exercise; FD, high-fat diet only; FE, high-fat diet+exercise. SUD, succinate dehydrogenase (A); Cyto C (B), cytochrome C.

• 

  Fig. 3 Changes in PPARβ (A) and PGC-1α (B) protein expression in the cardiac muscle after 8-week diet and/or exercise intervention. *Significantly different from CD, CE, FD (p<0.05). CD, normal chow diet; CE, chow diet+exercise; FD, high-fat diet only; FE, high-fat diet+exercise.

• 

  Fig. 4 Changes in (A) p-PERK, and (B) GRP78 (blue) and CHOP (red) protein expression levels in the cardiac muscle after 8 weeks of diet and/or exercise intervention. (A) +Significantly different from CD (p < 0.05); *Significantly different from CD, CE, FE (p<0.05). (B) +Significantly different from CD, FE (p<0.05); *Significantly different from CD, CE, FE (p<0.05). CD, normal chow diet; CE, chow diet+exercise; FD, high-fat diet only; FE, high-fat diet+exercise.

• 

  Fig. 5 Effect of ladder-climbing exercise training on mitochondrial biogenesis and ER stress of the cardiac muscle. We put forward a hypothesis that the concurrent stimulation from exercise training is more effective through inhibition of PERK and CHOP expression or the activation of PGC-1α and PPARb in protecting the cardiac muscle compared to a weight loss scheme that is entirely dependent on diet and reduced caloric intake. However, we could not suggest the significant relationship between PGC-1α and PPARβ and the activation GRP78 expression.

Reference

1. Higami Y, Shimokawa I. Apoptosis in the aging process. Cell Tissue Res. 2000; 301:125–132.

2. Di Lisa F, Bernardi P. Mitochondrial function and myocardial aging. A critical analysis of the role of permeability transition. Cardiovasc Res. 2005; 66:222–232.

3. Rimbaud S, Garnier A, Ventura-Clapier R. Mitochondrial biogenesis in cardiac pathophysiology. Pharmacol Rep. 2009; 61:131–138.

4. Palomer X, Capdevila-Busquets E, Botteri G, Salvadó L, Barroso E, Davidson MM, Michalik L, Wahli W, Vázquez-Carrera M. PPARβ/δ attenuates palmitate-induced endoplasmic reticulum stress and induces autophagic markers in human cardiac cells. Int J Cardiol. 2014; 174:110–118.

5. Colgan SM, Tang D, Werstuck GH, Austin RC. Endoplasmic reticulum stress causes the activation of sterol regulatory element binding protein-2. Int J Biochem Cell Biol. 2007; 39:1843–1851.

6. Myoishi M, Hao H, Minamino T, Watanabe K, Nishihira K, Hatakeyama K, Asada Y, Okada K, Ishibashi-Ueda H, Gabbiani G, Bochaton-Piallat ML, Mochizuki N, Kitakaze M. Increased endoplasmic reticulum stress in atherosclerotic plaques associated with acute coronary syndrome. Circulation. 2007; 116:1226–1233.

7. Wu J, Kaufman RJ. From acute ER stress to physiological roles of the Unfolded Protein Response. Cell Death Differ. 2006; 13:374–384.

8. Nisha VM, Anusree SS, Priyanka A, Raghu KG. Apigenin and quercetin ameliorate mitochondrial alterations by tunicamycin-induced ER stress in 3T3-L1 adipocytes. Appl Biochem Biotechnol. 2014; 174:1365–1375.

9. Verfaillie T, Rubio N, Garg AD, Bultynck G, Rizzuto R, Decuypere JP, Piette J, Linehan C, Gupta S, Samali A, Agostinis P. PERK is required at the ER-mitochondrial contact sites to convey apoptosis after ROS-based ER stress. Cell Death Differ. 2012; 19:1880–1891.

10. Holloszy JO. Regulation by exercise of skeletal muscle content of mitochondria and GLUT4. J Physiol Pharmacol. 2008; 59:Suppl 7. 5–18.

11. Terada S, Yokozeki T, Kawanaka K, Ogawa K, Higuchi M, Ezaki O, Tabata I. Effects of high-intensity swimming training on GLUT-4 and glucose transport activity in rat skeletal muscle. J Appl Physiol (1985). 2001; 90:2019–2024.

12. Coggan AR, Spina RJ, King DS, Rogers MA, Brown M, Nemeth PM, Holloszy JO. Skeletal muscle adaptations to endurance training in 60- to 70-yr-old men and women. J Appl Physiol (1985). 1992; 72:1780–1786.

13. Kohrt WM, Malley MT, Coggan AR, Spina RJ, Ogawa T, Ehsani AA, Bourey RE, Martin WH 3rd, Holloszy JO. Effects of gender, age, and fitness level on response of VO2max to training in 60-71 yr olds. J Appl Physiol (1985). 1991; 71:2004–2011.

14. Short KR. Changes of deleted mtDNA after endurance exercise. Eur J Appl Physiol. 2003; 90:223.

15. Jubrias SA, Esselman PC, Price LB, Cress ME, Conley KE. Large energetic adaptations of elderly muscle to resistance and endurance training. J Appl Physiol (1985). 2001; 90:1663–1670.

16. Balagopal P, Schimke JC, Ades P, Adey D, Nair KS. Age effect on transcript levels and synthesis rate of muscle MHC and response to resistance exercise. Am J Physiol Endocrinol Metab. 2001; 280:E203–E208.

17. Hasten DL, Pak-Loduca J, Obert KA, Yarasheski KE. Resistance exercise acutely increases MHC and mixed muscle protein synthesis rates in 78-84 and 23-32 yr olds. Am J Physiol Endocrinol Metab. 2000; 278:E620–E626.

18. Menshikova EV, Ritov VB, Fairfull L, Ferrell RE, Kelley DE, Goodpaster BH. Effects of exercise on mitochondrial content and function in aging human skeletal muscle. J Gerontol A Biol Sci Med Sci. 2006; 61:534–540.

19. Kavazis AN, McClung JM, Hood DA, Powers SK. Exercise induces a cardiac mitochondrial phenotype that resists apoptotic stimuli. Am J Physiol Heart Circ Physiol. 2008; 294:H928–H935.

20. Hamilton KL, Powers SK, Sugiura T, Kim S, Lennon S, Tumer N, Mehta JL. Short-term exercise training can improve myocardial tolerance to I/R without elevation in heat shock proteins. Am J Physiol Heart Circ Physiol. 2001; 281:H1346–H1352.

21. Morán M, Delgado J, González B, Manso R, Megías A. Responses of rat myocardial antioxidant defences and heat shock protein HSP72 induced by 12 and 24-week treadmill training. Acta Physiol Scand. 2004; 180:157–166.

22. Radak Z, Taylor AW, Ohno H, Goto S. Adaptation to exercise-induced oxidative stress: from muscle to brain. Exerc Immunol Rev. 2001; 7:90–107.

23. Starnes JW, Taylor RP. Exercise-induced cardioprotection: endogenous mechanisms. Med Sci Sports Exerc. 2007; 39:1537–1543.

24. Ding G, Fu M, Qin Q, Lewis W, Kim HW, Fukai T, Bacanamwo M, Chen YE, Schneider MD, Mangelsdorf DJ, Evans RM, Yang Q. Cardiac peroxisome proliferator-activated receptor γ is essential in protecting cardiomyocytes from oxidative damage. Cardiovasc Res. 2007; 76:269–279.

25. Girnun GD, Domann FE, Moore SA, Robbins ME. Identification of a functional peroxisome proliferator-activated receptor response element in the rat catalase promoter. Mol Endocrinol. 2002; 16:2793–2801.

26. Fan W, Waizenegger W, Lin CS, Sorrentino V, He MX, Wall CE, Li H, Liddle C, Yu RT, Atkins AR, Auwerx J, Downes M, Evans RM. PPARδ promotes running endurance by preserving glucose. Cell Metab. 2017; 25:1186–1193.e4.

27. Cheang WS, Tian XY, Wong WT, Lau CW, Lee SS, Chen ZY, Yao X, Wang N, Huang Y. Metformin protects endothelial function in diet-induced obese mice by inhibition of endoplasmic reticulum stress through 5' adenosine monophosphate-activated protein kinase-peroxisome proliferator-activated receptor δ pathway. Arterioscler Thromb Vasc Biol. 2014; 34:830–836.

28. Cheang WS, Wong WT, Zhao L, Xu J, Wang L, Lau CW, Chen ZY, Ma RC, Xu A, Wang N, Tian XY, Huang Y. PPARδ is required for exercise to attenuate endoplasmic reticulum stress and endothelial dysfunction in diabetic mice. Diabetes. 2017; 66:519–528.

29. Hancock CR, Han DH, Chen M, Terada S, Yasuda T, Wright DC, Holloszy JO. High-fat diets cause insulin resistance despite an increase in muscle mitochondria. Proc Natl Acad Sci U S A. 2008; 105:7815–7820.

30. Kraemer WJ, Flanagan SD, Volek JS, Nindl BC, Vingren JL, Dunn-Lewis C, Comstock BA, Hooper DR, Szivak TK, Looney DP, Maresh CM, Hymer WC. Resistance exercise induces region-specific adaptations in anterior pituitary gland structure and function in rats. J Appl Physiol (1985). 2013; 115:1641–1647.

31. Lee S, Barton ER, Sweeney HL, Farrar RP. Viral expression of insulin-like growth factor-I enhances muscle hypertrophy in resistance-trained rats. J Appl Physiol (1985). 2004; 96:1097–1104.

32. Jung S, Ahn N, Kim S, Byun J, Joo Y, Kim S, Jung Y, Park S, Hwang I, Kim K. The effect of ladder-climbing exercise on atrophy/hypertrophy-related myokine expression in middle-aged male Wistar rats. J Physiol Sci. 2015; 65:515–521.

33. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding. Anal Biochem. 1976; 72:248–254.

34. Ozcan U, Cao Q, Yilmaz E, Lee AH, Iwakoshi NN, Ozdelen E, Tuncman G, Görgün C, Glimcher LH, Hotamisligil GS. Endoplasmic reticulum stress links obesity, insulin action, and type 2 diabetes. Science. 2004; 306:457–461.

35. Mori K. Tripartite management of unfolded proteins in the endoplasmic reticulum. Cell. 2000; 101:451–454.

36. Oyadomari S, Koizumi A, Takeda K, Gotoh T, Akira S, Araki E, Mori M. Targeted disruption of the Chop gene delays endoplasmic reticulum stress-mediated diabetes. J Clin Invest. 2002; 109:525–532.

37. Schröder M, Kaufman RJ. The mammalian unfolded protein response. Annu Rev Biochem. 2005; 74:739–789.

38. Murlasits Z, Lee Y, Powers SK. Short-term exercise does not increase ER stress protein expression in cardiac muscle. Med Sci Sports Exerc. 2007; 39:1522–1528.

39. González B, Hernando R, Manso R. Stress proteins of 70 kDa in chronically exercised skeletal muscle. Pflugers Arch. 2000; 440:42–49.

40. Vitadello M, Penzo D, Petronilli V, Michieli G, Gomirato S, Menabò R, Di Lisa F, Gorza L. Overexpression of the stress protein Grp94 reduces cardiomyocyte necrosis due to calcium overload and simulated ischemia. FASEB J. 2003; 17:923–925.

41. Zhang J, Jiang Y, Jia Z, Li Q, Gong W, Wang L, Wei D, Yao J, Fang S, Xie K. Association of elevated GRP78 expression with increased lymph node metastasis and poor prognosis in patients with gastric cancer. Clin Exp Metastasis. 2006; 23:401–410.

42. Baar K, Wende AR, Jones TE, Marison M, Nolte LA, Chen M, Kelly DP, Holloszy JO. Adaptations of skeletal muscle to exercise: rapid increase in the transcriptional coactivator PGC-1. FASEB J. 2002; 16:1879–1886.

43. Goto M, Terada S, Kato M, Katoh M, Yokozeki T, Tabata I, Shimokawa T. cDNA Cloning and mRNA analysis of PGC-1 in epitrochlearis muscle in swimming-exercised rats. Biochem Biophys Res Commun. 2000; 274:350–354.

44. Higashida T, Kreipke CW, Rafols JA, Peng C, Schafer S, Schafer P, Ding JY, Dornbos D 3rd, Li X, Guthikonda M, Rossi NF, Ding Y. The role of hypoxia-inducible factor-1α, aquaporin-4, and matrix metalloproteinase-9 in blood-brain barrier disruption and brain edema after traumatic brain injury. J Neurosurg. 2011; 114:92–101.

45. Terada S, Goto M, Kato M, Kawanaka K, Shimokawa T, Tabata I. Effects of low-intensity prolonged exercise on PGC-1 mRNA expression in rat epitrochlearis muscle. Biochem Biophys Res Commun. 2002; 296:350–354.

46. Terada S, Tabata I. Effects of acute bouts of running and swimming exercise on PGC-1alpha protein expression in rat epitrochlearis and soleus muscle. Am J Physiol Endocrinol Metab. 2004; 286:E208–E216.

47. Wende AR, Huss JM, Schaeffer PJ, Giguère V, Kelly DP. PGC-1alpha coactivates PDK4 gene expression via the orphan nuclear receptor ERRalpha: a mechanism for transcriptional control of muscle glucose metabolism. Mol Cell Biol. 2005; 25:10684–10694.

48. Wang YX, Lee CH, Tiep S, Yu RT, Ham J, Kang H, Evans RM. Peroxisome-proliferator-activated receptor delta activates fat metabolism to prevent obesity. Cell. 2003; 113:159–170.

49. Braissant O, Foufelle F, Scotto C, Dauça M, Wahli W. Differential expression of peroxisome proliferator-activated receptors (PPARs): tissue distribution of PPAR-alpha, -beta, and -gamma in the adult rat. Endocrinology. 1996; 137:354–366.

50. Luquet S, Lopez-Soriano J, Holst D, Fredenrich A, Melki J, Rassoulzadegan M, Grimaldi PA. Peroxisome proliferator-activated receptor delta controls muscle development and oxidative capability. FASEB J. 2003; 17:2299–2301.

51. Tanaka T, Yamamoto J, Iwasaki S, Asaba H, Hamura H, Ikeda Y, Watanabe M, Magoori K, Ioka RX, Tachibana K, Watanabe Y, Uchiyama Y, Sumi K, Iguchi H, Ito S, Doi T, Hamakubo T, Naito M, Auwerx J, Yanagisawa M, Kodama T, Sakai J. Activation of peroxisome proliferator-activated receptor delta induces fatty acid beta-oxidation in skeletal muscle and attenuates metabolic syndrome. Proc Natl Acad Sci U S A. 2003; 100:15924–15929.

52. Wang YX, Zhang CL, Yu RT, Cho HK, Nelson MC, Bayuga-Ocampo CR, Ham J, Kang H, Evans RM. Regulation of muscle fiber type and running endurance by PPARdelta. PLoS Biol. 2004; 2:e294.

53. Kelly DP, Scarpulla RC. Transcriptional regulatory circuits controlling mitochondrial biogenesis and function. Genes Dev. 2004; 18:357–368.

54. Scarpulla RC. Nuclear activators and coactivators in mammalian mitochondrial biogenesis. Biochim Biophys Acta. 2002; 1576:1–14.

55. Koh JH, Hancock CR, Terada S, Higashida K, Holloszy JO, Han DH. PPARβ is essential for maintaining normal levels of PGC-1α and mitochondria and for the increase in muscle mitochondria induced by exercise. Cell Metab. 2017; 25:1176–1185.e5.

56. Anderson EJ, Lustig ME, Boyle KE, Woodlief TL, Kane DA, Lin CT, Price JW 3rd, Kang L, Rabinovitch PS, Szeto HH, Houmard JA, Cortright RN, Wasserman DH, Neufer PD. Mitochondrial H2O2 emission and cellular redox state link excess fat intake to insulin resistance in both rodents and humans. J Clin Invest. 2009; 119:573–581.

57. Barbosa MR, Sampaio IH, Teodoro BG, Sousa TA, Zoppi CC, Queiroz AL, Passos MA, Alberici LC, Teixeira FR, Manfiolli AO, Batista TM, Cappelli AP, Reis RI, Frasson D, Kettelhut IC, Parreirase-Silva LT, Costa-Neto CM, Carneiro EM, Curi R, Silveira LR. Hydrogen peroxide production regulates the mitochondrial function in insulin resistant muscle cells: effect of catalase overexpression. Biochim Biophys Acta. 2013; 1832:1591–1604.

58. Ding H, Heng B, He W, Shi L, Lai C, Xiao L, Ren H, Mo S, Su Z. Chronic reactive oxygen species exposure inhibits glucose uptake and causes insulin resistance in C2C12 myotubes. Biochem Biophys Res Commun. 2016; 478:798–803.

59. Henriksen EJ. Effects of H2O2 on insulin signaling the glucose transport system in mammalian skeletal muscle. Methods Enzymol. 2013; 528:269–278.

60. Malhotra JD, Kaufman RJ. Endoplasmic reticulum stress and oxidative stress: a vicious cycle or a double-edged sword? Antioxid Redox Signal. 2007; 9:2277–2293.

61. Pinton P, Giorgi C, Pandolfi PP. The role of PML in the control of apoptotic cell fate: a new key player at ER-mitochondria sites. Cell Death Differ. 2011; 18:1450–1456.

62. Sano R, Annunziata I, Patterson A, Moshiach S, Gomero E, Opferman J, Forte M, d'Azzo A. GM1-ganglioside accumulation at the mitochondria-associated ER membranes links ER stress to Ca²⁺-dependent mitochondrial apoptosis. Mol Cell. 2009; 36:500–511.

63. Hayashi T, Rizzuto R, Hajnoczky G, Su TP. MAM: more than just a housekeeper. Trends Cell Biol. 2009; 19:81–88.

64. Hunter RB, Mitchell-Felton H, Essig DA, Kandarian SC. Expression of endoplasmic reticulum stress proteins during skeletal muscle disuse atrophy. Am J Physiol Cell Physiol. 2001; 281:C1285–C1290.

65. Chen Y, Pandiri I, Joe Y, Kim HJ, Kim SK, Park J, Ryu J, Cho GJ, Park JW, Ryter SW, Chung HT. Synergistic effects of cilostazol and probucol on ER stress-induced hepatic steatosis via heme oxygenase-1-dependent activation of mitochondrial biogenesis. Oxid Med Cell Longev. 2016; 2016:3949813.

66. St-Pierre J, Drori S, Uldry M, Silvaggi JM, Rhee J, Jäger S, Handschin C, Zheng K, Lin J, Yang W, Simon DK, Bachoo R, Spiegelman BM. Suppression of reactive oxygen species and neurodegeneration by the PGC-1 transcriptional coactivators. Cell. 2006; 127:397–408.

67. Civitarese AE, Carling S, Heilbronn LK, Hulver MH, Ukropcova B, Deutsch WA, Smith SR, Ravussin E. Calorie restriction increases muscle mitochondrial biogenesis in healthy humans. PLoS Med. 2007; 4:e76.

68. Ballor DL. Effect of dietary restriction and/or exercise on 23-h metabolic rate and body composition in female rats. J Appl Physiol (1985). 1991; 71:801–806.

69. Dulloo AG, Girardier L. 24 hour energy expenditure several months after weight loss in the underfed rat: evidence for a chronic increase in whole-body metabolic efficiency. Int J Obes Relat Metab Disord. 1993; 17:115–123.

70. McCarter RJ, Palmer J. Energy metabolism and aging: a lifelong study of Fischer 344 rats. Am J Physiol. 1992; 263:E448–E452.

71. Nisoli E, Tonello C, Cardile A, Cozzi V, Bracale R, Tedesco L, Falcone S, Valerio A, Cantoni O, Clementi E, Moncada S, Carruba MO. Calorie restriction promotes mitochondrial biogenesis by inducing the expression of eNOS. Science. 2005; 310:314–317.

72. Kempen GI, Miedema I, van den Bos GA, Ormel J. Relationship of domain-specific measures of health to perceived overall health among older subjects. J Clin Epidemiol. 1998; 51:11–18.

73. Simoneau JA, Veerkamp JH, Turcotte LP, Kelley DE. Markers of capacity to utilize fatty acids in human skeletal muscle: relation to insulin resistance and obesity and effects of weight loss. FASEB J. 1999; 13:2051–2060.