Molecular and Cellular Biochemistry

, Volume 370, Issue 1–2, pp 163–171 | Cite as

Calcineurin B subunit acts as a potential agent for preventing cardiac ischemia/reperfusion injury

  • Junxia Guo
  • Shengquan Mi
  • Jing Li
  • Wei Liu
  • Yanxia Yin
  • Qun Wei


Calcineurin B subunit (CnB) is the regulatory subunit of calcineurin (Cn), a Ca2+/calmodulin-dependent serine/threonine protein phosphatase. It has been reported that mice deleting the CnB gene lose nearly all Cn activity and show poor tolerance to cardiac stress; CnB gene expression is downregulated in the hearts of rats that have suffered ischemia/reperfusion (I/R) injury. Therefore, we wonder whether injection of exogenous CnB protein can prevent the rats from suffering I/R injury. In cardiomyocytes, fluorogenic labeling shows that exogenous CnB quickly enters the cell. Pretreatment of cardiomyocytes with CnB reduces apoptosis in response to hypoxia/reoxygenation injury (an in vitro model mimicking ischemia/reperfusion injury), and CsA reverses this effect by inhibiting Cn activity. Furthermore, CnB upregulates Bcl-2 and Bcl-XL expression in the process of hypoxia/reoxygenation injury, which may contribute to protecting cardiomyocytes against apoptosis. In vivo experiments shows that pretreatment with CnB improves cardiac contractile function and reduces the frequency of arrhythmias induced by global I/R injury. These findings reveal a novel function for CnB protein in cardiac stress response and suggest a possible application of CnB in coronary disease therapy.


Calcineurin CnB Cardiomyocyte Ischemia/reperfusion injury 



This work was partially supported by the National Natural Science Foundation of China, the International Cooperation Project, the National Important Novel Medicine Research Project, and the Fundamental Research Funds for the Central Universities.


  1. 1.
    Machado NG, Alves MG, Carvalho RA, Oliveira PJ (2009) Mitochondrial involvement in cardiac apoptosis during ischemia and reperfusion: can we close the box? Cardiovasc Toxicol 9:211–227PubMedCrossRefGoogle Scholar
  2. 2.
    Ferdinandy P, Schulz R, Baxter GF (2007) Interaction of cardiovascular risk factors with myocardial ischemia/reperfusion injury, preconditioning, and postconditioning. Pharmacol Rev 59:418–458PubMedCrossRefGoogle Scholar
  3. 3.
    Maulik N, Yoshida T, Engelman RM, Deaton D, Flack JE (1998) Ischemic preconditioning attenuates apoptotic cell death associated with ischemia/reperfusion. Mol Cell Biochem 186:139–145PubMedCrossRefGoogle Scholar
  4. 4.
    Bansal P, Gupta SK, Ojha SK, Nandave M, Mittal R, Kumari S, Arya DS (2006) Cardioprotective effect of lycopene in the experimental model of myocardial ischemia–reperfusion injury. Mol Cell Biochem 289:1–9PubMedCrossRefGoogle Scholar
  5. 5.
    Rietz B, Isensee H, Strobach H, Makdessi S, Jacob R (1993) Cardioprotective actions of wild garlic (Allium ursinum) in ischemia and reperfusion. Mol Cell Biochem 119:143–150PubMedCrossRefGoogle Scholar
  6. 6.
    Kloner RA, Rezkalla SH (2006) Preconditioning, postconditioning and their application to clinical cardiology. Cardiovasc Res 70:297–307PubMedCrossRefGoogle Scholar
  7. 7.
    Kurji K, Sharma RK (2010) Potential role of calcineurin in pathogenic conditions. Mol Cell Biochem 338:133–141PubMedCrossRefGoogle Scholar
  8. 8.
    Wu H, Peisley A, Graef IA, Crabtree GR (2007) NFAT signaling and the invention of vertebrates. Trends Cell Biol 17:251–260PubMedCrossRefGoogle Scholar
  9. 9.
    Hogan PG, Chen L, Nardone J, Rao A (2003) Transcriptional regulation by calcium, calcineurin, and NFAT. Genes Dev 17:2205–2232PubMedCrossRefGoogle Scholar
  10. 10.
    Molkentin JD, Lu JR, Antos CL, Markham B, Richardson J, Robbins J, Grant SR, Olson EN (1998) A calcineurin-dependent transcriptional pathway for cardiac hypertrophy. Cell 93:215–228PubMedCrossRefGoogle Scholar
  11. 11.
    Vega RB, Bassel-Duby R, Olson EN (2003) Control of cardiac growth and function by calcineurin signaling. J Biol Chem 278:36981–36984PubMedCrossRefGoogle Scholar
  12. 12.
    Rusnak F, Mertz P (2000) Calcineurin: form and function. Physiol Rev 80:1483–1521PubMedGoogle Scholar
  13. 13.
    Maillet M, Davis J, Auger-Messier M, York A, Osinska H, Piquereau J, Lorenz JN, Robbins J, Ventura-Clapier R, Molkentin JD (2010) Heart-specific deletion of CnB1 reveals multiple mechanisms whereby calcineurin regulates cardiac growth and function. J Biol Chem 285:6716–6724PubMedCrossRefGoogle Scholar
  14. 14.
    Liu L, Su Z, Xin S, Cheng J, Li J, Xu L, Wei Q (2012) The calcineurin B subunit (CnB) is a new ligand of integrin αM that mediates CnB-induced Apo2L/TRAIL expression in macrophages. J Immunol 188:238–247PubMedCrossRefGoogle Scholar
  15. 15.
    Su Z, Xin S, Xu L, Cheng J, Guo J, Li L, Wei Q (2012) The calcineurin B subunit induces TNF-related apoptosis-inducing ligand (TRAIL) expression via CD11b-NF-κB pathway in RAW264.7 macrophages. Biochem Biophys Res Commun 417:777–783PubMedCrossRefGoogle Scholar
  16. 16.
    Li J, Guo J, Su Z, Hu M, Liu W, Wei Q (2011) Calcineurin subunit B activates dendritic cells and acts as a cancer vaccine adjuvant. Int Immunol 23:327–334PubMedCrossRefGoogle Scholar
  17. 17.
    Su Z, Xin S, Li J, Guo J, Long X, Cheng J, Wei Q (2011) A new function for the calcineurin b subunit: antiplatelet aggregation and anticoagulation. IUBMB Life 63:1037–1044PubMedCrossRefGoogle Scholar
  18. 18.
    Li N, Zhang Z, Zhang W, Wei Q (2011) Calcineurin B subunit interacts with proteasome subunit alpha type 7 and represses hypoxia-inducible factor-1a activity via the proteasome pathway. Biochem Biophys Res Commun 405:468–472PubMedCrossRefGoogle Scholar
  19. 19.
    Simkhovich BZ, Marjoram P, Poizat C, Kedes L, Kloner RA (2003) Age-related changes of cardiac gene expression following myocardial ischemia/reperfusion. Arch Biochem Biophys 420:268–278PubMedCrossRefGoogle Scholar
  20. 20.
    Wu W, Jia Z, Liu P, Xie Z, Wei Q (2005) A novel PCR strategy for high-efficiency, automated site-directed mutagenesis. Nucleic Acids Res 33:e110PubMedCrossRefGoogle Scholar
  21. 21.
    Jiang G, Wei Q (2003) Function and structure of N-terminal and C-terminal domains of calcineurin B subunit. Biol Chem 384:1299–1303PubMedCrossRefGoogle Scholar
  22. 22.
    Kang PM, Haunstetter A, Aoki H, Usheva A, Izumo S (2000) Morphological and molecular characterization of adult cardiomyocyte apoptosis during hypoxia and reoxygenation. Circ Res 87:118–125PubMedCrossRefGoogle Scholar
  23. 23.
    Han Z, Hendrickson EA, Bremner TA, Wyche JH (1997) A sequential two-step mechanism for the production of the mature p17:p12 form of caspase-3 in vitro. J Biol Chem 272:13432–13436PubMedCrossRefGoogle Scholar
  24. 24.
    Hockenbery D, Nunez G, Milliman C, Schreiber RD, Korsmeyer SJ (1990) Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death. Nature 348:334–336PubMedCrossRefGoogle Scholar
  25. 25.
    Youle RJ, Strasser A (2008) The BCL-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol 9:47–59PubMedCrossRefGoogle Scholar
  26. 26.
    Whelan RS, Kaplinskiy V, Kitsis RN (2010) Cell death in the pathogenesis of heart disease: mechanisms and significance. Annu Rev Physiol 72:19–44PubMedCrossRefGoogle Scholar
  27. 27.
    Bueno OF, Lips DJ, Kaiser RA, Wilkins BJ, Dai YS, Glascock BJ, Klevitsky R, Hewett TE, Kimball TR, Aronow BJ, Doevendans PA, Molkentin JD (2004) Calcineurin Abeta gene targeting predisposes the myocardium to acute ischemia-induced apoptosis and dysfunction. Circ Res 94:91–99PubMedCrossRefGoogle Scholar
  28. 28.
    Pu WT, Ma Q, Izumo S (2003) NFAT transcription factors are critical survival factors that inhibit cardiomyocyte apoptosis during phenylephrine stimulation in vitro. Circ Res 92:725–731PubMedCrossRefGoogle Scholar
  29. 29.
    Kakita T, Hasegawa K, Iwai-Kanai E, Adachi S, Morimoto T, Wada H, Kawamura T, Yanazume T, Sasayama S (2001) Calcineurin pathway is required for endothelin-1-mediated protection against oxidant stress-induced apoptosis in cardiac myocytes. Circ Res 88:1239–1246PubMedCrossRefGoogle Scholar
  30. 30.
    De Windt LJ, Lim HW, Taigen T, Wencker D, Condorelli G, Dorn GW 2nd, Kitsis RN, Molkentin JD (2000) Calcineurin-mediated hypertrophy protects cardiomyocytes from apoptosis in vitro and in vivo: an apoptosis-independent model of dilated heart failure. Circ Res 86:255–263PubMedCrossRefGoogle Scholar
  31. 31.
    Saito S, Hiroi Y, Zou Y, Aikawa R, Toko H, Shibasaki F, Yazaki Y, Nagai R, Komuro I (2000) Beta-adrenergic pathway induces apoptosis through calcineurin activation in cardiac myocytes. J Biol Chem 275:34528–34533PubMedCrossRefGoogle Scholar
  32. 32.
    Chu CH, Tzang BS, Chen LM, Liu CJ, Tsai FJ, Tsai CH, Lin JA, Kuo WW, Bau DT, Yao CH, Huang CY (2009) Activation of insulin-like growth factor II receptor induces mitochondrial-dependent apoptosis through G(alpha)q and downstream calcineurin signaling in myocardial cells. Endocrinology 150:2723–2731PubMedCrossRefGoogle Scholar
  33. 33.
    Lotem J, Kama R, Sachs L (1999) Suppression or induction of apoptosis by opposing pathways downstream from calcium-activated calcineurin. Proc Natl Acad Sci USA 96:12016–12020PubMedCrossRefGoogle Scholar
  34. 34.
    Molkentin JD (2001) Calcineurin, mitochondrial membrane potential, and cardiomyocyte apoptosis. Circ Res 88:1220–1222PubMedCrossRefGoogle Scholar
  35. 35.
    Antonsson B (2004) Mitochondria and the Bcl-2 family proteins in apoptosis signaling pathways. Mol Cell Biochem 256(257):141–155PubMedCrossRefGoogle Scholar
  36. 36.
    Maulik N, Engelman RM, Rousou JA, Flack JE 3rd, Deaton D, Das DK (1999) Ischemic preconditioning reduces apoptosis by upregulating anti-death gene Bcl-2. Circulation 100:II369–II375PubMedCrossRefGoogle Scholar
  37. 37.
    Shibasaki F, Kondo E, Akagi T, McKeon F (1997) Suppression of signalling through transcription factor NF-AT by interactions between calcineurin and Bcl-2. Nature 386:728–731PubMedCrossRefGoogle Scholar
  38. 38.
    Wei Q, Lian ML, Jing FZ, Zhang N, Yan MS, Chen Y, Gao QS (2002) Studies of calcineurin B subunit from genetic engineering for use in medicine. Drug Dev Res 56:40–43CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2012

Authors and Affiliations

  • Junxia Guo
    • 1
    • 2
  • Shengquan Mi
    • 2
  • Jing Li
    • 1
  • Wei Liu
    • 1
  • Yanxia Yin
    • 1
  • Qun Wei
    • 1
  1. 1.Department of Biochemistry and Molecular Biology, Beijing Key Laboratory of Genetic Engineering Medicine and BiotechnologyBeijing Normal UniversityBeijingChina
  2. 2.Beijing Key Laboratory of Bioactive Substances and Functional FoodsCollege of Arts and Science of Beijing Union UniversityBeijingChina

Personalised recommendations