Journal of Sport and Exercise Physiology

Journal of Sport and Exercise Physiology

Effects of intermittent cycles of ischemia with resistance and endurance training on Murf-1 and Atrogin-1 gene expression and fiber diameter of gastrocnemius muscle in diabetic rats

Document Type : Original Article

Authors
Majid Zatparvar
Amin Farzaneh Hesari
Parvin Farzanegi
Department of Exercise Physiology, Sari Branch, Islamic Azad University, Sari, Iran
10.48308/joeppa.2024.235584.1250
Abstract
Background and Purpose: Muscular atrophy is one of the most common complications of diabetes. Exercise training has been suggested as one of the treatment strategies for muscular atrophy in diabetes. Intermittent cycles of ischemia (ICI) protects tissues from subsequent ischemic injury. However, whether ICI has beneficial effects on diabetic atrophy remains unknown. The present study aimed to investigate the synergistic effect of ICI with resistance and endurance training on Murf-1 and Atrogin-1 gene expression and fiber diameter of the gastrocnemius muscle in diabetic rats.
Materials and Methods: In this experimental research, 42 wistar rats (age, 8 weeks; weight, 228.31±12.1 gr) were randomly divided into seven groups of normal control (C), diabetic (D), diabetic ischemic (I), diabetic resistance training (R), diabetic endurance training (E), diabetes ischemic+resistance training (IR), diabetes ischemic+endurance training (IE). Diabetes was induced by one-step intraperitoneal injection of STZ (50 mg/kg). ICI was conducted by tightening a tourniquet (with a width of 8 mm) around the upper thighs 20 min before exercise and included three 5-min cycles of ischemia, followed by 5 min reperfusion. The training groups performed exercise training for six weeks, five days a week. Resistance training included climbing a ladder by attaching a weight to rats’ tails at 60% of maximum voluntary carrying capacity (14 repetitions with one minute rest between repetitions). Endurance training involved running on a treadmill. The treadmill speed for the first week was set at 9 meters per minute for 15 minutes. By the sixth week, the training speed was increased to 18 meters per minute for 30 min. Murf-1 and atrogin-1 gene expression were measured by RT-PCR and the diameter of fibers of gastrocnemius muscle through the staining with hematoxylin and eosin methods. Data were analyzed by using two-way analysis of variance and Bonferroni’s post-hoc tests.
Results: Murf-1 and Atrogin-1 gene expression significantly increased and fiber diameter of gastrocnemius muscle decreased in D group compared to C group (p=0.0001). Murf-1 and atrogin-1 gene expression decreased significantly in R (p=0.0001, p=0.0001), E (p=0.0001, p=0.0003), IR (p=0.0001, p=0.0001) and IE (p=0.0001, p=0.0001) compared to the C group. In addition, muscle fiber diameter increased significantly in R (p=0.034), E (p=0.007), IR (p=0.0003) and IE (p=0.0003) compared to the C.
Conclusion: Based on decreases in Murf-1 and Atrogin-1 gene expression and increases in muscle fiber diameter. It could be concluded that aerobic and resistance exercise with intermittent cycles of ischemia are more effective than any of the exercise interventions alone in preventing muscular atrophy in diabetic rats.
 
Keywords
Exercise training
Muscular Atrophy
Hyperglycemia
Ischemia
Subjects

  1. 1- Nellaiappan K, Preeti K, Khatri DK, Singh SB. Diabetic Complications: An Update on Pathobiology and Therapeutic Strategies. Curr Diabetes Rev. 2022;18:e030821192146. Doi: 10.2174/1573399817666 103091042032- O'Neill BT, Bhardwaj G, Penniman CM, Krumpoch MT, Suarez Beltran PA, Klaus K, et al. Foxo Transcription Factors are Critical Regulators of Diabetes-Related Muscle Atrophy. Diabetes. 2019;68:556–70. Doi: 10.2337/db18-04163- Ashraf JM, Ahmad S, Rabbani G, Hasan Q, Jan AT, Lee EJ, et al. 3-Deoxyglucosone: a potential glycating agent accountable for structural alteration in H3 histone protein through generation of different AGEs. PloS one. 2015;10(2):e0116804. Doi.org/10.1371/journal.pone.01168044- Sartori R, Romanello V, and Sandri M. Mechanisms of muscle atrophy and hypertrophy: Implications in health and disease. Nature Communications. 2021;12(1): 1-12. https://Doi.org/10.1038/s41467-020-20123-1

    5- Ebert SM, Al-Zougbi A, Bodine SC, Adams CM. Skeletal muscle atrophy: discovery of mechanisms and potential therapies. Physiology. 2019;34(4):232-9.

    6. Jun L, Robinson M, Geetha T, Broderick TL, Babu JR. Prevalence and Mechanisms of Skeletal Muscle Atrophy in Metabolic Conditions. Int J Mol Sci. 2023;24: 2973. https://Doi.org/10.3390/ ijms240329737- Jun L, Robinson M, Geetha T, Broderick TL, Babu JR. Prevalence and Mechanisms of Skeletal Muscle Atrophy in Metabolic Conditions. Inter J Mol Sci. 2023;24(3):2973. https://Doi.org/10.3390/ijms24032973

    1. O’Brien L, Jacobs I. Methodological variations contributing to heterogenous ergogenic responses to ischemic preconditioning. Front. Physiol. 2021;12(6), 656980. Doi:10.3389/fphys.2021.656980
    2. Lang JA, Kim J, Lang JA, Kim J. Remote ischaemic preconditioning-translating cardiovascular benefits to humans. J. Physiology. 2022;600: 3053–3067. Doi:10.1113/jp282568
    3. Jones H, Nyakayiru J, Bailey TG, Green DJ, Cable NT, Sprung VS, et al. Impact of eight weeks of repeated ischaemic preconditioning on brachial artery and cutaneous microcirculatory function in healthy males. Euro Preven Cardio. 2015;22:1083-87.DOI: 10.1177/2047487314547657
    4. Kono Y, Fukuda S, Hanatani A, Nakanishi K, Otsuka K, Taguchi H, et al. Remote ischemic conditioning improves coronary microcirculation in healthy subjects and patients with heart failure. Drug Des Devel Ther. 2014;27(8):1175-81. Doi: 10.2147/DDDT.S68715
    5. Maxwell JD, Carter HH, Hellsten Y, Miller GD, Sprung VS, Cuthbertson DJ, et al. Seven-day remote ischaemic preconditioning improves endothelial function in patients with type 2 diabetes mellitus: a randomised pilot study. Eur J Endocrinol. 2019;181(6):659-669. DOI: 10.1530/EJE-19-0378
    6. Hyngstrom AS, Murphy SA, Nguyen J, Schmit BD, Negro F, Gutterman DD, et al. Ischemic conditioning increases strength and volitional activation of paretic muscle in chronic stroke: a pilot study. J Appl Physiol. 2018;124(5):1140-7. DOI: 10.1152/japplphysiol.01072.2017
    7. Durand MJ, Boerger TF, Nguyen JN, Alqahtani SZ, Wright MT, Schmit BD, et al. Two weeks of ischemic conditioning improves walking speed and reduces neuromuscular fatigability in chronic stroke survivors. J Appl Physiol. 2019;126(3):755–63. DOI: 10.1152/japplphysiol.00772.2018

    15. Leurcharusmee P, Sawaddiruk P, Punjasawadwong Y, Sugandhavesa N, Klunklin K, Tongprasert S, et al. Ischemic preconditioning upregulates Mitofusin2 and preserves muscle strength in tourniquet-induced ischemia/reperfusion. J Orthop Translat. 2022;14:35:113-121. Doi: 10.1016/j.jot.2022.09.012. 16. Patterson SD, Swan R, Page W, Marocolo M, Jeffries O, Waldron M. The effect of acute and repeated ischemic preconditioning on recovery following exercise-induced muscle damage. J. Sci. Med. Sport. 2021;24,709-714. Doi:10.1016/j.jsams.2021.02.01217. Incognito AV, Burr JF, Millar PJ. The Effects of Ischemic Preconditioning on Human Exercise Performance. Sports Med. 2016;46(4):531–44. DOI: 10.1007/s40279-015-0433-5 18. Atherton PJ, Babraj J, Smith K, Singh J, Rennie MJ, Wackerhage H. Selective activation of AMPK-PGC-1alpha or PKB-TSC2-mTOR signaling can explain specific adaptive responses to endurance or resistance training-like electrical musclestimulation. FASEB J. 2005;19(7):786-8. Doi: 10.1096/fj.04-2179fje.

    1. Macedo AG,Oliveira DM. The Influence of the Aerobic Training on Muscle Hypertrophy: Literature Review. J health sci. 2019;21(4):382. https://Doi.org/10.17921/2447-8938.
    2. Léger B, Cartoni R, Praz M, Lamon S, Dériaz O, Crettenand A, et al. Akt signalling through GSK-3beta, mTOR and Foxo1 is involved in human skeletal muscle hypertrophy and atrophy. Journal of Physiology. 2016;576(3):923-33. DOI:10.1113/ jphysiol.2006.116715
    3. Moradi Y, Zehsaz F, Nourazar MA. Concurrent exercise training and Murf-l and Atrogin-1 gene expression in the vastus lateralis muscle of male Wistar rats. Apunts Sports Medicine, 2020;55:21-27. DOI: 10.1016/j.apunsm.2020.02.001

    22. Shanazari Z, Faramarzi M, Kordi MR. The effect of eight weeks of moderate and high intensity resistance training on miR-23a, Atrogin-1 and MuRF gene expression in fast and slow twitch muscles in Wistar older rats. Practical Studies of Biosciences in Sport. 2022;10(24),42-52. [In Persian]23. Delfan M, Bouriaei T. Synergistic Effect of 4 Weeks of Endurance Training With Probiotic Supplementation on the Expression of Atrogin-1 and Murf-1 Genes in the Soleus Muscle of Diabetic Rats. Ijdld. 2021;21(4):198-209. [In Persian]24. Cheng Z, Li L, Mo X, Zhang L, Xie Y, Guo Q, et al. Non-invasive remote limb ischemic postconditioning protects rats against focal cerebral ischemia by upregulating STAT3 and reducing apoptosis. Int J Mol Med. 2014;34(4):957-66. DOI: 10.3892/ijmm.2014.187325. Krug AL, Macedo AG, Zago AS, Rush JW, Santos CF, Amaral SL. High‐intensity resistance training attenuates dexamethasone‐induced muscle atrophy. Muscle & Nerve. 2016;53(5),779-788. DOI: 10.1002/mus.2490626. Arabzadeh E, Samadian Z, Tofighi A. Alteration of follistatin-like 1, neuron-derived neurotrophic factor, and vascular endothelial growth factor in diabetic cardiac muscle after moderate-intensity aerobic exercise with insulin. Sport Sci Health. 2020;16,491-499. DOI: 10.1007/s11332-020-00631-9.

    1. Lu F, Lu B, Zhang L, Wen J, Wang M, Zhang S, Li Q, et al. Hydrogen sulphide ameliorating skeletal muscle atrophy in db/db mice via Muscle RING finger 1 S‐sulfhydration. Cel Mole Med. 2020;24(16):9362-9377. DOI: 10.1111/jcmm.15587.
    2. Vechetti-Junior IJ, Bertaglia RS, Fernandez GJ, de Paula TG, de Souza RW, Moraes LN, et al. Aerobic exercise recovers disuse-induced atrophy through the stimulus of the LRP130/PGC-1α complex in aged rats. J Gerontol A Biol Sci Med Sci. 2016;71(5):601-609. DOI: 10.1093/gerona/glv064
    3. Sinclair AJ, Abdelhafiz AH, Rodríguez-Mañas L. Frailty and sarcopenia - newly emerging and high impact complications of diabetes. J Diabetes Complications. 2017;31(9):1465–73.
    4. Bodine SC, Baehr LM. Skeletal muscle atrophy and the E3ubiquitin ligases MuRF1 and MAFbx/atrogin-1. AmericanJournal of Physiology-Endocrinology and Metabolism. 2014;6:307:469-84. DOI:10.1152/ajpendo.00204.2014
    5. Zhang Z, Wang B, Fei A. BDNF contributes to the skeletal muscle anti-atrophic effect of exercise training through AMPK-PGC1α signaling in heart failure mice. Archives of Medical Science: AMS. 2019;15(1):214-22. DOI:10.5114/aoms.2018.81037
    6. Tanaka D, Suga T, Kido K, Honjo T, Hamaoka T, and Isaka T. Acute remote ischemic preconditioning has no effect on quadriceps muscle endurance. Transl. Sports Med. 2020;3:314-320. https://Doi.org/10.1002/tsm2.149
    7. Kubota A, Sakuraba K, Sawaki K, Sumide T, and Tamura Y. Prevention of disuse muscular weakness by restriction of blood flow. Med. Sci. Sports Exerc. 2008;40:529–534. DOI: 10.1249/MSS.0b013e31815ddac6
    8. Pryds K, Nielsen R, Jorsal A, Hansen MS, Ringgaard S, Refsgaard J, et al. Effect of long-term remote ischemic conditioning in patients with chronic ischemic heart failure. Basic Res. Cardiol. 2017;112:67. DOI: 10.1007/s00395-017-0658-6
    9. Slysz JT, Burr JF. Impact of 8 weeks of repeated ischemic preconditioning on running performance. Eur J Appl Physiol. 2019;119(6):1431-1437. DOI: 10.1007/s00421-019-04133-6
    10. Rassaf T, Totzeck M, Hendgen-Cotta UB, Shiva S, Heusch G, Kelm M. Circulating nitrite contributes to cardioprotection by remote ischemic preconditioning. Circ Res. 2014;9:114(10):1601-10. DOI: 10.1161/CIRCRESAHA.114.303822
    11. Jeffries O, Waldron M, Pattison JR, PattersonSD. Enhanced local skeletal muscle oxidative capacity and microvascular blood flow following 7-day ischemic preconditioning in healthy humans. Front. Physiol. 2018;9:463. DOI: 10.3389/fphys.2018.00463

    38. Lindsay A, Petersen C, Blackwell G, Ferguson H, Parker G, Steyn N, et al. The effect of 1 week of repeated ischaemic leg preconditioning on simulated Keirin cycling performance: a randomised trial. Sport Exerc. Med. 2017;31(3):e00229. Doi: 10.1136/bmjsem-2017-00022939. Carvalho L, Concon V, Meloni M, De Souza EO, Barroso R. Effects of resistance training combined with ischemic preconditioning on muscle size and strength in resistance-trained individuals. J Sports Med Phys Fitness. 2020;60(11):143-36. DOI: 10.23736/S0022-4707.20.11032-6.40. Surkar SM, Bland MD, Mattlage AE, Chen L, Gidday JM, Lee JM, et al. Effects of remote limb ischemic conditioning on muscle strength in healthy young adults: A randomized controlled trial. PLoS One. 2020;4:15(2):e0227263. Doi: 10.1371/journal.pone.0227263

  • Receive Date 11 May 2024
  • Revise Date 08 July 2024
  • Accept Date 13 July 2024
  • First Publish Date 13 July 2024
  • Publish Date 21 June 2024