Molecular Biology Reports

, Volume 41, Issue 11, pp 7715–7722 | Cite as

Protective effect of myokine IL-15 against H2O2-mediated oxidative stress in skeletal muscle cells

  • Fengna Li
  • Yinghui Li
  • Yulong Tang
  • Binbin Lin
  • Xiangfeng Kong
  • Oso Abimbola Oladele
  • Yulong YinEmail author


The production of reactive oxygen species (ROS) during oxidative stress may cause cellular injury. Interleukin-15 (IL-15) is one of the skeletal muscle secreted myokines, and there is no information that reported its anti-oxidative capability in skeletal muscle. The aim of this study therefore is to investigate the protective effects of myokine IL-15 against H2O2-mediated oxidative stress in C2C12 myoblasts. The results showed that IL-15 pre-incubation reduced the intracellular creatine kinase and lactate dehydrogenase activities, decreased the ROS overload, and protect the mitochondrial network via up-regulated mRNA expression levels of IL-15 and uncoupling protein 3. It also down-regulated the levels of IL-6 and p21 of the myoblasts compared to the cells treated only with H2O2. Meanwhile, apurinic/aprimidinic endonuclease 1 expression and the Akt signaling pathway were stimulated. These effects could contribute to the resumption of cell viability and act as protective mechanism. In conclusion, myokine IL-15 could be a novel endogenous regulator to control intracellular ROS production and attenuate oxidative stress in skeletal muscle cells.


IL-15 Oxidative stress H2O2 C2C12 cells 



Apurinic/aprimidinic endonuclease 1


Creatin kinase


Dulbecco’s modified Eagle’s medium


Extracellular-signal regulated kinase-1/2


Lactate dehydrogenase


Myogenic differentiation antigen


Phosphate buffered saline


Reactive oxygen species


Superoxide dismutase


Uncoupling protein 3



This study was jointly supported by National Basic Research Program of China (2013CB127305, 2012CB124704), National Nature Science Foundation of China (31372325, 31110103909), and the Project of Institute of Subtropical Agriculture, the Chinese Academy of Sciences (ISACX-LYQY-QN-1104). The authors’ contributions were as follows: F. N. Li and Y. L. Yin were in charge of the whole trial. F. N. Li wrote the manuscript while A.O Oso assisted in technical editing and correction of the paper. Y. H. Li, Y. L. Tan and B. B. Lin assisted with the cell culture. X. F. Kong assisted with the biochemical analyses.

Conflict of interest

The authors have declared that no competing interests exist.


  1. 1.
    Fan X, Hussien R, Brooks GA (2010) H2O2-induced mitochondrial fragmentation in C2C12 myocytes. Free Radic Biol Med 49(11):1646–1654PubMedCrossRefPubMedCentralGoogle Scholar
  2. 2.
    Turki A, Hayot M, Carnac G, Pillard F, Passerieux E, Bommart S, Raynaud de Mauverger E, Hugon G, Pincemail J, Pietri S, Lambert K, Belayew A, Vassetzky Y, Juntas Morales R, Mercier J, Laoudj-Chenivesse D (2012) Functional muscle impairment in facioscapulohumeral muscular dystrophy is correlated with oxidative stress and mitochondrial dysfunction. Free Radic Biol Med 53(5):1068–1079PubMedCrossRefGoogle Scholar
  3. 3.
    Febbraio MA, Pedersen BK (2005) Contraction-induced myokine production and release: is skeletal muscle an endocrine organ? Exerc Sport Sci Rev 33:114–119PubMedCrossRefGoogle Scholar
  4. 4.
    Pedersen BK, Edward F (2009) Adolph distinguished lecture: muscle as an endocrine organ: IL-6 and other myokines. J Appl Physiol 107:1006–1014PubMedCrossRefGoogle Scholar
  5. 5.
    Burton JD, Bamford RN, Peters C, Grant AJ, Kurys G, Goldman CK, Brennan J, Roessler E, Waldmann TA (1994) A lymphokine, provisionally designated as interleukin T and produced by a human adult T-cell leukemia line, stimulates T-cell proliferation and the induction of lymphokine-activated killer cells. Proc Natl Acad Sci USA 91:4935–4939PubMedCrossRefPubMedCentralGoogle Scholar
  6. 6.
    Grabstein KH, Eisenman J, Shanebeck K, Rauch C, Srinivasan S, Fung V, Beers C, Richardson J, Schoenborn MA, Ahdieh M, Johnson L, Alderson MR, Watson JD, Anderson DM, Giri JG (1994) Cloning of a T cell growth factor that interacts with the beta chain of the interleukin-2 receptor. Science 26:965–968CrossRefGoogle Scholar
  7. 7.
    Sánchez-Jiménez R, Alvarado-Vásquez N (2013) IL-15 that a regulator of TNF-α in patients with diabetes mellitus type 2. Med Hypotheses 80(6):776–777PubMedCrossRefGoogle Scholar
  8. 8.
    Busquets S, Figueras M, Almendro V, Lopez-Soriano FJ, Argiles JM (2006) Interleukin-15 increases glucose uptake in skeletal muscle. An antidiabetogenic effect of the cytokine. Biochim Biophys Acta 1760:1613–1617PubMedCrossRefGoogle Scholar
  9. 9.
    Wu X, Pan W, He Y, Hsuchou H, Kastin AJ (2010) Cerebral interleukin-15 shows upregulation and beneficial effects in experimental autoimmune encephalomyelitis. J Neuroimmunol 223(1–2):65–72PubMedCrossRefPubMedCentralGoogle Scholar
  10. 10.
    Pagliari D, Cianci R, Frosali S, Landolfi R, Cammarota G, Newton EE, Pandolfi F (2013) The role of IL-15 in gastrointestinal diseases: a bridge between innate and adaptive immune response. Cytokine Growth Factor Rev 24(5):455–466PubMedCrossRefGoogle Scholar
  11. 11.
    van der Windt GJ, Everts B, Chang CH, Curtis JD, Freitas TC, Amiel E, Pearce EJ, Pearce EL (2012) Mitochondrial respiratory capacity is a critical regulator of CD8+ T cell memory development. Immunity 36:68–78PubMedCrossRefPubMedCentralGoogle Scholar
  12. 12.
    Quinn LS, Anderson BG, Conner JD, Wolden-Hanson T (2013) IL-15 overexpression promotes endurance, oxidative energy metabolism, and muscle PPARdelta, SIRT1, PGC-1alpha, and PGC-1beta expression in male mice. Endocrinology 154(1):232–245PubMedCrossRefPubMedCentralGoogle Scholar
  13. 13.
    Li FN, Yang HS, Duan YH, Yin YL (2011) Myostatin regulates preadipocyte differentiation and lipid metabolism of adipocyte via ERK1/2. Cell Biol Int 35:1141–1146PubMedCrossRefGoogle Scholar
  14. 14.
    Barzilai A, Yamamoto K (2004) DNA damage responses to oxidative stress. DNA Repair (Amst) 3:1109–1115CrossRefGoogle Scholar
  15. 15.
    Renault V, Thornell LE, Butler-Browne G, Mouly V (2002) Human skeletal muscle satellite cells: aging, oxidative stress and the mitotic clock. Exp Gerontol 37:1229–1236PubMedCrossRefGoogle Scholar
  16. 16.
    Taylor RP, Starnes JW (2003) Age, cell signalling and cardioprotection. Acta Physiol Scand 178:107–116PubMedCrossRefGoogle Scholar
  17. 17.
    St-Pierre J, Drori S, Uldry M, Silvaggi JM, Rhee J, Jager S, Handschin C, Zheng K, Lin J, Yang W, Simon K, Bachoo R, Spiegelman BM (2006) Suppression of reactive oxygen species and neurodegeneration by the PGC-1 transcriptional coactivators. Cell 127:397–408PubMedCrossRefGoogle Scholar
  18. 18.
    Yoon JH, Kim J, Song P, Lee TG, Suh PG, Ryu SH (2012) Secretomics for skeletal muscle cells: a discovery of novel regulators? Adv Biol Regul 52(2):340–350PubMedCrossRefGoogle Scholar
  19. 19.
    Shrikant P, Mescher MF (2002) Opposing effects of IL-2 in tumor immunotherapy: promoting CD8 T cell growth and inducing apoptosis. J Immunol 169:1753–1759PubMedCrossRefGoogle Scholar
  20. 20.
    Berger C, Berger M, Hackman RC, Gough M, Elliott C, Jensen MC, Riddell SR (2009) Safety and immunologic effects of IL-15 administration in nonhuman primates. Blood 114:2417–2426PubMedCrossRefPubMedCentralGoogle Scholar
  21. 21.
    Tang C, Yamada H, Shibata K, Yoshida S, Wajjwalku W, Yoshikai Y (2009) IL-15 protects antigenspecific CD8+ T cell contraction after Mycobacterium bovis bacillus Calmette-Guerin infection. J Leukoc Biol 86:187–194PubMedCrossRefGoogle Scholar
  22. 22.
    Kosmidou I, Vassilakopoulos T, Xagorari A, Zakynthinos S, Papapetropoulos A, Roussos C (2002) Production of interleukin-6 by skeletal myotubes: role of reactive oxygen species. Am J Respir Cell Mol Biol 26(5):587–593PubMedCrossRefGoogle Scholar
  23. 23.
    Donges CE, Duffield R, Drinkwater EJ (2010) Effects of resistance or aerobic exercise training on interleukin-6, C-reactive protein, and body composition. Med Sci Sport Exer 42(2):304–313CrossRefGoogle Scholar
  24. 24.
    Pan HY, Xu XY, Hao XM, Chen YJ (2012) Changes of myogenic reactive oxygen species and interleukin-6 in contracting skeletal muscle cells. Oxid Med Cell Longev: ID145418Google Scholar
  25. 25.
    Kim JJ, Lee SB, Park JK, Yoo YD (2010) TNF-alpha-induced ROS production triggering apoptosis is directly linked to Romo1 and Bcl-X (L). Cell Death Differ 17(9):1420–1434PubMedCrossRefGoogle Scholar
  26. 26.
    Halevy O, Novitch BG, Spicer DB, Skapek SX, Rhee J, Hannon GJ, Beach D, Lassar AB (1995) Correlation of terminal cell cycle arrest of skeletal muscle with induction of p21 by MyoD. Science 267(5200):1018–1021PubMedCrossRefGoogle Scholar
  27. 27.
    Yin Y, Solomon G, Deng C, Barrett JC (1999) Differential regulation of p21 by p53 and Rb in cellular response to oxidative stress. Mol Carcinog 24:15–24PubMedCrossRefGoogle Scholar
  28. 28.
    Passos JF, Nelson G, Wang C, Richter T, Simillion C, Proctor CJ, Miwa S, Olijslagers S, Hallinan J, Wipat A, Saretzki G, Rudolph KL, Kirkwood TB, von Zglinicki T (2010) Feedback between p21 and reactive oxygen production is necessary for cell senescence. Mol Syst Biol 6:347PubMedCrossRefPubMedCentralGoogle Scholar
  29. 29.
    Gartel AL (2005) The conflicting roles of the cdk inhibitor p21CIP1/WAF1 in apoptosis. Leuk Res 29:1237–1238PubMedCrossRefGoogle Scholar
  30. 30.
    Abbas T, Dutta A (2009) p21 in cancer: intricate networks and multiple activities. Nat Rev Cancer 9:400–414PubMedCrossRefPubMedCentralGoogle Scholar
  31. 31.
    Inoue T, Kato K, Kato H, Asanoma K, Kuboyama A, Ueoka Y, Yamaguchi SI, Ohgami T, Wake N (2009) Level of reactive oxygen species induced by p21Waf1/CIP1 is critical for the determination of cell fate. Cancer Sci 100(7):1275–1283PubMedCrossRefGoogle Scholar
  32. 32.
    Azzu V, Brand MD (2010) The on-off switches of the mitochondrial uncoupling proteins. Trends Biochem Sci 35:298–307PubMedCrossRefPubMedCentralGoogle Scholar
  33. 33.
    Brand MD, Esteves TC (2005) Physiological functions of the mitochondrial uncoupling proteins UCP2 and UCP3. Cell Metab 2:85–93PubMedCrossRefGoogle Scholar
  34. 34.
    Mailloux RJ, Harper ME (2011) Uncoupling proteins and the control of mitochondrial reactive oxygen species production. Free Radic Biol Med 51:1106–1115PubMedCrossRefGoogle Scholar
  35. 35.
    Li MX, Wang D, Zhong ZY, Xiang DB, Li ZP, Xie JY, Yang ZZ, Jin F, Qing Y (2008) Targeting truncated APE1 in mitochondria enhances cell survival after oxidative stress. Free Radic Biol Med 45:592–601PubMedCrossRefGoogle Scholar
  36. 36.
    Tell G, Fantini D, Quadrifoglio F (2010) Understanding different functions of mammalian AP-endonuclease (APE1) as a promising tool for cancer treat-ment. Cell Mol Life Sci 67:3589–3608PubMedCrossRefGoogle Scholar
  37. 37.
    Lahair MM, Howe CJ, Rodriguez-Mora O, McCubrey JA, Franklin RA (2006) Molecular pathways leading to oxidative stress-induced phosphorylation of Akt. Antioxid Redox Signal 8(9–10):1749–1756PubMedCrossRefGoogle Scholar
  38. 38.
    Baar K (2004) Involvement of PPAR gamma co-activator-1, nuclear respiratory factors 1 and 2, and PPAR alpha in the adaptive response to endurance exercise. Proc Nutr Soc 63:269–273PubMedCrossRefGoogle Scholar
  39. 39.
    Sen CK, Roy S (2005) Relief from a heavy heart: redox-sensitive NF-kappaB as a therapeutic target in managing cardiac hypertrophy. Am J Physiol Heart Circ Physiol 289:H17–H19PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Fengna Li
    • 1
  • Yinghui Li
    • 1
    • 2
  • Yulong Tang
    • 1
  • Binbin Lin
    • 1
    • 3
  • Xiangfeng Kong
    • 1
  • Oso Abimbola Oladele
    • 4
  • Yulong Yin
    • 1
    Email author
  1. 1.Scientific Observing and Experimental Station of Animal Nutrition and Feed Science in South-Central, Ministry of Agriculture, Hunan Provincial Engineering Research Center of Healthy Livestock, Key Laboratory of Agro-Ecological Processes in Subtropical Region, Institute of Subtropical AgricultureChinese Academy of SciencesChangshaChina
  2. 2.University of Chinese Academy of SciencesBeijingChina
  3. 3.College of Animal SciencesHunan Agricultural UniversityChangshaChina
  4. 4.Animal Nutrition Department, College of Animal Science and Livestock ProductionFederal University of Agriculture AbeokutaAbeokutaNigeria

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