, Volume 18, Issue 1, pp 85–95 | Cite as

l-Lysine suppresses myofibrillar protein degradation and autophagy in skeletal muscles of senescence-accelerated mouse prone 8

  • Tomonori SatoEmail author
  • Yoshiaki Ito
  • Takashi Nagasawa
Research Article


Sarcopenia is a condition of the loss of muscle mass that is associated with aging and that increases the risk for bedridden state, thereby warranting studies of interventions that attenuate sarcopenia. Here the effects of 2-month dietary l-lysine (Lys) supplementation (1.5–3.0 %) on myofibrillar protein degradation and major proteolytic systems were investigated in senescence-accelerated mouse prone 8 (SAMP8). At 36 weeks of age, skeletal muscle and lean body mass was reduced in SAMP8 when compared with control senescence-accelerated mouse resistant 1 (SAMR1). The myofibrillar protein degradation, which was evaluated by the release of 3-methylhistidine, was stimulated in SAMP8, and the autophagy activity, which was evaluated by light chain 3-II, was stimulated in the skeletal muscle of SAMP8. The activation of ubiquitin-proteasome system was not observed in the muscles of SAMP8. However, myofibrillar protein degradation and autophagic activity in skeletal muscles of SAMP8 were suppressed by dietary intake of 3.0 % Lys. The present data indicate that myofibrillar protein degradation by bulk autophagy is stimulated in the skeletal muscles of SAMP8 and that dietary Lys supplementation attenuates sarcopenia in SAMP8 by suppressing autophagic proteolysis.


Sarcopenia Autophagy Ubiquitin proteasome system Skeletal muscle Lysine 



Glyceraldehyde-3-phosphate dehydrogenase


Light chain 3






Mammalian target of rapamycin


Mitochondrial E3 ubiquitin protein ligase 1


Muscle ring-finger protein 1


Peroxisome proliferator activated receptor γ co-activator 1α


Quantitative reverse transcription PCR


Senescence-accelerated mouse prone 8


Senescence-accelerated mouse resistant 1


p70 ribosomal protein S6 kinase 1


Ubiquitin-proteasome system


Eukaryotic initiation factor 4E binding protein 1



Amino acids were provided by Ajinomoto Co., Inc. Primer sequences for atrogin-1 and MuRF1 were provided by Professor Takeshi Nikawa, Tokushima University, Japan.


This work was supported by the Sasakawa Scientific Research Grant from The Japan Science Society (Grant No. 27-414).

Compliance with ethical standards

Conflict of interest

We declare no conflict of interest related to this study.

Informed consent

Human subject is not involved in this study.

Research involving Animals

Male SAMP8 (15-week-old, 27–35 g, n = 17) and SAMR1 (15-week-old, 31–35 g, n = 6) were purchased from Japan SLC, Inc. (Shizuoka, Japan). All animal protocols were approved by the Iwate University Animal Research Committee and were performed in compliance with the Guidelines for Animal Experiments of Iwate University (approval number, A201517).


  1. Altun M, Besche HC, Overkleeft HS, Piccirillo R, Edelmann MJ, Kessler BM, Goldberg AL, Ulfhake B (2010) Muscle wasting in aged, sarcopenic rats is associated with enhanced activity of the ubiquitin proteasome pathway. J Biol Chem 285:39597–395608CrossRefPubMedPubMedCentralGoogle Scholar
  2. Angcajas AB, Hirai N, Kaneshiro K, Karim MR, Horii Y, Kubota M, Fujimura S, Kadowaki M (2014) Diversity of amino acid signaling pathways on autophagy regulation: a novel pathway for arginine. Biochem Biophys Res Commun 446:8–14CrossRefPubMedGoogle Scholar
  3. Anthony JC, Yoshizawa F, Anthony TG, Vary TC, Jefferson LS, Kimball SR (2000) Leucine stimulates translation initiation in skeletal muscle of post absorptive rats via a rapamycin-sensitive pathway. J Nutr 130:2413–2419PubMedGoogle Scholar
  4. Børsheim E, Bui QU, Tissier S, Kobayashi H, Ferrando AA, Wolfe RR (2008) Effect of amino acid supplementation on muscle mass, strength and physical function in elderly. Clin Nutr 27:189–195CrossRefPubMedPubMedCentralGoogle Scholar
  5. Chalil S, Pierre N, Bakker AD, Manders RJ, Pletsers A, Francaux M, Klein-Nulend J, Jaspers RT, Deldicque L (2015) Aging related ER stress is not responsible for anabolic resistance in mouse skeletal muscle. Biochem Biophys Res Commun 468:702–707CrossRefPubMedGoogle Scholar
  6. Cruz-Jentoft AJ, Baeyens JP, Bauer JM, Boirie Y, Cederholm T, Landi F, Martin FC, Michel JP, Rolland Y, Schneider SM, Topinková E, Vandewoude M, Zamboni M (2010) Sarcopenia: European consensus on definition and diagnosis: report of the European working group on sarcopenia in older people. Age Ageing 39:412–423CrossRefPubMedPubMedCentralGoogle Scholar
  7. Cuthbertson D, Smith K, Babraj J, Leese G, Waddell T, Atherton P, Wackerhage H, Taylor PM, Rennie MJ (2005) Anabolic signaling deficits underlie amino acid resistance of wasting, aging muscle. FASEB J 19:422–424PubMedGoogle Scholar
  8. Derave W, Eijnde BO, Ramaekers M, Hespel P (2005) Soleus muscles of SAMP8 mice provide an accelerated model of skeletal muscle senescence. Exp Gerontol 40:562–572CrossRefPubMedGoogle Scholar
  9. Dong W, Quo W, Wang F, Li C, Xie Y, Zheng X, Shi H (2015) Electroacupuncture upregulates SIRT1-dependent PGC-1α expression in SAMP8 mice. Med Sci Monit 21:3356–3362CrossRefPubMedPubMedCentralGoogle Scholar
  10. Edström E, Altun M, Hägglund M, Ulfhake B (2006) Atrogin-1/MAFbx and MuRF1 are downregulated in aging-related loss of skeletal muscle. J Gerontol Ser A 61:663–674CrossRefGoogle Scholar
  11. Fan J, Kou X, Jia S, Yang X, Yang Y, Chen N (2016) Autophagy as a potential target for sarcopenia. J Cell Physiol 231:1450–1459CrossRefPubMedGoogle Scholar
  12. Foletta VC, White LJ, Larsen AE, Léger B, Russell AP (2011) The role and regulation of MAFbx/atrogin-1 and MuRF1 in skeletal muscle atrophy. Pflugers Arch 461:325–335CrossRefPubMedGoogle Scholar
  13. García-Prat L, Martínez-Vicente M, Perdiguero E, Ortet L, Rodríguez-Ubreva J, Rebollo E, Ruiz-Bonilla V, Gutarra S, Ballestar E, Serrano AL, Sandri M, Muñoz-Cánoves P (2016) Autophagy maintains stemness by preventing senescence. Nature 529:37–42CrossRefPubMedGoogle Scholar
  14. Ghosh S, Lertwattanarak R, Lefort N, Molina-Carrion M, Joya-Galeana J, Bowen BP, Garduno-Garcia Jde J, Abdul-Ghani M, Richardson A, DeFronzo RA, Mandarino L, Van Remmen H, Musi N (2011) Reduction in reactive oxygen species production by mitochondria from elderly subjects with normal and impaired glucose tolerance. Diabetes 60:2051–2060CrossRefPubMedPubMedCentralGoogle Scholar
  15. Guo AY, Leung KS, Siu PM, Qin JH, Chow SK, Qin L, Li CY, Cheung WH (2015) Muscle mass, structural and functional investigations of senescence-accelerated mouse P8 (SAMP8). Exp Anim 64:425–433CrossRefPubMedPubMedCentralGoogle Scholar
  16. Hatazawa Y, Senoo N, Tadaishi M, Ogawa Y, Ezaki O, Kamei Y, Miura S (2015) Metabolomic analysis of the skeletal muscle of mice overexpressing PGC-1α. PLoS ONE 10:e0129084CrossRefPubMedPubMedCentralGoogle Scholar
  17. Hughes DC, Stewart CE, Sculthorpe N, Dugdale HF, Yousefian F, Lewis MP, Sharples AP (2016) Testosterone enables growth and hypertrophy in fusion impaired myoblasts that display myotube atrophy: deciphering the role of androgen and IGF-I receptors. Biogerontology 17:619–639CrossRefPubMedGoogle Scholar
  18. Itakura E, Kishi C, Inoue K, Mizushima N (2008) Beclin 1 forms two distinct phosphatidylinositol 3-kinase complexes with mammalian Atg14 and UVRAG. Mol Biol Cell 19:5360–5372CrossRefPubMedPubMedCentralGoogle Scholar
  19. Johansen T, Lamark T (2011) Selective autophagy mediated by autophagic adapter proteins. Autophagy 7:279–296CrossRefPubMedPubMedCentralGoogle Scholar
  20. Joseph AM, Adhihetty PJ, Buford TW, Wohlgemuth SE, Lees HA, Nguyen LM, Aranda JM, Sandesara BD, Pahor M, Manini TM, Marzetti E, Leeuwenburgh C (2012) The impact of aging on mitochondrial function and biogenesis pathways in skeletal muscle of sedentary high- and low-functioning elderly individuals. Aging Cell 11:801–809CrossRefPubMedPubMedCentralGoogle Scholar
  21. Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T (2000) LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J 19:5720–5728CrossRefPubMedPubMedCentralGoogle Scholar
  22. Kirkin V, McEwan DG, Novak I, Dikic I (2009) A role for ubiquitin in selective autophagy. Mol Cell 34:259–269CrossRefPubMedGoogle Scholar
  23. Komatsu M, Ichimura Y (2010) Physiological significance of selective degradation of p62 by autophagy. FEBS Lett 584:1374–1378CrossRefPubMedGoogle Scholar
  24. Lamont LS, McCullough AJ, Kalhan SC (2001) Gender differences in leucine, but not lysine, kinetics. J Appl Physiol 91:357–362PubMedGoogle Scholar
  25. Liu HW, Chan YC, Wang MF, Wei CC, Chang SJ (2015) Dietary (-)-epigallocatechin-3-gallate supplementation counteracts aging-associated skeletal muscle insulin resistance and fatty liver in senescence-accelerated mouse. J Agric Food Chem 63:8407–8417CrossRefPubMedGoogle Scholar
  26. Mizushima N (2007) Autophagy: process and function. Genes Dev 21:2861–2873CrossRefPubMedGoogle Scholar
  27. Nagasawa T, Hirano J, Yoshizawa F, Nishizawa N (1998) Myofibrillar protein catabolism is rapidly suppressed following protein feeding. Biosci Biotechnol Biochem 62:1932–1937CrossRefPubMedGoogle Scholar
  28. Narici MV, Maffulli N (2010) Sarcopenia: characteristics, mechanisms and functional significance. Br Med Bull 95:139–159CrossRefPubMedGoogle Scholar
  29. Pagano TB, Wojcik S, Costagliola A, De Biase D, Iovino S, Iovane V, Russo V, Papparella S, Paciello O (2015) Age related skeletal muscle atrophy and upregulation of autophagy in dogs. Vet J 206:54–60CrossRefPubMedGoogle Scholar
  30. Prado CM, Wells JC, Smith SR, Stephan BC, Siervo M (2012) Sarcopenic obesity: a critical appraisal of the current evidence. Clin Nutr 31:583–601CrossRefPubMedGoogle Scholar
  31. Reeves PG, Nielsen FH, Fahey GC Jr (1993) AIN-93 purified diets for laboratory rodents: final report of the American Institute of Nutrition ad hoc writing committee on the reformulation of the AIN-76A rodent diet. J Nutr 123:1939–1951Google Scholar
  32. Russ DW, Boyd IM, McCoy KM, McCorkle KW (2016) Muscle-specificity of age-related changes in markers of autophagy and sphingolipid metabolism. Biogerontology 16:747–759CrossRefGoogle Scholar
  33. Sakuma K, Kinoshita M, Ito Y, Aizawa M, Aoi W, Yamaguchi A (2016) p62/SQSTM1 but not LC3 is accumulated in sarcopenic muscle of mice. J Cachexia Sarcopenia Muscle 7:204–212CrossRefPubMedGoogle Scholar
  34. Sandri M, Lin J, Handschin C, Yang W, Arany ZP, Lecker SH, Goldberg AL, Spiegelman BM (2006) PGC-1alpha protects skeletal muscle from atrophy by suppressing FoxO3 action and atrophy-specific gene transcription. Proc Natl Acad Sci USA 103:16260–16265CrossRefPubMedPubMedCentralGoogle Scholar
  35. Sandri M, Barberi L, Bijlsma AY, Blaauw B, Dyar KA, Milan G, Mammucari C, Meskers CG, Pallafacchina G, Paoli A, Pion D, Roceri M, Romanello V, Serrano AL, Toniolo L, Larsson L, Maier AB, Muñoz-Cánoves P, Musarò A, Pende M, Reggiani C, Rizzuto R, Schiaffino S (2013) Signalling pathways regulating muscle mass in ageing skeletal muscle: the role of the IGF1-Akt-mTOR-FoxO pathway. Biogerontology 14:303–323CrossRefPubMedGoogle Scholar
  36. Sato T, Ito Y, Nagasawa T (2013) Regulation of skeletal muscle protein degradation and synthesis by oral administration of lysine in rats. J Nutr Sci Vitaminol 59:412–419CrossRefPubMedGoogle Scholar
  37. Sato T, Ito Y, Nagasawa T (2014) Lysine suppresses myofibrillar protein degradation by regulating the autophagic-lysosomal system through phosphorylation of Akt in C2C12 cells. Springerplus 3:584CrossRefPubMedPubMedCentralGoogle Scholar
  38. Sato T, Ito Y, Nagasawa T (2015) Dietary l-lysine suppresses autophagic proteolysis and stimulates Akt/mTOR signaling in the skeletal muscle of rats fed a low-protein diet. J Agric Food Chem 63:8192–8198CrossRefPubMedGoogle Scholar
  39. Sin TK, Yu AP, Yung BY, Yip SP, Chan LW, Wong CS, Rudd JA, Siu PM (2015) Effects of long-term resveratrol-induced SIRT1 activation on insulin and apoptotic signalling in aged skeletal muscle. Acta Diabetol 52:1063–1075CrossRefPubMedGoogle Scholar
  40. Sugawara T, Ito Y, Nishizawa N, Nagasawa T (2009) Regulation of muscle protein degradation, not synthesis, by dietary leucine in rats fed a protein-deficient diet. Amino Acids 37:609–616CrossRefPubMedGoogle Scholar
  41. Takeda T (1999) Senescence-accelerated mouse (SAM): a biogerontological resource in aging research. Neurobiol Aging 20:105–110CrossRefPubMedGoogle Scholar
  42. Thrower JS, Hoffman L, Rechsteiner M, Pickart CM (2000) Recognition of the polyubiquitin proteolytic signal. EMBO J 19:94–102CrossRefPubMedPubMedCentralGoogle Scholar
  43. Wenz T, Rossi SG, Rotundo RL, Spiegelman BM, Moraes CT (2009) Increased muscle PGC-1alpha expression protects from sarcopenia and metabolic disease during aging. Proc Natl Acad Sci USA 106:20405–20410CrossRefPubMedPubMedCentralGoogle Scholar
  44. White Z, White RB, McMahon C, Grounds MD, Shavlakadze T (2016) High mTORC1 signaling is maintained, while protein degradation pathways are perturbed in old murine skeletal muscles in the fasted state. Int J Biochem Cell Biol 78:10–21CrossRefPubMedGoogle Scholar
  45. Young VR, Munro HN (1978) Nτ–methylhistidine (3-methylhistidine) and muscle protein turnover: an overview. Fed Proc 37:2291–3000PubMedGoogle Scholar
  46. Yun J, Puri R, Yang H, Lizzio MA, Wu C, Sheng ZH, Guo M (2014) MUL1 acts in parallel to the PINK1/parkin pathway in regulating mitofusin and compensates for loss of PINK1/parkin. Elife 3:e01958CrossRefPubMedPubMedCentralGoogle Scholar
  47. Zhao J, Brault JJ, Schild A, Cao P, Sandri M, Schiaffino S, Lecker SH, Goldberg AL (2007) FoxO3 coordinately activates protein degradation by the autophagic/lysosomal and proteasomal pathways in atrophying muscle cells. Cell Metab 6:472–483CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  1. 1.Department of Bioresources Science, The United Graduate School of Agricultural SciencesIwate UniversityMoriokaJapan
  2. 2.Department of Biological Chemistry and Food Science, Graduate School of AgricultureIwate UniversityMoriokaJapan

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