Advertisement

Novel functional role of heat shock protein 90 in protein kinase C-mediated ischemic postconditioning

Published:January 31, 2014DOI:https://doi.org/10.1016/j.jss.2014.01.038

      Abstract

      Background

      Previous studies have shown that heat shock protein 90 (HSP90) plays a vital role in ischemic preconditioning. The present study was designed to explore whether HSP90 might be responsible for cardioprotection in ischemic postconditioning (PostC).

      Materials and methods

      Rat hearts underwent 30 min of regional ischemia and 2 h of reperfusion in situ, and PostC was effected with three cycles of 30-s reperfusion and 30-s coronary artery occlusion at the end of ischemia. Ninety rats were randomized into five groups: sham; ischemia–reperfusion (I/R); PostC; 1 mg/kg HSP90 inhibitor geldanamycin (GA) plus PostC (PostC + GA1); and 5 mg/kg GA plus PostC (PostC + GA5). The GA was administered 10 min before reperfusion.

      Results

      Compared with the I/R group, the PostC group exhibited lower infarct size (46.7 ± 3.0% versus 27.4 ± 4.0%, respectively), release of lactate dehydrogenase and creatine kinase-MB (2252.6 ± 350.8 versus 1713.7 ± 202.4 IU/L, 2804.3 ± 315.7 versus 1846.2 ± 238.0 IU/L, respectively), cardiomyocyte apoptosis (48.4 ± 5.6% versus 27.6 ± 3.8%, respectively), and mitochondrial damage. These beneficial effects were accompanied by an increase in mitochondrial Bcl-2 levels and a decrease in Bax levels. In addition, mitochondrial protein kinase Cepsilon (PKCepsilon) was relatively low in the I/R group but significantly higher in the PostC group, whereas cytosolic PKCepsilon was relatively high in the I/R group but significantly lower in the PostC group, suggesting the translocation of PKCepsilon from cytosol to mitochondria during PostC. However, blocking HSP90 function with GA inhibited the protection of PostC and PKCepsilon mitochondrial translocation.

      Conclusions

      HSP90 is critical in PostC-induced cardioprotection, and its activity might be linked to mitochondrial targeting of PKCepsilon, the activation of which results in upregulation of its target gene, Bcl-2, and the inhibition of proapoptotic Bax in mitochondria.

      Keywords

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Journal of Surgical Research
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Zhao Z.Q.
        • Corvera J.S.
        • Halkos M.E.
        • et al.
        Inhibition of myocardial injury by ischemic postconditioning during reperfusion: comparison with ischemic preconditioning.
        Am J Physiol Heart Circ Physiol. 2003; 285: H579
        • Staat P.
        • Rioufol G.
        • Piot C.
        • et al.
        Postconditioning the human heart.
        Circulation. 2005; 112: 2143
        • Lonborg J.
        • Kelbaek H.
        • Vejlstrup N.
        • et al.
        Cardioprotective effects of ischemic postconditioning in patients treated with primary percutaneous coronary intervention, evaluated by magnetic resonance.
        Circ Cardiovasc Interv. 2010; 3: 34
        • Philipp S.
        • Yang X.M.
        • Cui L.
        • Davis A.M.
        • Downey J.M.
        • Cohen M.V.
        Postconditioning protects rabbit hearts through a protein kinase C adenosine A2b receptor cascade.
        Cardiovasc Res. 2006; 70: 308
        • Zatta A.J.
        • Kin H.
        • Lee G.
        • et al.
        Infarct-sparing effect of myocardial postconditioning is dependent on protein kinase C signalling.
        Cardiovasc Res. 2006; 70: 315
        • Dong S.
        • Teng Z.
        • Lu F.H.
        • et al.
        Post-conditioning protects cardiomyocytes from apoptosis via PKC(epsilon)-interacting with calcium-sensing receptors to inhibit endo(sarco)plasmic reticulum-mitochondria crosstalk.
        Mol Cell Biochem. 2010; 341: 195
        • Penna C.
        • Perrelli M.G.
        • Tullio F.
        • Angotti C.
        • Pagliaro P.
        Acidic infusion in early reperfusion affects the activity of antioxidant enzymes in postischemic isolated rat heart.
        J Surg Res. 2013; 183: 111
        • Kabir A.M.
        • Clark J.E.
        • Tanno M.
        • et al.
        Cardioprotection initiated by reactive oxygen species is dependent on activation of PKCε.
        Am J Physiol Heart Circ Physiol. 2006; 291: 1893
        • Baines C.P.
        • Zhang J.
        • Wang G.-W.
        • et al.
        Mitochondrial PKCepsilon and MAPK form signaling modules in the murine heart: enhanced mitochondrial PKCepsilon-MAPK interactions and differential MAPK activation in PKCepsilon-induced cardioprotection.
        Circ Res. 2002; 90: 390
        • Liu G.S.
        • Cohen M.V.
        • Mochly-Rosen D.
        • Downey J.M.
        Protein kinase C-epsilon is responsible for the protection of preconditioning in rabbit cardiomyocytes.
        J Mol Cell Cardiol. 1999; 31: 1937
        • Jabůrek M.
        • Costa A.D.
        • Burton J.R.
        • Costa C.L.
        • Garlid K.D.
        Mitochondrial PKC epsilon and mitochondrial ATP-sensitive K+ channel copurify and coreconstitute to form a functioning signaling module in proteoliposomes.
        Circ Res. 2006; 99: 878
        • Sun H.Y.
        • Wang N.P.
        • Halkos M.
        • et al.
        Postconditioning attenuates cardiomyocyte apoptosis via inhibition of JNK and p38 mitogen-activated protein kinase signaling pathways.
        Apoptosis. 2006; 11: 1583
        • Tian Y.
        • Zhang W.
        • Xia D.
        • Modi P.
        • Liang D.
        • Wei M.
        Postconditioning inhibits myocardial apoptosis during prolonged reperfusion via a JAK2-STAT3-Bcl-2 pathway.
        J Biomed Sci. 2011; 18: 53
        • Yu B.
        • Song B.
        Notch 1 signalling inhibits cardiomyocyte apoptosis in ischaemic postconditioning.
        Heart Lung Circ. 2014; 23: 152
        • Husainy M.A.
        • Dickenson J.M.
        • Galiñanes M.
        The MPTP status during early reoxygenation is critical for cardioprotection.
        J Surg Res. 2012; 174: 62
        • Jiao J.D.
        • Garg V.
        • Yang B.
        • Hu K.
        Novel functional role of heat shock protein 90 in ATP-sensitive K+ channel-mediated hypoxic preconditioning.
        Cardiovasc Res. 2008; 77: 126
        • Harrison E.M.
        • Sharpe E.
        • Bellamy C.O.
        • et al.
        Heat shock protein 90-binding agents protect renal cells from oxidative stress and reduce kidney ischemia-reperfusion injury.
        Am J Physiol Renal Physiol. 2008; 295: F397
        • Yang Z.
        • Sun W.
        • Hu K.
        Molecular mechanism underlying adenosine receptor-mediated mitochondrial targeting of protein kinase C.
        Biochim Biophys Acta. 2012; 1823: 950
        • Bradford M.M.
        A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
        Anal Biochem. 1976; 7: 248
        • Argaud L.
        • Gateau-Roesch O.
        • Augeul L.
        • et al.
        Increased mitochondrial calcium coexists with decreased reperfusion injury in postconditioned (but not preconditioned) hearts.
        Am J Physiol Heart Circ Physiol. 2008; 294: H386
        • Okorie M.I.
        • Bhavsar D.D.
        • Ridout D.
        • et al.
        Postconditioning protects against human endothelial ischaemia–reperfusion injury via subtype-specific KATP channel activation and is mimicked by inhibition of the mitochondrial permeability transition pore.
        Eur Heart J. 2011; 32: 1266
        • Zhou C.
        • Yao Y.
        • Zheng Z.
        • et al.
        Stenting technique, gender, and age are associated with cardioprotection by ischaemic postconditioning in primary coronary intervention: a systematic review of 10 randomized trials.
        Eur Heart J. 2012; 33: 3070
        • Young J.C.
        • Hoogenraad N.J.
        • Hartl F.U.
        Molecular chaperones Hsp90 and Hsp70 deliver preproteins to the mitochondrial import receptor Tom70.
        Cell. 2003; 112: 41
        • Rodriguez-Sinovas A.
        • Boengler K.
        • Cabestrero A.
        • et al.
        Translocation of connexin 43 to the innermitochondrial membrane of cardiomyocytes through the heat shock protein 90-dependent TOM pathway and its importance for cardioprotection.
        Circ Res. 2006; 99: 93
        • Budas G.R.
        • Churchill E.N.
        • Disatnik M.H.
        • Sun L.
        • Mochly-Rosen D.
        Mitochondrial import of PKCepsilon is mediated by HSP90: a role in cardioprotection from ischaemia and reperfusion injury.
        Cardiovasc Res. 2010; 88: 83
        • Körner C.
        • Keklikoglou I.
        • Bender C.
        • Wörner A.
        • Münstermann E.
        • Wiemann S.
        MicroRNA-31 sensitizes human breast cells to apoptosis by direct targeting of protein kinase C epsilon (PKCepsilon).
        Biol Chem. 2013; 288: 8750
        • Shankar E.
        • Krishnamurthy S.
        • Paranandi R.
        • Basu A.
        PKCepsilon induces Bcl-2 by activating CREB.
        Int J Oncol. 2010; 36: 883
        • Gray M.O.
        • Zhou H.Z.
        • Schafhalter-Zoppoth I.
        • Zhu P.
        • Mochly-Rosen D.
        • Messing R.O.
        Preservation of base-line hemodynamic function and loss of inducible cardioprotection in adult mice lacking protein kinase C epsilon.
        J Biol Chem. 2004; 279: 3596
        • Inagaki K.
        • Churchill E.
        • Mochly-Rosen D.
        Epsilon protein kinase C as a potential therapeutic target for the ischemic heart.
        Cardiovasc Res. 2006; 70: 222
        • Steinberg R.
        • Harari O.A.
        • Lidington E.A.
        • et al.
        A protein kinase Cε-anti-apoptotic kinase signaling complex protects human vascular endothelial cells against apoptosis through induction of Bcl-2.
        J Biol Chem. 2007; 282: 32288
        • Zhu L.
        • Yu Y.
        • Chua B.H.
        • Ho Y.S.
        • Kuo T.H.
        Regulation of sodium-calcium exchange and mitochondrial energetics by Bcl-2 in the heart of transgenic mice.
        J Mol Cell Cardiol. 2001; 33: 2135
        • Murphy E.
        • Steenbergen C.
        Preconditioning: the mitochondrial connection.
        Annu Rev Physiol. 2007; 69: 51