Abstract
Heat stress (HS) and the subsequent expression of 72 kDa heat shock protein (HSP 72) has been shown to enhance post-ischemic functional recovery and reduce infarct size. Because the synthesis of heat shock proteins involves activation of heat shock transcription factors through phosphorylation, we hypothesized that inhibition of protein kinase C (PKC) would block HS mediated protection and expression of HSP 72 in the heart. Five groups of rats were studied (1) Sham anesthetized, (2) HS group - animals were heat shocked by raising the whole body core temperature to 42°C for 15 min, (3) Vehicle group - HS rats treated with 50% DMSO in saline, (4) PKC inhibitor-treated group - specific PKC antagonist, chelerythrine chloride (5 mg/kg, i.p) given 30 min prior to HS and (5) Vehicle treated control - non-HS rats treated with vehicle prior to ischemia/reperfusion. Hearts were subjected to 30 min of regional ischemia and 90 min of reperfusion 24 h after HS. Risk area was delineated by injection of 10% Evan's blue and infarct size determined using computer morphometry of tetrazolium stained sections. Infarct size (% area at risk) reduced significantly from 49.4 ± 2.3% (n = 7) in sham to 10.0 ± 2.5% (p < 0.01) and 9.1 ± 3.0% in HS and vehicle treated HS groups respectively (p < 0.05) Treatment with chelerythrine prior to HS increased infarct size to 49.4 ± 2.3% (p < 0.05). Infarct size in chelerythrine-treated non-HS ischemic/reperfused heart was 40.7 ± 5.4%, which did not differ significantly from vehicle-treated sham group. Western blot analysis demonstrated marked increase in HSP 72 in HS groups (with or without vehicle treatment) and pretreatment with chelerythrine chloride failed to inhibit the expression of HSP 72. The results suggest that HS-induced ischemic tolerance is mediated via PKC pathway and this protection does not appear to be directly related to the expression of HSP 72 in rat heart.
Similar content being viewed by others
References
Currie RW, Ross BM, David TA: Induction of the heat shock response in rats modulate heart rat, creatine kinase and protein synthesis after subsequent hyperthermic treatment. Cardiovasc Res 14: 87–93, 1990
Currie RW, Tanguay RM, Kingma JG Jr: Heat-shock response and limitation of tissue necrosis during occlusion/reperfusion in rabbit hearts. Circulation 87: 963–971, 1993
Hutter MW, Sievers RE, Barbosa V, Wolfe CL: Heat-shock protein induction in rat hearts: A direct correlation between the amount of heat-shock protein induced and the degree of myocardial protection. Circulation 89: 355–360, 1994
Shipley JB, Qian Y, Levasseur JE, Kukreja RC: Expression of the stress proteins HSP-27 and HSP-72 in rat hearts does not correlate with ischemic tolerance after heat shock. Circulation 92: (abstr) 1–654, 1995
Nayeem MA, Hess ML, Qian Y, Loesser KE, Kukreja RC: Delayed preconditioning of cultured adult rat cardiac myocytes. Role of 70 and 90 kD heat stress proteins in protection against lethal cellular injury. Am J Physiol 42: H861–H868, 1997
Marber MS, Mestril R, Chi SH, Sayen MR, Yellon DM: A heat shock protein 70 transgene results in myocardial protection. J Clin Invest 95: 1446–1556, 1995
Plumier JL, Ross BM, Currie RW, Angelidis CE, Kaziaris H, Kollias G, Pagoulatos GN: Transgenic mice expressing the human heat shock protein 70 have improved post-ischemic myocardial recovery. J Clin Invest 95: 1854–1860, 1995
Qian Y, Shipley JS, Levasseur JE, Kukreja RC: Expression of 72 and 27 kDa heat shock proteins does not correlate with ischemic tolerance by whole body hyperthermia in rat heart. J Mol Cell Cardiol 30: 1163–1172, 1998
Xi L, Chelliah J, Nayeem MA, Levasseur JE, Kukreja RC: Whole body heat shock fails to protect mouse heart against ischemia/reperfusion injury in an isolated perfused heart model. J Mol Cell Cardiol 29: 1998 (in press)
Gallo GJ, Schuetz TJ, Kingston RE: Regulation of heat shock factor in Schizosaccharomyces pombe more closely resembles regulation in mammals than in Saccharomyces cerevisiae. Mol Cell Biol 11: 281–288, 1991
Hensold JO, Hunt CR, Calderwood SK, Housman DE, Kingston RE: DNA binding of heat shock factor to the heat shock element is insufficient for transcriptional activation in murine erythroleukemia cells. Mol Cell Biol 10: 1600–1608, 1990
Sorger PK, Pelham RB: Yeast heat shock factor is an essential DNA-binding protein that exhibits temperature-dependent phosphorylation. Cell 54: 855–864, 1988
Cotto JJ, Kline M, Morimoto RI: Activation of heat shock factor 1 DNA binding precedes stress-induced serine phosphorylation. Evidence for a multistep pathway of regulation. J Biol Chem 271: 3355–3358, 1996
Choi HS, Li B, Lin Z, Huang E, Liu AY: cAMP and cAMP-dependent protein kinase regulate the human heat shock protein 70 gene promoter activity. J Biol Chem 266: 11858–11865, 1991
Hug H, Sarre TF: Protein kinase C isozymes: Divergence in signal transduction. Biochem J 291: 329–343, 1993
Ytrehus K, Liu Y, Downey JM: Preconditioning protects the ischemic rabbit heart by protein kinase C activation. Am J Physiol 266: H1145–H1152, 1994
Liu Y, Ytrehus K, Downey JM: Evidence that translocation of protein kinase C is a key event during ischemic preconditioning of rabbit myocardium. J Mol Cell Cardiol 26: 661–668, 1994
Armstrong SC, Hoover DB, Delacey MH, Ganote CE: Translocation of PKC, protein phosphatase inhibition and preconditioning of rabbit cardiomyocytes. J Mol Cell Cardiol 28: 1479–1492, 1996
Baxter GF, Goma FM, Yellon DM: Involvement of PKC in the delayed cytoprotection following sublethal ischemia in rabbit myocardium. Br J Pharmacol 115: 222–224, 1995
Kukreja RC, Qian Y, Flaherty EE: Protein kinase C is involved in heat stress-induced myocardial protection of rat heart. Circulation 94(suppl): 1–423, 1996
Qian YZ, Levasseur JE, Yoshida KI, Kukreja RC: KATP channels in rat heart: Blockade of ischemic and acetylcholine-mediated preconditioning by glibenclamide. Am.J.Physiol. 271: H23–H28, 1996
Mitchell MB, Meng X, Ao L, Brown JM, Harken AH, Banerjee A: Preconditioning of isolated rat heart is mediated by protein kinase C. Circ Res 76: 73–81, 1995
Wilson S, Song W, Karoly K, Ravingerova T, Vegh A, Papp J, Tomisawa S, Parratt JR, Pyne NJ: Delayed cardioprotection is associated with the sub-cellular relocalisation of ventricular protein kinase C epsilon, but not p42/44MAPK. Mol Cell Biochem 160–161: 225–230, 1996
Herbert JM, Augereau JM, Gieye J, Maffrand JD: Chelerythrine is a potent and specific inhibitor of PKC. Biochem Biophys Res Commun 172: 993–999, 1990
Komuro I, Yamazaki T, Katoh Y, Tobe K, Kadowaki T, Nagai R, Yazaki Y: Protein kinase cascade activated by mechanical stress in cardiocytes: Possible involvement of angiotensin II. Eur Heart J 16(suppl C): 8–11, 1995
Yao A, Takahashi T, Aoyagi T, Kinugawa K, Kohmoto O, Sugiura S, Serizawa T: Immediate-early gene induction and MAP kinase activation during recovery from metabolic inhibition in cultured cardiac myocytes. J Clin Invest 96: 69–77, 1995
Ghosh S, Baltimore D: Activation in vitro of NF-kappa B by phosphorylation of its inhibitor 1 kappa B. Nature 344: 678–682, 1990
Maggirwar SB, Dhanraj DN, Somani SM, Ramkumar V: Adenosine acts as an endogenous activator of the cellular antioxidant defense system. Biochem Biophys Res Comm 201: 508–515, 1994
Kukreja RC, Hess ML: Free Radicals, Cardiovascular Dysfunction and Protection Strategies. Landes Company, Austin, TX, 1994
Hunt C, Morimoto RI: Conserved features of eukaryotic HSP 70 genes revealed by comparison with the nucleotide sequence of human HSP 70. Proc Natl Acad Sci USA 82: 6455–6459, 1985
Lindquist S, Craig EA: The heat shock proteins. Ann Rev Genet 22: 631–637, 1988
Morimoto RI, Mosser D, McClanahan TK, Theodorakis NG, Williams G: Stress-Induced Proteins. Alan R. Liss Inc., New York, 1993
Hightower LE: Heat shock, stress proteins, chaperones, and proteotoxicity. Cell 66: 191–197, 1991
Kontos MC, Shipley JS, Kukreja RC: Heat stress improves functional recovery and induces synthesis of 27-and 70-kDa heat shock proteins without preserving sarcoplasmic reticulum function in the ischemic rat heart. J Mol Cell Cardiol 28: 1885–1894, 1996
Poteat HT, Chen FY, Kadison P, Sodroski JG, Haseltine WA: Protein kinase A-dependent binding of a nuclear factor to the 21-base-pair repeat of the human T-cell leukemia virus type 1 long terminal repeat. J Virol 64: 1264–1270, 1990
Manak JR, de Bisschop N, Kris RM, Prywes R: Casein kinase II enhances the DNA binding activity of serum response factor. Genes Dev 4: 955–967, 1990
Jacquier-Sarlin MR, Jornot L, Polia BS: Differential expression and regulation of hsp70 and hsp90 by phorbol esters and heat shock. J Biol Chem 270: 14094–14099, 1995
Wall SR, Fliss H, Korecky B: Role of catalase in myocardial protection against ischemia in heat shocked rats. Mol Cell Biochem 129: 187–194, 1993
Chelliah J, Xi L, Okubo S, Kukreja RC: Expression of HSP72 and antioxidant enzymes in mouse heart following whole body hyperthermia. J Mol Cell Cardiol 29 (abstr) (suppl): 157, 1997
Joannidis M, Cantley LG, Spokes K, Medina R, Pullman J, Rosen S, Epstein FH: Induction of heat-shock proteins does not prevent renal tubular injury following ischemia. Kidney Int 47: 1752–1759, 1995
Narasimhan P, Swanson RA, Sagar SM, Sharp FR: Astrocyte survival and HSP70 heat shock protein induction following heat shock and acidosis. Glia 17: 147–159, 1996
DeWeille JR, Schmid-Antomarchi H, Fosset M, Lazdunski M: Regulation of ATP-sensitive K+ channels in insulinoma cells: Activation by somatostatin and protein kinase C and the role of cAMP. Proc Natl Acad Sci USA 86: 2971–2975, 1989
Hu KL, Duan DY, Li GR, Nattel S: Protein kinase C activates ATP-sensitive K+ current in human and rabbit ventricular myocytes. Circ Res 78: 492–498, 1996
Nichols CG, Ripoll C, Lederer WJ: ATP-sensitive potassium channel modulation of guinea pig ventricular action potential and contraction. Circ Res 68: 280–287, 1991
Hoag JB, Qian Y, Nayeem MA, D'Angelo M, Kukreja RC: ATPsensitive potassium channel mediates delayed ischemic protection by heat stress in rabbit heart. Am J Physiol 42: H861–H868, 1997
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kukreja, R.C., Qian, YZ., Okubo, S. et al. Role of protein kinase C and 72 kDa heat shock protein in ischemic tolerance following heat stress in the rat heart. Mol Cell Biochem 195, 123–131 (1999). https://doi.org/10.1023/A:1006977311448
Issue Date:
DOI: https://doi.org/10.1023/A:1006977311448