Influence of autophagy-mediated high-intensity interval training on skeletal muscle mass and aerobic capacity of middle-aged rats

doi:10.3969/j.issn.2095-4344.0137

Abstract

Abstract:

BACKGROUND: It is unclear whether autophagy mediates the long-term exercise adaptation of the skeletal muscle induced by high-intensity interval training (HIIT) .
OBJECTIVE: To identify the influence of HIIT on skeletal muscle autophagy of middle-aged rats over time, and to understand the potential regulatory effects of autophagy on maintaining the muscle mass and improving aerobic capacity by HIIT.
METHODS: Rats were randomly divided into quiet, moderate training group (50-minute running at the intensity of 60% VO2 max) and HIIT group (6 times of 3-minute running at 80% VO2 max and 3-minute active recovery at 50% VO2 max with a 7-minute warm-up and a 7-minute cool-down at 60% VO2 max). All rats were tested for VO2 max, exhaustive running time and distance at baseline and after 4, 8 and 12 weeks of exercise, respectively. The soleus muscle were collected and weighted, and then the expression levels of autophagy-related protein Beclin 1, LC3 and P62 were detected by western blot assay.
RESULTS AND CONCLUSION: The soleus muscle mass in the quiet group decreased at the 12th week (P < 0.01), while the moderate training and HIIT groups improved this decline (P < 0.05). The HIIT group dramatically improved the VO2 max from the 4th week when compared with the pre-experiment and the quiet group, until the 12th week. The expression levels of Beclin1 and LC3II in the quiet group declined at the 12th week (P < 0.05, P < 0.001), but the expression level of LC3II and ratio of LC3II/LC3I raised from the 4th and 8th weeks till the 12th week compared with the pre-experiment and the quiet group, and the expression level of LC3II and ratio of LC3II/LC3I point time significantly raised in the moderate training group at the 12th week (P < 0.001, P < 0.01). In the quiet group, the content of P62 significantly increased at the 4th and 8th weeks (P < 0.05), and decreased at the 12th week (P < 0.001), but the content of P62 remained at the low level in the moderate exercise and HIIT groups. These results imply that an equivalent or better effects on improving basic autophagy level, skeletal muscle mass and aerobic capacity of the middle aged rats, by HIIT versus moderate training, providing the theory and empirical data that highlight the role of HIIT in health promotion.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: 高强度间歇运动, 中等强度持续运动, 骨骼肌质量, 有氧运动能力, 细胞自噬, 中年大鼠, 运动适应, 组织工程

CLC Number:

R318

Cui Xin-wen, Zhang Yi-min, Wang Zan, Kong Zhen-xing. Influence of autophagy-mediated high-intensity interval training on skeletal muscle mass and aerobic capacity of middle-aged rats[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(8): 1196-1204.

Figures/Tables 2

2.1 实验动物数量分析除中等强度运动组和高强度间歇运动组个别大鼠在训练期间尾巴被跑台夹伤外，无其他大鼠受伤、感染，生病等意外情况，最终80只大鼠进入结果分析。 2.2 两种运动方式对中年大鼠骨骼肌质量随时间变化的影响采用双因素方差分析，以体质量为协变量，在体质量=600.17 g时估计两种运动方式对比目鱼肌湿质量随时间变化的影响。发现不同运动方式和时间都对比目鱼肌湿质量产生了影响。安静组大鼠比目鱼肌湿质量在干预4周时达到峰值，从8周开始略有下降，到干预12周(大鼠12月龄)时，相对于干预4周时下降了15%(P=0.003，95%CI：4%-25%)而此时中等强度运动组和高强度间歇运动组同样继续保持较高水平，分别高于同期安静组14%和12% (P=0.012，95%CI：2%-24%；P=0.014，95%CI：2%-22%)，中等强度运动组与高强度间歇运动组无差异，见图1。高强度间歇运动和中等强度运动均有效抑制了12月龄大鼠比目鱼肌湿质量的下降，但两种运动方式对比目鱼肌湿质量的影响无明显差异。 2.3 两种运动方式对中年大鼠有氧运动能力随时间变化的影响双因素方差分析两种运动方式对最大摄氧量和力竭运动时间和力竭运动距离随时间变化的影响，发现不同运动方式和时间都对最大摄氧量和力竭运动时间和力竭运动距离产生了影响。在对最大摄氧量的影响中，干预4周时高强度间歇运动组相对于干预前提高了25%(P=0.002，95%CI：7%-42%)，相对于安静组提高了26%(P=0.000，95%CI：10%-43%)，相对于中等强度运动组提高了16%(P=0.031，95%CI：1%-31%)；干预8周时高强度间歇运动组相对于干预前提高了30%(P=0.001，95%CI：10%-50%；P=0.000，95%CI：13%-43%)，相对于安静组提高了22%(P=0.013，95%CI：3.6%-40.6%)；12周时高强度间歇运动组相对于干预前提高了40%(P=0.000，95%CI：15%-65%)，相对于安静组提高了26%(P=0.034，95%CI：1.4%-51.5%)，见图2。安静组在整个干预过程中无明显波动，中等强度运动组从干预第4周开始有上升趋势，但差异无显著性意义，而高强度间歇运动组从干预第4周时开始，最大摄氧量显著提高，12周时达到最高水平，可知，高强度间歇运动较中等强度运动明显提高了最大摄氧量。在对力竭运动时间的影响中，4周时中等强度运动组和高强度间歇运动组相对于干预前分别提高了17%和23%(P=0.006，95%CI：4%-30%；P=0.000，95%CI：10%-30%)，相对于安静组分别提高了18%和27% (P=0.001，95%CI：6%-31%；P=0.000，95%CI：15%- 27%)，但二者无显著性差异；8周时中等强度运动组和高强度间歇运动组相对于干预前分别提高了22%和28% (P=0.001，95% CI：7%-37%)，P=0.000，95%CI：13%-43%)，相对于安静组分别提高了31%和40% (P=0.000，95%CI：15%-47%；P=0.000，95%CI：24%- 56%)，但二者无显著性差异；干预12周中等强度运动组和高强度间歇运动组相对于干预前分别提高了45%和46% (P=0.001，95%CI：26%-64%，P=0.000，95%CI：27%- 64%)，相对于安静组分别提高了55%和57% (P=0.000，95%CI：32%-77%；P=0.000，95%CI：35%-80%)，但二者无显著性差异，见图3。可见，安静组在整个干预过程中没有明显波动，而中等强度运动组和高强度间歇运动组从干预4周时开始，力竭运动时间逐步提高，直到12周时达到最高水平，分别超出对照组55%和57%，但两种运动方式在提高力竭运动时间上无明显的差异。在对力竭运动距离的影响中，干预4周中等强度运动组和高强度间歇运动组相对于干预前分别提高了35%和48% (P=0.014，95%CI：5%-64%；P=0.000，95%CI：20%-76%)，相对于安静组分别提高了38%和58% (P=0.001，95%CI：10%-66%；P=0.000，95%CI：30%-85%)，高强度间歇运动组有高于中等强度运动组的趋势(P=0.242)；8周时中等强度运动组和高强度间歇运动组相对于干预前分别提高了45%和60%(P=0.002，95%CI：12%-78%)，P=0.000，95%CI：27%-91%)，相对于安静组分别提高了64%和87%(P=0.000，95%CI：27%-101%；P=0.000，95%CI：50%-124%)，但二者无显著性差异；12周时中等强度运动组和高强度间歇运动组相对于干预前分别提高了106%和105%(P=0.001，95%CI：64%- 148%，P=0.000，95%CI：65%-146%)，相对于安静组分别提高了135%和144%(P=0.000，95%CI：82%-188%；P=0.000，95%CI：90%-197%)，但二者无显著性差异，见图4。可见，安静组在整个干预过程中没有明显波动，而中等强度运动组和高强度间歇运动组从干预4周时开始，力竭运动距离逐步提高，直到12周时达到最高水平，分别超出对照组135%和144%，且高强度间歇运动在提高力竭运动距离上有优于中等强度运动的趋势。 2.4 两种运动方式对中年大鼠骨骼肌Beclin 1、LC3和P62蛋白表达随时间变化的影响采用双因素方差分析两种运动方式对比目鱼肌Beclin 1、LC3II的蛋白表达，LC3II/LC3I比值和P62蛋白表达随时间变化的影响。在对比目鱼肌Beclin 1的蛋白表达的影响中，可见12周时安静组相对于干预前有明显的下降趋势(P=0.025)，而两个运动组略高于安静组，见图5。在对比目鱼肌对LC3II蛋白表达的影响中，干预4周时，高强度间歇运动组LC3II蛋白表达显著高于干预前和同期的安静组，分别提升了47%和28%(P=0.003，95%CI：12%-82%；P=0.048，95%CI：0%-55%)；8周时，高强度间歇运动组LC3II蛋白表达继续提升，显著高于干预前和同期的安静组，分别提升了60%和25%(P=0.000，95%CI：24%-94%；P=0.05，95%CI：0%-50%)，此时中等强度运动组也显著高于干预前40%(P=0.015，95%CI：5%-75%)，安静组相对于干预前和4周有提升的趋势，但是无显著变化；12周时，安静组LC3II蛋白表相对于4周和8周时期显著下降33%和40%(P=0.025，95%CI：3%-64%；P=0.001，95%CI：12%-67%)，而同期的中等强度运动组和高强度间歇运动组则在8周的基础上继续保持较高水平，分别高于安静组88%和78%(P=0.000，95%CI：48%- 130%；P=0.000，95%CI：38%-120%)，见图6。可见，随时间的变化，安静组LC3II蛋白表达在干预12周(大鼠12月龄)出现显著降低。高强度间歇运动组则从干预4周时起显著提升，直到实验结束，而中等强度运动组的提升相对滞后，说明两种运动方式有效促进了LC3II蛋白表达，缓解了安静组的下降趋势，且高强度间歇运动较中等强度运动更具有时效性。"

References

[1] Sandri M. Autophagy in health and disease. 3. Involvement of autophagy in muscle atrophy. Am J Physiol Cell Physiol. 2010;298(6): C1291-C1297.

[2] Masiero E, Agatea L, Mammucari C, et al. Autophagy is required to maintain muscle mass. Cell Metab. 2009;10(6):507-515.

[3] Papackova Z, Cahova M. Important role of autophagy in regulation of metabolic processes in health, disease and aging. Physiol Res. 2014; 63(4):409-420.

[4] Tachtsis B, Smiles WJ, Lane SC, et al. Acute Endurance Exercise Induces Nuclear p53 Abundance in Human Skeletal Muscle. Front Physiol. 2016;7:144.

[5] Pigna E, Berardi E, Aulino P, et al. Aerobic Exercise and Pharmacological Treatments Counteract Cachexia by Modulating Autophagy in Colon Cancer. Sci Rep. 2016;6:26991.

[6] Kim JS, Lee YH, Yi HK. Gradual downhill running improves age-related skeletal muscle and bone weakness: implication of autophagy and bone morphogenetic proteins. Exp Physiol. 2016;101(12):1528-1540.

[7] Vainshtein A, Hood DA. The regulation of autophagy during exercise in skeletal muscle. J Appl Physiol (1985). 2016;120(6):664-673.

[8] White Z, Terrill J, White RB, et al. Voluntary resistance wheel exercise from mid-life prevents sarcopenia and increases markers of mitochondrial function and autophagy in muscles of old male and female C57BL/6J mice. Skelet Muscle. 2016;6(1):45.

[9] 黎涌明.高强度间歇训练对不同训练人群的应用效果[J].体育科学, 2015, 35(8):59-75.

[10] Billat L V. Interval training for performance: a scientific and empirical practice. Special recommendations for middle- and long-distance running. Part I: aerobic interval training. Sports Med. 2001;31(1):13-31.

[11] Nybo L, Sundstrup E, Jakobsen MD, et al. High-intensity training versus traditional exercise interventions for promoting health. Med Sci Sports Exerc. 2010;42(10):1951-1958.

[12] Gibala MJ, Mcgee SL. Metabolic adaptations to short-term high-intensity interval training: a little pain for a lot of gain? Exerc Sport Sci Rev. 2008;36(2):58-63.

[13] 王京京,韩涵,张海峰.高强度间歇训练对青年肥胖女性腹部脂肪含量的影响[J].中国运动医学杂志,2015,34(1):15-20.

[14] Heinrich KM, Patel PM, O'neal JL, et al. High-intensity compared to moderate-intensity training for exercise initiation, enjoyment, adherence, and intentions: an intervention study. BMC Public Health. 2014;14:789.

[15] Bartlett JD, Close GL, Maclaren DP, et al. High-intensity interval running is perceived to be more enjoyable than moderate-intensity continuous exercise: implications for exercise adherence. J Sports Sci. 2011;29(6):547-553.

[16] Tjonna AE, Lee SJ, Rognmo O, et al. Aerobic interval training versus continuous moderate exercise as a treatment for the metabolic syndrome: a pilot study. Circulation. 2008;118(4):346-354.

[17] Burgomaster KA, Howarth KR, Phillips SM, et al. Similar metabolic adaptations during exercise after low volume sprint interval and traditional endurance training in humans. J Physiol. 2008;586(1): 151-160.

[18] Holloway TM, Bloemberg D, Da Silva ML, et al. High intensity interval and endurance training have opposing effects on markers of heart failure and cardiac remodeling in hypertensive rats. PLoS One. 2015; 10(3):e0121138.

[19] Bedford TG, Tipton CM, Wilson NC, et al. Maximum oxygen consumption of rats and its changes with various experimental procedures. J Appl Physiol Respir Environ Exerc Physiol. 1979;47(6): 1278-1283.

[20] Angadi SS, Mookadam F, Lee CD, et al. High-intensity interval training vs. moderate-intensity continuous exercise training in heart failure with preserved ejection fraction: a pilot study. J Appl Physiol (1985). 2015; 119(6):753-758.

[21] Smart NA, Dieberg G, Giallauria F. Intermittent versus continuous exercise training in chronic heart failure: a meta-analysis. Int J Cardiol. 2013;166(2):352-358.

[22] Arena R, Myers J, Forman DE, et al. Should high-intensity-aerobic interval training become the clinical standard in heart failure?Heart Fail Rev. 2013;18(1):95-105.

[23] Leandro CG, Levada AC, Hirabara SM, et al. A program of moderate physical training for Wistar rats based on maximal oxygen consumption. J Strength Cond Res. 2007;21(3):751-756.

[24] He C, Bassik M C, Moresi V, et al. Exercise-induced BCL2-regulated autophagy is required for muscle glucose homeostasis. Nature. 2012;481(7382):511-515.

[25] Roubenoff R. Physical activity, inflammation, and muscle loss. Nutr Rev. 2007;65(12 Pt 2):S208-S212.

[26] Betik AC, Baker DJ, Krause DJ, et al. Exercise training in late middle-aged male Fischer 344 x Brown Norway F1-hybrid rats improves skeletal muscle aerobic function. Exp Physiol. 2008; 93(7):863-871.

[27] Kim YA, Kim YS, Oh SL, et al. Autophagic response to exercise training in skeletal muscle with age. J Physiol Biochem. 2013; 69(4):697-705.

[28] Stoggl TL, Bjorklund G. High Intensity Interval Training Leads to Greater Improvements in Acute Heart Rate Recovery and Anaerobic Power as High Volume Low Intensity Training. Front Physiol. 2017;8: 562.

[29] Moholdt T, Aamot I L, Granoien I, et al. Aerobic interval training increases peak oxygen uptake more than usual care exercise training in myocardial infarction patients: a randomized controlled study. Clin Rehabil, 2012;26(1):33-44.

[30] Freyssin C, Verkindt C, Prieur F, et al. Cardiac rehabilitation in chronic heart failure: effect of an 8-week, high-intensity interval training versus continuous training. Arch Phys Med Rehabil. 2012;93(8):1359-1364.

[31] Scharf M, Schmid A, Kemmler W, et al. Myocardial adaptation to high-intensity (interval) training in previously untrained men with a longitudinal cardiovascular magnetic resonance imaging study (Running Study and Heart Trial). Circ Cardiovasc Imaging. 2015; 8(4):e002566.

[32] Metcalfe RS, Babraj JA, Fawkner SG, et al. Towards the minimal amount of exercise for improving metabolic health: beneficial effects of reduced-exertion high-intensity interval training. Eur J Appl Physiol. 2012;112(7):2767-2775.

[33] Cocks M, Shaw CS, Shepherd SO, et al. Sprint interval and endurance training are equally effective in increasing muscle microvascular density and eNOS content in sedentary males. J Physiol. 2013;591(3): 641-656.

[34] Little JP, Safdar A, Bishop D, et al. An acute bout of high-intensity interval training increases the nuclear abundance of PGC-1alpha and activates mitochondrial biogenesis in human skeletal muscle. Am J Physiol Regul Integr Comp Physiol. 2011;300(6):R1303-R1310.

[35] Bartlett JD, Hwa Joo C, Jeong TS, et al. Matched work high-intensity interval and continuous running induce similar increases in PGC-1alpha mRNA, AMPK, p38, and p53 phosphorylation in human skeletal muscle. J Appl Physiol (1985). 2012;112(7):1135-1143.

[36] Taylor CW, Ingham SA, Ferguson RA. Acute and chronic effect of sprint interval training combined with postexercise blood-flow restriction in trained individuals. Exp Physiol. 2016;101(1):143-154.

[37] Robinson MM, Dasari S, Konopka AR, et al. Enhanced Protein Translation Underlies Improved Metabolic and Physical Adaptations to Different Exercise Training Modes in Young and Old Humans. Cell Metab. 2017;25(3):581-592.

[38] Scribbans TD, Edgett BA, Vorobej K, et al. Fibre-specific responses to endurance and low volume high intensity interval training: striking similarities in acute and chronic adaptation. PLoS One. 2014;9(6): e98119.

[39] Mueller SM, Aguayo D, Zuercher M, et al. High-intensity interval training with vibration as rest intervals attenuates fiber atrophy and prevents decreases in anaerobic performance. PLoS One. 2015;10(2): e0116764.

[40] Burgomaster KA, Cermak NM, Phillips SM, et al. Divergent response of metabolite transport proteins in human skeletal muscle after sprint interval training and detraining. Am J Physiol Regul Integr Comp Physiol. 2007;292(5):R1970-R1976.

[41] Burgomaster KA, Heigenhauser GJ, Gibala MJ. Effect of short-term sprint interval training on human skeletal muscle carbohydrate metabolism during exercise and time-trial performance. J Appl Physiol (1985). 2006;100(6):2041-2047.

[42] Kim YA, Kim YS, Song W. Autophagic response to a single bout of moderate exercise in murine skeletal muscle. J Physiol Biochem. 2012;68(2):229-235.

[43] Levine B, Kroemer G. Autophagy in the pathogenesis of disease. Cell. 2008;132(1):27-42.

[44] 刘文锋,魏霞,张冰侠,等.规律运动对增龄大鼠比目鱼肌细胞凋亡与自噬及PGC-1α信号调控的影响[J].中国运动医学杂志,2017,36(2):128-135.

[45] 马晓雯,常芸,王世强,等.不同强度不同时间耐力训练对于大鼠心肌细胞自噬发生程度的影响[J].中国运动医学杂志,2016,35(1):27-31.

[46] Klionsky DJ. Autophagy: from phenomenology to molecular understanding in less than a decade. Nat Rev Mol Cell Biol. 2007;8(11): 931-937.

[47] Wirawan E, Lippens S, Vanden Berghe T, et al. Beclin1: a role in membrane dynamics and beyond. Autophagy. 2012;8(1):6-17.

[48] Lira VA, Okutsu M, Zhang M, et al. Autophagy is required for exercise training-induced skeletal muscle adaptation and improvement of physical performance. FASEB J. 2013;27(10):4184-4193.

[49] Kim J, Kundu M, Viollet B, et al. AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nat Cell Biol. 2011;13(2): 132-141.

[50] Zhao J, Brault JJ, Schild A, et al. Coordinate activation of autophagy and the proteasome pathway by FoxO transcription factor. Autophagy. 2008;4(3):378-380.

[51] Zhao J, Brault JJ, Schild A, et al. FoxO3 coordinately activates protein degradation by the autophagic/lysosomal and proteasomal pathways in atrophying muscle cells. Cell Metab. 2007;6(6):472-483.

[52] Mizushima N, Yoshimori T. How to interpret LC3 immunoblotting. Autophagy. 2007;3(6):542-545.

[53] Kabeya Y, Mizushima N, Ueno T, et al. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J. 2000;19(21):5720-5728.

[54] Kabeya Y, Mizushima N, Yamamoto A, et al. LC3, GABARAP and GATE16 localize to autophagosomal membrane depending on form-II formation. J Cell Sci. 2004;117(Pt 13):2805-2812.