Cell Biology and Toxicology

, Volume 28, Issue 1, pp 19–29 | Cite as

Mitigation of H2O2-induced autophagic cell death by propofol in H9c2 cardiomyocytes

  • Ji Hye Ha
  • Hae Sook Noh
  • Il Woo Shin
  • Jong Ryeal Hahm
  • Deok Ryong Kim


Autophagy, a self-eating process, is responsible for degradation of long-lived proteins and damaged cellular proteins/organelles. Double-membrane autophagosomes, formed during the process, engulf proteins/organelles and fuse with lysosomes to degrade the contents. It is important to maintain cell homeostasis and many physiological processes including cellular responses to oxidative stress. Oxidative stress induced by myocardial infarction is a major factor of heart failures. In this study, we examined how propofol modulates hydrogen peroxide (H2O2)-induced autophagic cell death in H9c2 cardiomyocytes. H2O2 dramatically induced cell death, which was similarly reduced in the presence of either propofol or autophagy inhibitors (e.g., wortmannin), suggesting that propofol has a protective effect in H2O2-induced autophagic cell death. Acidic autophagic vacuoles were elevated in H2O2-treated H9c2 cells, but they were largely decreased in the presence of propofol. Furthermore, many autophagy-related proteins such as LC3-II, ATG proteins, p62, AMPK, and JNK were activated in H2O2-treated H9c2 cells and were significantly deactivated in the presence of propofol. These results show that propofol regulates oxidative stress-induced autophagic cell death in cardiomyocytes. We further suggest that propofol can act as a cardioprotectant in heart diseases.


AMPK Autophagic cell death Cardiomyocytes Hydrogen peroxide JNK Oxidative stress Propofol 



5′ AMP-activated protein kinase


Acridine orange


Autophagy-related gene




Janus kinase


Mitogen-activated protein kinase




Mammalian target of rapamycin


Poly (ADP-ribose) polymerase



This work was supported by a grant from the Korea Health Care Technology R&D Project, Ministry of Health and Welfare, Republic of Korea (A080403) and a National Research Foundation of Korea grant funded by the Korean government (NRF-2009-0071600 and R13-2005-012-02002-0).


  1. Arnaoutoglou H, Vretzakis G, Souliotis D, Cambili M, Galaris D, Papadopoulos G. The effects of propofol or sevoflurane on free radical production after tourniquet induced ischaemia-reperfusion injury during knee arthroplasty. Acta Anaesthesiol Belg. 2007;58:3–6.PubMedGoogle Scholar
  2. Black SC. In vivo models of myocardial ischemia and reperfusion injury: application to drug discovery and evaluation. J Pharmacol Toxicol Meth. 2000;43:153–67.CrossRefGoogle Scholar
  3. Chen Y, McMillan-Ward E, Kong J, Israels SJ, Gibson SB. Oxidative stress induces autophagic cell death independent of apoptosis in transformed and cancer cells. Cell Death Differ. 2008;15:171–82.PubMedCrossRefGoogle Scholar
  4. Choi SL, Kim SJ, Lee KT, Kim J, Mu J, Birnbaum MJ, et al. The regulation of AMP-activated protein kinase by H(2)O(2). Biochem Biophys Res Comm. 2001;287:92–7.PubMedCrossRefGoogle Scholar
  5. Cory S, Adams JM. The Bcl2 family: regulators of the cellular life-or-death switch. Nat Rev Canc. 2002;2:647–56.CrossRefGoogle Scholar
  6. Dadakhujaev S, Noh HS, Jung EJ, Hah YS, Kim CJ, Kim DR. The reduced catalase expression in TrkA-induced cells leads to autophagic cell death via ROS accumulation. Exp Cell Res. 2008;314:3094–106.CrossRefGoogle Scholar
  7. Hein S, Arnon E, Kostin S, Schonburg M, Elsasser A, Polyakova V, et al. Progression from compensated hypertrophy to failure in the pressure-overloaded human heart: structural deterioration and compensatory mechanisms. Circulation. 2003;107:984–91.PubMedCrossRefGoogle Scholar
  8. Hwang J, Lee S, Lee JT, Kwon TK, Kim DR, Kim H, et al. Gangliosides induce autophagic cell death in astrocytes. Br J Pharmacol. 2010;159:586–603.PubMedCrossRefGoogle Scholar
  9. Ichimura Y, Kominami E, Tanaka K, Komatsu M. Selective turnover of p62/A170/SQSTM1 by autophagy. Autophagy. 2008;4:1063–6.PubMedGoogle Scholar
  10. Jin YC, Kim W, Ha YM, Shin IW, Sohn JT, Kim HJ, et al. Propofol limits rat myocardial ischemia and reperfusion injury with an associated reduction in apoptotic cell death in vivo. Vasc Pharmacol. 2009;50:71–7.CrossRefGoogle Scholar
  11. Kamiya T, Hara H, Yamada H, Imai H, Inagaki N, Adachi T. Cobalt chloride decreases EC-SOD expression through intracellular ROS generation and p38-MAPK pathways in COS7 cells. Free Radic Res. 2008;42:949–56.PubMedCrossRefGoogle Scholar
  12. Kiffin R, Bandyopadhyay U, Cuervo AM. Oxidative stress and autophagy. Antioxidants and Redox Signaling. 2006;8:152–62.PubMedCrossRefGoogle Scholar
  13. Kimes BW, Brandt BL. Properties of a clonal muscle cell line from rat heart. Exp Cell Res. 1976;98:367–81.PubMedCrossRefGoogle Scholar
  14. Konishi H, Fujiyoshi T, Fykui Y, Matsuzaki H, Yamamoto T, Ono Y, et al. Activation of protein kinase B indeced by H2O2 and heat shock through distinct mechanisms dependent and independent of phosphpatidylinositol 3-kinase. J Biochem (Tokyo). 1999;126:1136–43.Google Scholar
  15. Kroemer G, Marino G, Levine B. Autophagy and the intergrated stress response. Mol cell. 2010;40:280–93.PubMedCrossRefGoogle Scholar
  16. Kubota C, Torii S, Hou N, Saito N, Yoshimoto Y, Imai H, et al. Constitutive reactive oxygen species generation from autophagosome/lysosome in neuronal oxidative toxicity. J Biol Chem. 2010;285:667–74.PubMedCrossRefGoogle Scholar
  17. Lang XH, Kleeman LK, Jiang HH, Gordon G, Goldman JE, Berry G, et al. Protection against fatal Sindbis virus encephalitis by Beclin, a novel Bcl-2-interaction protein. J Virol. 1998;72:8586–96.Google Scholar
  18. Levine B, Yuan J. Autophagy in cell death: an innocent convict? J Clin Invest. 2005;115:2679–88.PubMedCrossRefGoogle Scholar
  19. Mathew R, Karp CM, Beaudoin B, Vuong N, Chen G, Chen HY, et al. Autophagy suppresses tumorigenesis through elimination of p62. Cell. 2009;137:1062–75.PubMedCrossRefGoogle Scholar
  20. Matsui Y, Nakano N, Shao D, Gao S, Luo W, Hong C, et al. Lats2 is a negative regulator of myocyte size in the heart. Circ Res. 2008;103:1309–18.PubMedCrossRefGoogle Scholar
  21. Mizushima N, Levin B, Cuervo AM, Klionsky DJ. Autophagy fights disease through cellular self-digestion. Nature. 2008;451:1069–75.PubMedCrossRefGoogle Scholar
  22. Mizushima N, Yoshimori T, Levine B. Methods in mammalian autophagy research. Cell. 2010;140:313–26.PubMedCrossRefGoogle Scholar
  23. Noh HS, Shin IW, Ha JH, Hah YS, Baek SM, Kim DR. Propofol protects the autophagic cell death induced by the ischemia/reperfusion injury in rats. Mol Cells. 2010;30:455–60.PubMedCrossRefGoogle Scholar
  24. Puissant A, Robert G, Fenouille N, Luciano F, Cassuto JP, Raynaud S, et al. Resveratrol promotes autophagic cell death in chronic myelogenous leukemia cells via JNK-mediated p62/SQSTM1 expression and AMPK activation. Canc Res. 2010;70:1042–52.CrossRefGoogle Scholar
  25. Ryter SW, Kim HP, Hoetzel A, Park JW, Nakahira K, Wang X, et al. Mechanisms of cell death in oxidative stress. Antioxidants and Redox Signaling. 2007;9:49–89.PubMedCrossRefGoogle Scholar
  26. Semenza GL, Agani F, Feldser D, Iyer N, Kotch L, Laughner E, et al. Hypoxia, HIF-1, and the pathophysiology of common human diseases. Adv Exp Med Biol. 2000;475:123–30.PubMedCrossRefGoogle Scholar
  27. Shimizu S, Konishi A, Nishida Y, Mizuta T, Nishina H, Yamamoto A, et al. Involvement of JNK in the regulation of autophagic cell death. Oncogene. 2010;29:2070–82.PubMedCrossRefGoogle Scholar
  28. Shintani T, Klionsky DJ. Autophagy in health and disease: a double-edged sword. Science. 2004;306:990–5.PubMedCrossRefGoogle Scholar
  29. Walsh SK, Hepburn CY, Kane KA, Wainwright CL. Acute administration of cannabidiol in vivo suppresses ischaemia-induced cardiac arrhythmias and reduces infarct size when given at reperfusion. Br J Pharmacol. 2010;160:1234–42.PubMedCrossRefGoogle Scholar
  30. Wang X, McCullough KD, Franke TF, Holbrook NJ. Epidermal growth factor receptor-dependent Akt activation by oxidative stress enhances cell survival. J Biol Chem. 2000;275:14624–31.PubMedCrossRefGoogle Scholar
  31. Wang H, Xue Z, Wang Q, Feng X, Shen Z. Propofol protects hepatic L02 cells from hydrogen peroxide-induced apoptosis via activation of extracellular signal-regulated kinases pathway. Anesth Analg. 2008;107:534–40.PubMedCrossRefGoogle Scholar
  32. Wang B, Shravah J, Luo H, Raedschelders K, Chen DDY, Ansley DM. Propofol protects against hydrogen peroxide-induced injury in cardiac H9c2 cells via Akt activation and Bcl-2 up-regulation. Biochem Biophy Res Comm. 2009;389:105–11.CrossRefGoogle Scholar
  33. Wu X, Zheng Y, Cui Y, Zhu L, Lu Y, Chen H. Propofol attenuates oxidative stress-induced PC12 cell injury via p38 MAP kinase dependent pathway. Acta Pharmacol Sin. 2007;28:1123–8.PubMedCrossRefGoogle Scholar
  34. Yang J, Klionsky DJ. Eaten alive: a history of macroautophagy. Nature Cell Biol. 2010;12:814–22.PubMedCrossRefGoogle Scholar
  35. Zalckvar E, Berissi H, Eisenstein M, Kimchi A. Phosphorylation of Beclin 1 by DAP-kinase promotes autophagy by weakening its interactions with Bcl-2 and Bcl-XL. Autophagy. 2009;5:720–2.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Ji Hye Ha
    • 1
    • 4
  • Hae Sook Noh
    • 1
    • 4
  • Il Woo Shin
    • 2
    • 4
  • Jong Ryeal Hahm
    • 3
    • 4
  • Deok Ryong Kim
    • 1
    • 4
  1. 1.Department of BiochemistryGyeongsang National University School of MedicineJinjuRepublic of Korea
  2. 2.Department Anesthesiology and Pain MedicineGyeongsang National University School of MedicineJinjuRepublic of Korea
  3. 3.Department Internal MedicineGyeongsang National University School of MedicineJinjuRepublic of Korea
  4. 4.Institute of Health SciencesGyeongsang National University School of MedicineJinjuRepublic of Korea

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