Changes in UPR-PERK pathway and muscle hypertrophy following resistance training and creatine supplementation in rats

Abstract

The unfolded protein response (UPR) plays a pivotal role in some exercise training–induced physiological adaptation. Our aim was to evaluate the changes in the protein kinase R-like endoplasmic reticulum kinase (PERK) arm of the UPR and hypertrophy signaling pathway following 8 weeks of resistance training and creatine (Cr) supplementation in rats. Thirty-two adult male Wistar rats (8 weeks old) were randomly divided into 4 groups of 8: untrained + placebo (UN+P), resistance training + placebo (RT+P), untrained + Cr (UN+Cr), and resistance training + Cr (RT+Cr). Trained animals were submitted to the ladder-climbing exercise training 5 days per week for a total of 8 weeks. Cr supplementation groups received creatine diluted with 1.5 ml of 5% dextrose orally. The flexor hallucis longus (FHL) muscle was extracted 48 h after the last training session and used for western blotting. After training period, the RT+Cr and RT+P groups presented a significant increase in phosphorylated and phosphorylated/total ratio hypertrophy indices, phosphorylated and phosphorylated/total ratio PERK pathway proteins, and other downstream proteins of the PERK cascade compared with their untrained counterparts (P < 0.05). The increase in hypertrophy indices were higher but PERK pathway proteins were lower in the RT-Cr group than in the RT+P group (P < 0.05). There was no significant difference between the untrained groups (P > 0.05). Our study suggests that resistance training in addition to Cr supplementation modifies PERK pathway response and improves skeletal muscle hypertrophy.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

References

  1. 1.

    Aguiar AF, de Souza RWA, Aguiar DH, Aguiar RCM, Vechetti IJ Jr, Dal-Pai-Silva M (2011) Creatine does not promote hypertrophy in skeletal muscle in supplemented compared with nonsupplemented rats subjected to a similar workload. Nutr Res 31:652–657. https://doi.org/10.1016/j.nutres.2011.08.006

    CAS  Article  PubMed  Google Scholar 

  2. 2.

    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1016/0003-2697(76)90527-3

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  3. 3.

    Buford TW, Kreider RB, Stout JR, Greenwood M, Campbell B, Spano M, Ziegenfuss T, Lopez H, Landis J, Antonio J (2007) International Society of Sports Nutrition position stand: creatine supplementation and exercise. J Int Soc Sports Nutr 4:6. https://doi.org/10.1186/1550-2783-4-6

    Article  PubMed  PubMed Central  Google Scholar 

  4. 4.

    Burke DG, Candow DG, Chilibeck PD, MacNeil LG, Roy BD, Tarnopolsky MA, Ziegenfuss T (2008) Effect of creatine supplementation and resistance-exercise training on muscle insulin-like growth factor in young adults. Int J Sport Nutr Exerc Metab 18:389–398. https://doi.org/10.1123/ijsnem.18.4.389

    CAS  Article  PubMed  Google Scholar 

  5. 5.

    Cunha MP, Budni J, Ludka FK, Pazini FL, Rosa JM, Oliveira Á, Lopes MW, Tasca CI, Leal RB, Rodrigues ALS (2016) Involvement of PI3K/Akt signaling pathway and its downstream intracellular targets in the antidepressant-like effect of creatine. Mol Neurobiol 53:2954–2968. https://doi.org/10.1007/s12035-015-9192-4

    CAS  Article  PubMed  Google Scholar 

  6. 6.

    Deldicque L, Cani PD, Delzenne NM, Baar K, Francaux M (2013) Endurance training in mice increases the unfolded protein response induced by a high-fat diet. J Physiol Biochem 69:215–225. https://doi.org/10.1007/s13105-012-0204-9

    CAS  Article  PubMed  Google Scholar 

  7. 7.

    Deminice R, Jordao AA (2012) Creatine supplementation reduces oxidative stress biomarkers after acute exercise in rats. Amino Acids 43:709–715. https://doi.org/10.1007/s00726-011-1121-x

    CAS  Article  PubMed  Google Scholar 

  8. 8.

    Drummond MJ, Fry CS, Glynn EL, Timmerman KL, Dickinson JM, Walker DK, Gundermann DM, Volpi E, Rasmussen BB (2011) Skeletal muscle amino acid transporter expression is increased in young and older adults following resistance exercise. J Appl Physiol 111:135–142. https://doi.org/10.1152/japplphysiol.01408.2010

    CAS  Article  PubMed  Google Scholar 

  9. 9.

    Estébanez B, Moreira OC, Almar M, de Paz JA, Gonzalez-Gallego J, Cuevas MJ (2019) Effects of a resistance-training programme on endoplasmic reticulum unfolded protein response and mitochondrial functions in PBMCs from elderly subjects. Eur J Sport Sci 19:931–940. https://doi.org/10.1080/17461391.2018.1561950

    Article  PubMed  Google Scholar 

  10. 10.

    Ferretti R, Moura EG, Dos Santos VC, Caldeira EJ, Conte M, Matsumura CY, Pertille A, Mosqueira M (2018) High-fat diet suppresses the positive effect of creatine supplementation on skeletal muscle function by reducing protein expression of IGF-PI3K-AKT-mTOR pathway. PLoS One 13:e0199728. https://doi.org/10.1371/journal.pone.0199728

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  11. 11.

    Fujii J, Homma T, Kobayashi S, Seo HG (2018) Mutual interaction between oxidative stress and endoplasmic reticulum stress in the pathogenesis of diseases specifically focusing on non-alcoholic fatty liver disease. World J Biol Chem 9:1–15. https://doi.org/10.4331/wjbc.v9.i1.1

    Article  PubMed  PubMed Central  Google Scholar 

  12. 12.

    Gallot YS, Bohnert KR, Straughn AR, Xiong G, Hindi SM, Kumar A (2018) PERK regulates skeletal muscle mass and contractile function in adult mice. FASEB J 33:1946–1962. https://doi.org/10.1096/fj.201800683RR

    Article  PubMed  PubMed Central  Google Scholar 

  13. 13.

    Hentilä J, Ahtiainen J, Paulsen G, Raastad T, Häkkinen K, Mero A, Hulmi J (2018) Autophagy is induced by resistance exercise in young men, but unfolded protein response is induced regardless of age. Acta Physiol 224:e13069. https://doi.org/10.1111/apha.13069

    CAS  Article  Google Scholar 

  14. 14.

    Hespel P, Derave W (2007) Ergogenic effects of creatine in sports and rehabilitation. In: Creatine and creatine kinase in health and disease. Springer, pp 246–259. https://doi.org/10.1007/978-1-4020-6486-9_12

  15. 15.

    Hickner RC, Dyck DJ, Sklar J, Hatley H, Byrd P (2010) Effect of 28 days of creatine ingestion on muscle metabolism and performance of a simulated cycling road race. J Int Soc Sports Nutr 7:26. https://doi.org/10.1186/1550-2783-7-26

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  16. 16.

    Hornberger TA Jr, Farrar RP (2004) Physiological hypertrophy of the FHL muscle following 8 weeks of progressive resistance exercise in the rat. Can J Appl Physiol 29:16–31. https://doi.org/10.1139/h04-002

    CAS  Article  PubMed  Google Scholar 

  17. 17.

    Hotamisligil GS (2010) Endoplasmic reticulum stress and the inflammatory basis of metabolic disease. Cell 140:900–917. https://doi.org/10.1016/j.cell.2010.02.034

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  18. 18.

    Kim K, Kim Y-H, Lee S-H, Jeon M-J, Park S-Y, Doh K-O (2014) Effect of exercise intensity on unfolded protein response in skeletal muscle of rat. Korean J Physiol Pharmacol 18:211–216. https://doi.org/10.4196/kjpp.2014.18.3.211

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  19. 19.

    Kim K, Ahn N, Jung S (2018) Comparison of endoplasmic reticulum stress and mitochondrial biogenesis responses after 12 weeks of treadmill running and ladder climbing exercises in the cardiac muscle of middle-aged obese rats. Braz J Med Biol Res 51. https://doi.org/10.1590/1414-431x20187508

  20. 20.

    Kwon I, Jang Y, Cho J-Y, Jang YC, Lee Y (2018) Long-term resistance exercise-induced muscular hypertrophy is associated with autophagy modulation in rats. J Physiol Sci 68:269–280. https://doi.org/10.1007/s12576-017-0531-2

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Liang S-H, Zhang W, Mcgrath BC, Zhang P, Cavener DR (2006) PERK (eIF2α kinase) is required to activate the stress-activated MAPKs and induce the expression of immediate-early genes upon disruption of ER calcium homoeostasis. Biochem J 393:201–209. https://doi.org/10.1042/BJ20050374

    CAS  Article  PubMed  Google Scholar 

  22. 22.

    Memme JM, Oliveira AN, Hood DA (2016) Chronology of UPR activation in skeletal muscle adaptations to chronic contractile activity. Am J Phys Cell Phys 310:C1024–C1036. https://doi.org/10.1152/ajpcell.00009.2016

    Article  Google Scholar 

  23. 23.

    Miyake M, Nomura A, Ogura A, Takehana K, Kitahara Y, Takahara K, Tsugawa K, Miyamoto C, Miura N, Sato R (2016) Skeletal muscle–specific eukaryotic translation initiation factor 2α phosphorylation controls amino acid metabolism and fibroblast growth factor 21–mediated non–cell-autonomous energy metabolism. FASEB J 30:798–812. https://doi.org/10.1096/fj.15-275990

    CAS  Article  PubMed  Google Scholar 

  24. 24.

    Novoa I, Zeng H, Harding HP, Ron D (2001) Feedback inhibition of the unfolded protein response by GADD34-mediated dephosphorylation of eIF2α. J Cell Biol 153:1011–1022. https://doi.org/10.1083/jcb.153.5.1011

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  25. 25.

    Ogasawara R, Fujita S, Hornberger TA, Kitaoka Y, Makanae Y, Nakazato K, Naokata I (2016) The role of mTOR signalling in the regulation of skeletal muscle mass in a rodent model of resistance exercise. Sci Rep 6:31142. https://doi.org/10.1038/srep31142

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  26. 26.

    Ogborn DI, McKay BR, Crane JD, Parise G, Tarnopolsky MA (2014) The unfolded protein response is triggered following a single, unaccustomed resistance-exercise bout. Am J Phys Regul Integr Comp Phys 307:R664–R669. https://doi.org/10.1152/ajpregu.00511.2013

    CAS  Article  Google Scholar 

  27. 27.

    Olsen S, Aagaard P, Kadi F, Tufekovic G, Verney J, Olesen JL, Suetta C, Kjær M (2006) Creatine supplementation augments the increase in satellite cell and myonuclei number in human skeletal muscle induced by strength training. J Physiol 573:525–534. https://doi.org/10.1113/jphysiol.2006.107359

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  28. 28.

    Pereira BC, Da Rocha AL, Pinto AP, Pauli JR, De Souza CT, Cintra DE, Ropelle ER, De Freitas EC, Zagatto AM, Da Silva AS (2016) Excessive eccentric exercise-induced overtraining model leads to endoplasmic reticulum stress in mice skeletal muscles. Life Sci 145:144–151. https://doi.org/10.1016/j.lfs.2015.12.037

    CAS  Article  PubMed  Google Scholar 

  29. 29.

    Sano R, Reed JC (2013) ER stress-induced cell death mechanisms. Biochim Biophys Acta (BBA)-Mol Cell Res 1833:3460–3470. https://doi.org/10.1016/j.bbamcr.2013.06.028

    CAS  Article  Google Scholar 

  30. 30.

    Schröder M, Kaufman RJ (2005) ER stress and the unfolded protein response. Mutat Res/Fundam Mol Mech Mutagen 569:29–63. https://doi.org/10.1016/j.mrfmmm.2004.06.056

    CAS  Article  Google Scholar 

  31. 31.

    Smiles WJ, Hawley JA, Camera DM (2016) Effects of skeletal muscle energy availability on protein turnover responses to exercise. J Exp Biol 219:214–225. https://doi.org/10.1242/jeb.125104

    Article  PubMed  Google Scholar 

  32. 32.

    Stefani GP, Nunes RB, Dornelles AZ, Alves JP, Piva MO, Di Domenico M, Rhoden CR, Dal Lago P (2014) Effects of creatine supplementation associated with resistance training on oxidative stress in different tissues of rats. J Int Soc Sports Nutr 11:11. https://doi.org/10.1186/1550-2783-11-11

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  33. 33.

    Takegaki J, Ogasawara R, Tamura Y, Takagi R, Arihara Y, Tsutaki A, Nakazato K, Ishii N (2017) Repeated bouts of resistance exercise with short recovery periods activates mTOR signaling, but not protein synthesis, in mouse skeletal muscle. Phys Rep 5:e13515. https://doi.org/10.14814/phy2.13515

    CAS  Article  Google Scholar 

  34. 34.

    Wu J, Kaufman R (2006) From acute ER stress to physiological roles of the unfolded protein response. Cell Death Differ 13:374–384. https://doi.org/10.1038/sj.cdd.4401840

    CAS  Article  PubMed  Google Scholar 

  35. 35.

    Wu J, Ruas JL, Estall JL, Rasbach KA, Choi JH, Ye L, Boström P, Tyra HM, Crawford RW, Campbell KP (2011) The unfolded protein response mediates adaptation to exercise in skeletal muscle through a PGC-1α/ATF6α complex. Cell Metab 13:160–169. https://doi.org/10.1016/j.cmet.2011.01.003

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  36. 36.

    Xiong G, Hindi SM, Mann AK, Gallot YS, Bohnert KR, Cavener DR, Whittemore SR, Kumar A (2017) The PERK arm of the unfolded protein response regulates satellite cell-mediated skeletal muscle regeneration. Elife 6:e22871. https://doi.org/10.7554/eLife.22871.001

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Funding

This research work was financially supported by the University of Kurdistan, Sanandaj, Iran (grant number: 17873).

Author information

Affiliations

Authors

Contributions

Hersh N and Dariush SH-V performed the experiments, analyzed the data, and drafted the manuscript. Dariush SH-V designed the research. Mohammad Raman M conducted the histological assays and participated in the writing of the manuscript. The authors declare that all data were generated in-house and that no paper mill was used.

Corresponding author

Correspondence to Dariush Sheikholeslami-Vatani.

Ethics declarations

Ethics approval

Animals used in the current research were treated in accordance with the National Institutes of Health Guide for the care and use of laboratory animals, and the study design was reviewed and approved by the Animal Research Ethics Committee of Kurdistan University of Medical Sciences (IR.MUK.REC.1397.5010).

Conflict of interest

The authors declare no competing interests.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Key Points

Creatine has a synergistic effect on hypertrophy caused by resistance training.

Resistance training increases phosphorylates PERK proteins and its downstream pathway.

Creatine supplementation during resistance training modifies PERK pathway response.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Nouri, H., Sheikholeslami-Vatani, D. & Moloudi, M.R. Changes in UPR-PERK pathway and muscle hypertrophy following resistance training and creatine supplementation in rats. J Physiol Biochem 77, 331–339 (2021). https://doi.org/10.1007/s13105-021-00801-4

Download citation

Keywords

  • Ladder-climbing training
  • UPR
  • PERK pathway
  • Muscle mass
  • Creatine