Neurological Sciences

, Volume 35, Issue 11, pp 1691–1699 | Cite as

Mild hypothermia alleviates excessive autophagy and mitophagy in a rat model of asphyxial cardiac arrest

  • Jian Lu
  • Hui-Yin Qian
  • Li-Jun LiuEmail author
  • Bao-Chun Zhou
  • Yan Xiao
  • Jin-Ning Mao
  • Guo-Yin An
  • Ming-Zhong Rui
  • Tao Wang
  • Chang-Lai Zhu
Original Article


Mild hypothermia is an effective therapeutic strategy to improve poor neurological outcomes in patients following cardiac arrest (CA). However, the underlying mechanism remains unclear. The aim of the study was to evaluate the effect of mild hypothermia on intracellular autophagy and mitophagy in hippocampal neurons in a rat model of CA. CA was induced in Sprague–Dawley (SD) rats by asphyxia for 5 min. After successful resuscitation, the surviving rats were randomly divided into two groups, the normothermia (NT) group and the hypothermia (HT) group. Mild hypothermia (32 °C) was induced following CA for 4 h, and animals were rewarmed at a rate of 0.5 °C/h. Neurologic deficit scores (NDS) were used to determine the status of neurological function. Cytoplasmic and mitochondrial protein from the hippocampus was extracted, and the expression of LC3B-II/I and Parkin were measured as markers of intracellular autophagy and mitophagy, respectively. Of the 60 rats that underwent CA, 44 were successfully resuscitated (73 %), and 33 survived until the end of the experiment (55 %). Mild hypothermia maintained eumorphism of nuclear and mitochondrial structures and significantly improved NDS (p < 0.05). Expression of LC3B-II/I and Parkin in hippocampal nerve cells were significantly increased (p < 0.05) in the NT group relative to the control. Meanwhile, mild hypothermia reduced the level of LC3B-II/I and Parkin (p < 0.05) relative to the NT group. Mild hypothermia protected mitochondria and improved neurological function following CA and resuscitation after ischemia/reperfusion (I/R) injury, likely by reducing excessive autophagy and mitophagy in neurons.


Mild hypothermia Autophagy Mitophagy Cardiac arrest Ischemic/reperfusion injury 



The authors would like to thank Prof Zi-Tong Huang and Prof Xiang-Shao Fang of the Affiliated Sun Yat-sen Memory Hospital of Zhongshan University and Director Ai-Dong Wang of the Experimental Center of the Second Affiliated Hospital of Soochow University, the Experimental Center of Suzhou Health College for their technical assistance.

Conflict of interest

The authors declare they have no existing conflict of interest.


  1. 1.
    Neumar RW, Nolan JP, Adrie C, Aibiki M, Berg RA, Böttiger BW, Callaway C, Clark RS, Geocadin RG, Jauch EC (2008) Post-cardiac arrest syndrome epidemiology, pathophysiology, treatment, and prognostication. A consensus statement from the International Liaison Committee On Resuscitation (American Heart Association, Australian and New Zealand Council on Resuscitation, European Resuscitation Council, Heart and Stroke Foundation of Canada, InterAmerican Heart Foundation, Resuscitation Council of Asia, and the Resuscitation Council of Southern Africa); the American Heart Association Emergency Cardiovascular Care Committee; the Council on Cardiovascular Surgery and Anesthesia; the Council on Cardiopulmonary, Perioperative, and Critical Care; the Council on Clinical Cardiology; and the Stroke Council. Circulation 118(23):2452–2483PubMedCrossRefGoogle Scholar
  2. 2.
    Shin TG, Jo IJ, Song HG, Sim MS, Song KJ (2012) Improving survival rate of patients with in-hospital cardiac arrest: five years of experience in a single center in Korea. J Korean Med Sci 27(2):146–152PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Ramachandran SK, Mhyre J, Kheterpal S, Christensen RE, Tallman K, Morris M, Chan PS (2013) Predictors of survival from perioperative cardiopulmonary arrests: a retrospective analysis of 2,524 events from the Get With The Guidelines-Resuscitation Registry. Anesthesiology 119(6):1322–1339PubMedCrossRefGoogle Scholar
  4. 4.
    Polderman KH (2008) Induced hypothermia and fever control for prevention and treatment of neurological injuries. Lancet 371(9628):1955–1969PubMedCrossRefGoogle Scholar
  5. 5.
    Busl KM, Greer DM (2010) Hypoxic-ischemic brain injury: pathophysiology, neuropathology and mechanisms. NeuroRehabilitation 26(1):5–13PubMedGoogle Scholar
  6. 6.
    Nikolov NM, Cunningham AJ (2003) Mild therapeutic hypothermia to improve the neurologic outcome after cardiac arrest. N Engl J Med 47(4):219–220Google Scholar
  7. 7.
    Bernard SA, Gray TW, Buist MD, Jones BM, Silvester W, Gutteridge G, Smith K (2002) Treatment of comatose survivors of out-of-hospital cardiac arrest with induced hypothermia. N Engl J Med 346(8):557–563PubMedCrossRefGoogle Scholar
  8. 8.
    Marion D, Bullock MR (2009) Current and future role of therapeutic hypothermia. J Neurotrauma 26(3):455–467PubMedCrossRefGoogle Scholar
  9. 9.
    Gong P, Li C-S, Hua R, Zhao H, Tang Z-R, Mei X, Zhang M-Y, Cui J (2012) Mild hypothermia attenuates mitochondrial oxidative stress by protecting respiratory enzymes and upregulating MnSOD in a pig model of cardiac arrest. PLoS One 7(4):e35313PubMedCentralPubMedCrossRefGoogle Scholar
  10. 10.
    Zhang X, Yan H, Yuan Y, Gao J, Shen Z, Cheng Y, Shen Y, Wang RR, Wang X, Hu WW, Wang G, Chen Z (2013) Cerebral ischemia-reperfusion-induced autophagy protects against neuronal injury by mitochondrial clearance. Autophagy 9(9):1321–1333PubMedCrossRefGoogle Scholar
  11. 11.
    Wang K, Klionsky DJ (2011) Mitochondria removal by autophagy. Autophagy 7(3):297–300PubMedCentralPubMedCrossRefGoogle Scholar
  12. 12.
    Geocadin R, Ghodadra R, Kimura T, Lei H, Sherman D, Hanley D, Thakor N (2000) A novel quantitative EEG injury measure of global cerebral ischemia. Clin Neurophysiol 111(10):1779–1787PubMedCrossRefGoogle Scholar
  13. 13.
    Chan PH (2005) Mitochondrial dysfunction and oxidative stress as determinants of cell death/survival in stroke. Ann NY Acad Sci 1042(1):203–209PubMedCrossRefGoogle Scholar
  14. 14.
    Szabó C (2003) Multiple pathways of peroxynitrite cytotoxicity. Toxicol Lett 140:105–112PubMedCrossRefGoogle Scholar
  15. 15.
    Stowe DF, Camara AK (2009) Mitochondrial reactive oxygen species production in excitable cells: modulators of mitochondrial and cell function. Antioxid Redox Signal 11(6):1373–1414PubMedCentralPubMedCrossRefGoogle Scholar
  16. 16.
    Klionsky DJ, Abdalla FC, Abeliovich H, Abraham RT, Acevedo-Arozena A, Adeli K, Agholme L, Agnello M, Agostinis P, Aguirre-Ghiso JA (2012) Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy 8(4):445–544PubMedCentralPubMedCrossRefGoogle Scholar
  17. 17.
    Ni H, Gong Y, Yan J-z, Zhang L-l (2010) Autophagy inhibitor 3-methyladenine regulates the expression of LC3, Beclin-1 and ZnTs in rat cerebral cortex following recurrent neonatal seizures. World 1(3):216–223Google Scholar
  18. 18.
    Adhami F, Liao G, Morozov YM, Schloemer A, Schmithorst VJ, Lorenz JN, Dunn RS, Vorhees CV, Wills-Karp M, Degen JL (2006) Cerebral ischemia-hypoxia induces intravascular coagulation and autophagy. Am J Pathol 169(2):566–583PubMedCentralPubMedCrossRefGoogle Scholar
  19. 19.
    Koike M, Shibata M, Tadakoshi M, Gotoh K, Komatsu M, Waguri S, Kawahara N, Kuida K, Nagata S, Kominami E (2008) Inhibition of autophagy prevents hippocampal pyramidal neuron death after hypoxic-ischemic injury. Am J Pathol 172(2):454–469PubMedCentralPubMedCrossRefGoogle Scholar
  20. 20.
    Starkov Anatoly A (2008) The role of mitochondria in reactive oxygen species metabolism and signaling. Ann N Y Acad Sci 1147:37–52PubMedCentralPubMedCrossRefGoogle Scholar
  21. 21.
    Huang Q, Shen HM (2009) To die or to live: the dual role of poly(ADP-ribose) polymerase-1 in autophagy and necrosis under oxidative stress and DNA damage. Autophagy 5(2):273–276PubMedCrossRefGoogle Scholar
  22. 22.
    Geisler S, Holmström KM, Treis A, Skujat D, Weber SS, Fiesel FC, Kahle PJ, Springer W (2010) The PINK1/Parkin-mediated mitophagy is compromised by PD-associated mutations. Autophagy 6(7):871–878PubMedCrossRefGoogle Scholar
  23. 23.
    Zhao H, Steinberg GK, Sapolsky RM (2007) General versus specific actions of mild-moderate hypothermia in attenuating cerebral ischemic damage. J Cereb Blood Flow Metab 27(12):1879–1894PubMedCrossRefGoogle Scholar
  24. 24.
    Zhao H, Yenari MA, Cheng D, Sapolsky RM, Steinberg GK (2005) Biphasic cytochrome c release after transient global ischemia and its inhibition by hypothermia. J Cereb Blood Flow Metab 25(9):1119–1129PubMedCrossRefGoogle Scholar
  25. 25.
    Xu L, Yenari MA, Steinberg GK, Giffard RG (2002) Mild hypothermia reduces apoptosis of mouse neurons in vitro early in the cascade. J Cereb Blood Flow Metab 22(1):21–28PubMedCrossRefGoogle Scholar
  26. 26.
    Meybohm P, Gruenewald M, Albrecht M, Zacharowski KD, Lucius R, Zitta K, Koch A, Tran N, Scholz J, Bein B (2009) Hypothermia and postconditioning after cardiopulmonary resuscitation reduce cardiac dysfunction by modulating inflammation, apoptosis and remodeling. PLoS One 4(10):e7588PubMedCentralPubMedCrossRefGoogle Scholar
  27. 27.
    Shen J, Chen C, Xu J, Fu G (2013) Reduction of autophagy: another potential mechanism of cardioprotective effect of mild hypothermia? Int J Cardiol 168(5):4810–4811PubMedCrossRefGoogle Scholar
  28. 28.
    Cheng B-C, Huang H-S, Chao C-M, Hsu C–C, Chen C-Y, Chang C-P (2013) Hypothermia may attenuate ischemia/reperfusion-induced cardiomyocyte death by reducing autophagy. Int J Cardiol 168(3):2064–2069PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Italia 2014

Authors and Affiliations

  • Jian Lu
    • 1
  • Hui-Yin Qian
    • 1
  • Li-Jun Liu
    • 1
    Email author
  • Bao-Chun Zhou
    • 1
  • Yan Xiao
    • 1
  • Jin-Ning Mao
    • 2
  • Guo-Yin An
    • 2
  • Ming-Zhong Rui
    • 3
  • Tao Wang
    • 4
  • Chang-Lai Zhu
    • 5
  1. 1.Department of Emergency and Critical Care MedicineThe Second Affiliated Hospital of Soochow UniversitySuzhouPeople’s Republic of China
  2. 2.Department of CardiologyThe Second Affiliated Hospital of Soochow UniversitySuzhouPeople’s Republic of China
  3. 3.Department of HematologyThe Second Affiliated Hospital of Soochow UniversitySuzhouPeople’s Republic of China
  4. 4.Department of LaboratoryThe Second Affiliated Hospital of Soochow UniversitySuzhouPeople’s Republic of China
  5. 5.Electron Microscope RoomThe Affiliated Hospital of Nantong UniversityNantongPeople’s Republic of China

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