Molecular and Cellular Biochemistry

, Volume 444, Issue 1–2, pp 17–25 | Cite as

Tomatidine inhibits tumor necrosis factor-α-induced apoptosis in C2C12 myoblasts via ameliorating endoplasmic reticulum stress

  • Seung-Eun Song
  • Su-Kyung Shin
  • Hyun-Woo Cho
  • Seung-Soon Im
  • Jae-Hoon Bae
  • Seon Min Woo
  • Taeg-Kyu Kwon
  • Dae-Kyu Song


In this study, we examined the effect of tomatidine on tumor necrosis factor (TNF)-α-induced apoptosis in C2C12 myoblasts. TNF-α treatment increased cleaved caspase 3 and cleaved poly (ADP-ribose) polymerase (PARP) protein levels in a dose- and time-dependent manner. Pretreatment of cells with 10 μM tomatidine prevented TNF-α-induced apoptosis, caspase 3 cleavage, and PARP cleavage. Cells were treated with 100 ng/mL TNF-α for 24 h, and flow cytometry was utilized to assess apoptosis using annexin-V and 7-aminoactinomycin D. TNF-α up-regulated activating transcription factor 4 (ATF4) and C/EBP homologous protein (CHOP) expression. This effect was suppressed by pretreatment with tomatidine. Pretreatment with 4-phenylbutyric acid (a chemical chaperone) also inhibited TNF-α-induced cleavage of caspase 3 and PARP and up-regulation of ATF4 and CHOP expression. In addition, tomatidine-mediated inhibition of phosphorylation of c-Jun amino terminal kinase (JNK) attenuated TNF-α-induced cleavage of PARP and caspase 3. However, tomatidine did not affect NF-κB activation in TNF-α-treated C2C12 myoblast cells. Taken together, the present study demonstrates that tomatidine attenuates TNF-α-induced apoptosis through down-regulation of CHOP expression and inhibition of JNK activation.


Tomatidine TNF-α C2C12 myoblast ER stress Apoptosis 



This study was funded by National Research Foundation of Korea (NRF) grants funded by the Korean Government (MSIP; Nos. 2014R1A5A2010008, 2013R1A2A2A01068220), and partially by iPET (Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries), Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea [Grant Number 110135-3]. The authors appreciate Dr. Choi BK for theoretical supports.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Thomas DR (2007) Loss of skeletal muscle mass in aging: examining the relationship of starvation, sarcopenia and cachexia. Clin Nutr 26(4):389–399. CrossRefPubMedGoogle Scholar
  2. 2.
    van Horssen R, Ten Hagen TL, Eggermont AM (2006) TNF-alpha in cancer treatment: molecular insights, antitumor effects, and clinical utility. Oncologist 11(4):397–408. CrossRefPubMedGoogle Scholar
  3. 3.
    Powers SK, Kavazis AN, McClung JM (2007) Oxidative stress and disuse muscle atrophy. J Appl Physiol 102(6):2389–2397. CrossRefPubMedGoogle Scholar
  4. 4.
    Stewart CE, Newcomb PV, Holly JM (2004) Multifaceted roles of TNF-alpha in myoblast destruction: a multitude of signal transduction pathways. J Cell Physiol 198(2):237–247. CrossRefPubMedGoogle Scholar
  5. 5.
    Chen D, Liu J, Lu L, Huang Y, Wang Y, Wang M, Liu Y, Xie D, Chen J, Diao J, Wei L, Wang M (2016) Emodin attenuates TNF-alpha-induced apoptosis and autophagy in mouse C2C12 myoblasts though the phosphorylation of Akt. Int Immunopharmacol 34:107–113. CrossRefPubMedGoogle Scholar
  6. 6.
    Huppertz B, Tews DS, Kaufmann P (2001) Apoptosis and syncytial fusion in human placental trophoblast and skeletal muscle. Int Rev Cytol 205:215–253CrossRefPubMedGoogle Scholar
  7. 7.
    Sandri M, Carraro U (1999) Apoptosis of skeletal muscles during development and disease. Int J Biochem Cell Biol 31(12):1373–1390CrossRefPubMedGoogle Scholar
  8. 8.
    Jejurikar SS, Henkelman EA, Cederna PS, Marcelo CL, Urbanchek MG, Kuzon WM Jr (2006) Aging increases the susceptibility of skeletal muscle derived satellite cells to apoptosis. Exp Gerontol 41(9):828–836. CrossRefPubMedGoogle Scholar
  9. 9.
    Deldicque L (2013) Endoplasmic reticulum stress in human skeletal muscle: any contribution to sarcopenia? Front Physiol 4:236. CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Deldicque L, Hespel P, Francaux M (2012) Endoplasmic reticulum stress in skeletal muscle: origin and metabolic consequences. Exerc Sport Sci Rev 40(1):43–49. CrossRefPubMedGoogle Scholar
  11. 11.
    Deldicque L, Van Proeyen K, Francaux M, Hespel P (2011) The unfolded protein response in human skeletal muscle is not involved in the onset of glucose tolerance impairment induced by a fat-rich diet. Eur J Appl Physiol 111(7):1553–1558. CrossRefPubMedGoogle Scholar
  12. 12.
    Finkel T, Holbrook NJ (2000) Oxidants, oxidative stress and the biology of ageing. Nature 408(6809):239–247. CrossRefPubMedGoogle Scholar
  13. 13.
    Ron D, Walter P (2007) Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol 8(7):519–529. CrossRefPubMedGoogle Scholar
  14. 14.
    Zhang K, Kaufman RJ (2008) Identification and characterization of endoplasmic reticulum stress-induced apoptosis in vivo. Methods Enzymol 442:395–419. CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Szegezdi E, Logue SE, Gorman AM, Samali A (2006) Mediators of endoplasmic reticulum stress-induced apoptosis. EMBO Rep 7(9):880–885. CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Fels DR, Koumenis C (2006) The PERK/eIF2alpha/ATF4 module of the UPR in hypoxia resistance and tumor growth. Cancer Biol Ther 5(7):723–728CrossRefPubMedGoogle Scholar
  17. 17.
    Lai E, Teodoro T, Volchuk A (2007) Endoplasmic reticulum stress: signaling the unfolded protein response. Physiology 22:193–201. CrossRefPubMedGoogle Scholar
  18. 18.
    Chiu FL, Lin JK (2008) Tomatidine inhibits iNOS and COX-2 through suppression of NF-kappaB and JNK pathways in LPS-stimulated mouse macrophages. FEBS Lett 582(16):2407–2412. CrossRefPubMedGoogle Scholar
  19. 19.
    Ebert SM, Dyle MC, Bullard SA, Dierdorff JM, Murry DJ, Fox DK, Bongers KS, Lira VA, Meyerholz DK, Talley JJ, Adams CM (2015) Identification and small molecule inhibition of an activating transcription factor 4 (ATF4)-dependent pathway to age-related skeletal muscle weakness and atrophy. J Biol Chem 290(42):25497–25511. CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Schiaffino S, Dyar KA, Ciciliot S, Blaauw B, Sandri M (2013) Mechanisms regulating skeletal muscle growth and atrophy. FEBS J 280(17):4294–4314. CrossRefPubMedGoogle Scholar
  21. 21.
    Kandarian SC, Jackman RW (2006) Intracellular signaling during skeletal muscle atrophy. Muscle Nerve 33(2):155–165. CrossRefPubMedGoogle Scholar
  22. 22.
    Tolosa L, Morla M, Iglesias A, Busquets X, Llado J, Olmos G (2005) IFN-gamma prevents TNF-alpha-induced apoptosis in C2C12 myotubes through down-regulation of TNF-R2 and increased NF-kappaB activity. Cell Signal 17(11):1333–1342. CrossRefPubMedGoogle Scholar
  23. 23.
    Gallo D, Gesmundo I, Trovato L, Pera G, Gargantini E, Minetto MA, Ghigo E, Granata R (2015) GH-releasing hormone promotes survival and prevents TNF-alpha-induced apoptosis and atrophy in C2C12 myotubes. Endocrinology 156(9):3239–3252. CrossRefPubMedGoogle Scholar
  24. 24.
    Baud V, Karin M (2001) Signal transduction by tumor necrosis factor and its relatives. Trends Cell Biol 11(9):372–377CrossRefPubMedGoogle Scholar
  25. 25.
    Dirks AJ, Leeuwenburgh C (2006) Tumor necrosis factor alpha signaling in skeletal muscle: effects of age and caloric restriction. J Nutr Biochem 17(8):501–508. CrossRefPubMedGoogle Scholar
  26. 26.
    Guo H, Chen L, Cui H, Peng X, Fang J, Zuo Z, Deng J, Wang X, Wu B (2015) Research advances on pathways of nickel-induced apoptosis. Int J Mol Sci 17(1).
  27. 27.
    Faitova J, Krekac D, Hrstka R, Vojtesek B (2006) Endoplasmic reticulum stress and apoptosis. Cell Mol Biol Lett 11(4):488–505. CrossRefPubMedGoogle Scholar
  28. 28.
    Garg AD, Kaczmarek A, Krysko O, Vandenabeele P, Krysko DV, Agostinis P (2012) ER stress-induced inflammation: does it aid or impede disease progression? Trends Mol Med 18(10):589–598. CrossRefPubMedGoogle Scholar
  29. 29.
    Zhang C, Kawauchi J, Adachi MT, Hashimoto Y, Oshiro S, Aso T, Kitajima S (2001) Activation of JNK and transcriptional repressor ATF3/LRF1 through the IRE1/TRAF2 pathway is implicated in human vascular endothelial cell death by homocysteine. Biochem Biophys Res Commun 289(3):718–724. CrossRefPubMedGoogle Scholar
  30. 30.
    da Silva DC, Andrade PB, Valentao P, Pereira DM (2017) Neurotoxicity of the steroidal alkaloids tomatine and tomatidine is RIP1 kinase- and caspase-independent and involves the eIF2alpha branch of the endoplasmic reticulum. J Steroid Biochem Mol Biol 171:178–186. CrossRefPubMedGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2017

Authors and Affiliations

  1. 1.Department of Physiology & Obesity-Mediated Disease Research CenterKeimyung University School of MedicineDaeguSouth Korea
  2. 2.Department of Immunology & Obesity-Mediated Disease Research CenterKeimyung University School of MedicineDaeguSouth Korea

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