Skip to main content
SpringerLink
Log in

The cardioprotective effect of postconditioning is mediated by ARC through inhibiting mitochondrial apoptotic pathway

  • Original Paper
  • Published:
Apoptosis Aims and scope Submit manuscript

Abstract

Objective

Postconditioning protects the heart against ischemia/reperfusion injury by inhibiting cardiomyocyte apoptosis. However, the molecular mechanism by which postconditioning suppresses apoptosis remains to be fully understood. Apoptosis repressor with caspase recruitment domain (ARC) has been demonstrated to possess the ability to protect cardiomyocytes from apoptosis induced by ischemia/reperfusion. It is not yet clear as to whether ARC contributes to the inhibitory effect of postconditioning against cardiomyocyte apoptosis.

Methods

The cultured cardiomyocytes from 1-day old male Sprague–Dawley rats were exposed to 3 h hypoxia followed by 3 h of reoxygenation. Cells were postconditioned by three cycles each of 5 min reoxygenation and 5 min hypoxia before 3 h of reoxygenation.

Results

Hypoxia/reoxygenation led to a decrease of endogenous ARC protein levels. In contrast, postconditioning could block the reduction of endogenous ARC protein levels. Interestingly, inhibition of endogenous ARC expression by ARC antisense oligodeoxynucleotides reduced the inhibitory effect of postconditioning against apoptosis. Furthermore, our data showed that postconditioning suppressed the loss of mitochondrial membrane potential, Bax activation and the release of mitochondrial cytochrome c to cytosol. However, these inhibitory effects of postconditioning disappeared upon knockdown of endogenous ARC.

Conclusion

Our data for the first time demonstrate that ARC plays an essential role in mediating the cardioprotective effect of postconditioning against apoptosis initiated by the mitochondrial pathway.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Braunwald E, Kloner RA (1985) Myocardial reperfusion: a double-edged sword? J Clin Invest 76:1713–1719. doi:10.1172/JCI112160

    Article  PubMed  CAS  Google Scholar 

  2. Diaz RJ, Wilson GJ (2004) Modifying the first minute of reperfusion: potential for myocardial salvage. Cardiovasc Res 62:4–6. doi:10.1016/j.cardiores.2004.01.036

    Article  PubMed  CAS  Google Scholar 

  3. Zhao ZQ, Corvera JS, Halkos ME et al (2003) Inhibition of myocardial injury by ischemic postconditioning during reperfusion: comparison with ischemic preconditioning. Am J Physiol Heart Circ Physiol 285:579–588

    Google Scholar 

  4. Kin H, Zhao ZQ, Sun HY et al (2004) Postconditioning attenuates myocardial ischemia–reperfusion injury by inhibiting events in the early minutes of reperfusion. Cardiovasc Res 62:74–85. doi:10.1016/j.cardiores.2004.01.006

    Article  PubMed  CAS  Google Scholar 

  5. Sun HY, Wang NP, Kerendi F et al (2005) Hypoxic postconditioning reduces cardiomyocyte loss by inhibiting ROS generation and intracellular Ca2+ overload. Am J Physiol Heart Circ Physiol 288:1900–1908. doi:10.1152/ajpheart.01244.2003

    Article  Google Scholar 

  6. Sun HY, Wang NP, Halkos M et al (2006) Postconditioning attenuates cardiomyocyte apoptosis via inhibition of JNK and p38 mitogen-activated protein kinase signaling pathways. Apoptosis 11:1583–1593. doi:10.1007/s10495-006-9037-8

    Article  PubMed  CAS  Google Scholar 

  7. Wang HC, Zhang HF, Guo WY et al (2006) Hypoxic postconditioning enhances the survival and inhibits apoptosis of cardiomyocytes following reoxygenation: role of peroxynitrite formation. Apoptosis 11:1453–1460. doi:10.1007/s10495-006-7786-z

    Article  PubMed  CAS  Google Scholar 

  8. Weihrauch D, Krolikowski JG, Bienengraeber M et al (2005) Morphine enhances isoflurane-induced postconditioning against myocardial infarction: the role of phosphatidylinositol-3-kinase and opioid receptors in rabbits. Anesth Analg 101(4):942–949. doi:10.1213/01.ane.0000171931.08371.a2

    Article  PubMed  CAS  Google Scholar 

  9. Koseki T, Inohara N, Chen S et al (1998) ARC, an inhibitor of apoptosis expressed in skeletal muscle and heart that interacts selectively with caspases. Proc Natl Acad Sci USA 95:5156–5160. doi:10.1073/pnas.95.9.5156

    Article  PubMed  CAS  Google Scholar 

  10. Nam YJ, Mani K, Ashton AW, Peng CF, Krishnamurthy B, Hayakawa Y, Lee P, Korsmeyer SJ, Kitsis RN (2004) Inhibition of both the extrinsic and intrinsic death pathways through nonhomotypic death-fold interactions. Mol Cell 15:901–912. doi:10.1016/j.molcel.2004.08.020

    Article  PubMed  CAS  Google Scholar 

  11. Ekhterae D, Lin Z, Lundberg MS et al (1999) ARC inhibits cytochrome c release from mitochondria and protects against hypoxia-induced apoptosis in heart-derived H9c2 cells. Circ Res 85:70–77

    Google Scholar 

  12. Gustafsson AB, Sayen MR, Williams SD et al (2002) TAT protein transduction into isolated perfused hearts, TAT-apoptosis repressor with caspase recruitment domain is cardioprotective. Circulation 106:735–739

    PubMed  CAS  Google Scholar 

  13. Jo DG, Jun JI, Chang JW, Hong YM, Song S, Cho DH, Shim SM, Lee HJ, Cho C, Kim do K, Jung YK (2004) Calcium binding of ARC mediates regulation of caspase 8 and cell death. Mol Cell Biol 24:9763–9770. doi:10.1128/MCB.24.22.9763-9770.2004

    Article  PubMed  CAS  Google Scholar 

  14. Pyol JO, Nah J, Kim HJ et al (2008) Protection of cardiomyocytes from ischemia/hypoxic cel death via Drbp1 and pMe2GlyDH in cardio-specific ARC transgenic mice. J Biol Chem 283:30707–30714

    Article  Google Scholar 

  15. Donath S, Li P, Willenbockel C et al (2006) Apoptosis repressor with caspase recruitment domain is required for cardioprotection in response to biomechanical and ischemic stress. Circulation 113:1203–1212. doi:10.1161/CIRCULATIONAHA.105.576785

    Article  PubMed  CAS  Google Scholar 

  16. Argaud L, Gateau-Roesch O, Raisky O et al (2005) Postconditioning inhibits mitochondrial permeability transition. Circulation 111:194–197. doi:10.1161/01.CIR.0000151290.04952.3B

    Article  PubMed  CAS  Google Scholar 

  17. Neuss M, Monticone R, Lundberg MS et al (2001) The apoptotic regulatory protein ARC (apoptosis repressor with caspase recruitment domain) prevents oxidant stress-mediated cell death by preserving mitochondrial function. J Biol Chem 276:33915–33922. doi:10.1074/jbc.M104080200

    Article  PubMed  CAS  Google Scholar 

  18. Li YZ, Liu XH, Zhu XM et al (2007) ARC contributes to the inhibitory effect of preconditioning on cardiomyocyte apoptosis. Apoptosis 12(9):1589–1595. doi:10.1007/s10495-007-0094-4

    Article  PubMed  CAS  Google Scholar 

  19. Salvioli S, Ardizzoni A, Franceschi C et al (1997) JC-1, but not DiOC6(3) or rhodamine 123, is a reliable fluorescent probe to assess delta psi changes in intact cells: implications for studies on mitochondrial functionality during apoptosis. FEBS Lett 411:77–82. doi:10.1016/S0014-5793(97)00669-8

    Article  PubMed  CAS  Google Scholar 

  20. Desagher S, Osen-Sand A, Nichols A et al (1999) Bid-induced conformational change of Bax is responsible for mitochondrial cytochrome c release during apoptosis. J Cell Biol 144(5):891–901. doi:10.1083/jcb.144.5.891

    Article  PubMed  CAS  Google Scholar 

  21. Gustafsson AB, Tsai JG, Logue SE et al (2004) Apoptosis repressor with caspase recruitment domain protects against cell death by interfering with Bax activation. J Biol Chem 279(20):21233–21238. doi:10.1074/jbc.M400695200

    Article  PubMed  CAS  Google Scholar 

  22. Tsang A, Hausenloy DJ, Mocanu MM et al (2004) Postconditioning: a form of “modified reperfusion” protects the myocardium by activating the phosphatidylinositol 3-kinase-Akt pathway. Circ Res 95:230–232. doi:10.1161/01.RES.0000138303.76488.fe

    Article  PubMed  CAS  Google Scholar 

  23. Uchiyama T, Engelman RM, Maulik N et al (2004) Role of Akt signaling in mitochondrial survival pathway triggered by hypoxic preconditioning. Circulation 109:3042–3049. doi:10.1161/01.CIR.0000130647.29030.90

    Article  PubMed  CAS  Google Scholar 

  24. Csonka C, Csont T, Onody A et al (2001) Preconditioning decreases ischemia/reperfusion-induced peroxynitrite formation. Biochem Biophys Res Commun 285(5):1217–1219. doi:10.1006/bbrc.2001.5308

    Article  PubMed  CAS  Google Scholar 

  25. Vinten-Johansen J, Zhao ZQ, Zatta AJ et al (2005) Postconditioning: a new link in nature’s armor against myocardial ischemia–reperfusion injury. Basic Res Cardiol 100:295–310. doi:10.1007/s00395-005-0523-x

    Article  PubMed  CAS  Google Scholar 

  26. Yang XM, Proctor JB, Cui L et al (2004) Multiple, brief coronary occlusions during early reperfusion protect rabbit hearts by targeting cell signaling pathways. J Am Coll Cardiol 44:1103–1110. doi:10.1016/j.jacc.2004.05.060

    Article  PubMed  Google Scholar 

  27. Kim SO, Baines CP, Critz SD et al (1999) Ischemia induced activation of heat shock protein 27 kinases and casein kinase 2 in the preconditioned rabbit heart. Biochem Cell Biol Biochim Biol Cell 77:559–567. doi:10.1139/bcb-77-6-559

    Article  CAS  Google Scholar 

  28. Huang DC, Strasser A (2000) BH3-only proteins-essential initiators of apoptotic cell death. Cell 103:839–842. doi:10.1016/S0092-8674(00)00187-2

    Article  PubMed  CAS  Google Scholar 

  29. Nam YJ, Mani K, Ashton AW et al (2004) Inhibition of both the extrinsic and intrinsic death pathways through nonhomotypic death-fold interactions. Mol Cell 15(6):901–912. doi:10.1016/j.molcel.2004.08.020

    Article  PubMed  CAS  Google Scholar 

  30. Li YZ, Lu DY, Tan WQ et al (2008) p53 initiates apoptosis by transcriptionally targeting the antiapoptotic protein ARC. Mol Cell Biol 28(2):564–574. doi:10.1128/MCB.00738-07

    Article  PubMed  CAS  Google Scholar 

  31. Nam YJ, Mani K, Wu L et al (2007) The apoptosis inhibitor ARC undergoes ubiquitin-proteasomal-mediated degradation in response to death stimuli. J Biol Chem 282:5522–5528. doi:10.1074/jbc.M609186200

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from Chinese National Natural Scientific Fund (Li30570735, Li30772297); National Basic Research Program of China (Liu2007CB512003).

Author information

Authors and Affiliations

  1. Department of Pathophysiology, Institute of Basic Medical Science, Chinese PLA General Hospital, 100853, Beijing, China

    YuZhen Li, Xinlan Ge & XiuHua Liu

Authors
  1. YuZhen Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xinlan Ge
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. XiuHua Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to YuZhen Li.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, Y., Ge, X. & Liu, X. The cardioprotective effect of postconditioning is mediated by ARC through inhibiting mitochondrial apoptotic pathway. Apoptosis 14, 164–172 (2009). https://doi.org/10.1007/s10495-008-0296-4

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10495-008-0296-4

Keywords

Search

Navigation