Molecular and Cellular Biochemistry

, Volume 369, Issue 1–2, pp 227–233 | Cite as

Pharmacological inhibition of GSK-3β produces late phase of cardioprotection in hyperlipidemic rat: possible involvement of HSP 72

  • Harlokesh Narayan YadavEmail author
  • Manjeet Singh
  • Pyare Lal Sharma


The acute, as well as late, phase of cardioprotection induced by ischemic preconditioning is abolished in hyperlipidemic (HL) rat heart. The pharmacological inhibition of glycogen synthase kinase-3β (GSK-3β), has earlier been reported to restore this attenuated acute cardioprotective effect. However, it not known whether GSK-3β inhibitors administered 24 h before the ischemic injury would restore the late cardioprotective in HL rat and, if yes, the role of heat shock protein 72 (HSP 72) in its modulation. Hyperlipidemia was produced in rat by feeding high-fat diet for 6 weeks. Isolated perfused rat heart was subjected to 30 min of ischemia followed by 120 min of reperfusion (I/R). Myocardial infarct size was estimated by triphenyltetrazolium chloride staining, while lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB) levels were analyzed from coronary effluent. GSK-3β inhibitors, SB 216763 (SB, 0.6 mg/kg, i.p.), and indirubin-3 monoxime (IND, 0.4 mg/kg, i.p.), administered 24 h before the isolation of heart, significantly decreased the I/R-induced myocardial infarct size and the release of LDH and CK-MB. The cardioprotective effect of GSK-3β inhibitors was significantly attenuated by quercetin (4 mg/kg, i.p.), a HSP 72 inhibitor, administered 1 h before the administration of SB or IND. That the late phase of cardioprotection induced by pretreatment with GSK-3β inhibitors is not attenuated/lost in HL rat heart is a new finding in our study. Our results indicate that HSP 72 acts on pathway of GSK-3β and plays a significant role in cardioprotection.


Glycogen synthase kinase-3β Ischemic preconditioning Hyperlipidemia 



No external source of funding was used in this study.


  1. 1.
    Murry CE, Jennings RB, Reimer KA (1986) Preconditioning with ischemia: a delay of lethal cell injury in ischemic myocardium. Circulation 74:1124–1136PubMedCrossRefGoogle Scholar
  2. 2.
    Juhaszaova M, Zorov DB, Kim SH, Pepe SFuQ, Fishbein KW, Ziman BD, Wang S, Ytrehus K, Antos CL, lson EN, Sollott SJ (2004) Glycogen synthase kinase-3beta mediates convergence of protection signaling to inhibit the mitochondrial permeability transition pore. J Clin Invest 113:1535–1549Google Scholar
  3. 3.
    Gross ER, Gross GJ (2006) Ligand triggers of classical preconditioning and postconditioning. Cardiovasc Res 70:212–221PubMedCrossRefGoogle Scholar
  4. 4.
    Downey JM, Davis AM, Cohen MV (2007) Signalling pathways in ischemic preconditioning. Heart Fail Rev 12:181–188PubMedCrossRefGoogle Scholar
  5. 5.
    Gross ER, Hsu AK, Gross GJ (2006) The JAK/STAT pathway is essential for opoid-induced cardioprotection: JAK2 as a mediator of STAT3, AKT, and GSK-3beta. Am J Physiol Heart Circ Physiol 291:H827–H834PubMedCrossRefGoogle Scholar
  6. 6.
    Baxter GF, Yellon DM (1999) ATP-sensitive K+ channels mediate the delayed cardioprotective effect of adenosine A1 receptor activation. J Mol Cell Cardiol 31:981–989PubMedCrossRefGoogle Scholar
  7. 7.
    Baxter GF, Ferdinandy P (2001) Delayed preconditioning of myocardium: current perspectives. Basic Res Cardiol 96:329–344PubMedCrossRefGoogle Scholar
  8. 8.
    Hutter MM, Sievers RE, Barbosa V, Wolfe CL (1994) 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 89:355–360PubMedCrossRefGoogle Scholar
  9. 9.
    Snoeckx IH, Cornelussen RN, Van Nieuwenhoven FA, Reneman RS, van der Vusse GJ (2001) Heat shock proteins and cardiovascular pathophysiology. Physiol Rev 81:1461–1497Google Scholar
  10. 10.
    William RS, Benjamin IJ (2000) Protective responses in the ischemic myocardium. J Clin Invest 106:813–818CrossRefGoogle Scholar
  11. 11.
    Zapletal C, Fallsehr C, Reidel M, Loffler T, Gebhard MM, Golling M (2010) Induction of HSP70 shows differences in protection against I/R injury derived by ischemic preconditioning and intermittent clamping. Microvasc Res 80(3):365–371PubMedCrossRefGoogle Scholar
  12. 12.
    Karmazyn M, Mailer K, Currie RW (1990) Acquisition and decay of heat-shock-enhanced post-ischemic ventricular recovery. Am J Physiol Heart Circ Physiol 259:H424–H431Google Scholar
  13. 13.
    Tranter M, Ren X, Forde T, Wilhide ME, Chen J, Sartor MA (2011) NF-kappaB driven cardioprotective gene programs; Hsp70.3 and cardioprotection after late ischemic preconditioning. J Mol Cell Cardiol 49(4):664–672CrossRefGoogle Scholar
  14. 14.
    Yamashita N, Hoshida S, Nishida M, Igarashi J, Aoki K, Hori M (1997) Time-course of tolerance to ischemia-reperfusion injury and induction of heat shock protein 72 by heat stress in the rat heart. J Mol Cell Cardiol 29:1815–1821PubMedCrossRefGoogle Scholar
  15. 15.
    Marber MS, Walker JM, Latchman DS, Yellon DM (1994) Myocardial protection after whole body heat stress in the rabbit is dependent on metabolic substrate and is related to the amount of the inducible 70-kD heat stress protein. J Clin Invest 93:1087–1094PubMedCrossRefGoogle Scholar
  16. 16.
    Yin C, Salloum FN, Kukreja RC (2009) A novel role of micro RNA in late preconditioning: upregulation of endothelial nitric oxide synthase and heat shock protein 70. Circ Res 104(5):572–575PubMedCrossRefGoogle Scholar
  17. 17.
    Gross ER, Hsu AK, Gross GJ (2008) Delayed cardioprotection afforded by the glycogen synthase kinase 3 inhibitor SB-216763 occurs via a KATP- and MPTP-dependent mechanism at reperfusion. Am J Physiol Heart Circ Physiol 294:H1497–H1500PubMedCrossRefGoogle Scholar
  18. 18.
    Csont T, Balogh G, Csonka C, Boros Imre, Horvath I (2002) Hyperlipidemia induced by high cholesterol diet inhibits heat shock response in rat hearts. Biochem Biophys Res Commun 290:1535–1538PubMedCrossRefGoogle Scholar
  19. 19.
    Ferdinandy P, Szilvassy Z, Baxter GF (1998) Adaptation to myocardial stress in disease states: is preconditioning a healthy heart phenomenon? Trends Pharmacol Sci 19:223–229PubMedCrossRefGoogle Scholar
  20. 20.
    Kyriakides ZS, Psychari S, Iliodromitis EK, Kolettis TM, Sbarouni E, Kremastinos DT (2002) Hyperlipidaemia prevents the expected reduction of myocardial ischemia on repeated balloon inflations during angioplasty. Chest 121:1211–1215PubMedCrossRefGoogle Scholar
  21. 21.
    Yadav HN, Singh M, Sharma PL (2010) Modulation of the cardioprotective effect of ischemic preconditioning in hyperlipidaemic rat heart. Eur J Pharmacol 643(1):78–83PubMedCrossRefGoogle Scholar
  22. 22.
    Chu B, Zhong R, Soncin F, Stevenson MA, Calderwood SK (1998) Transcriptional activity of heat shock factor 1 at 37 °C is repressed through phosphorylation on two distinct serine residues by glycogen synthase kinase 3 and protein kinases Cα and Cζ. J Biol Chem 273:18640–18646PubMedCrossRefGoogle Scholar
  23. 23.
    Pearce NJ, Arch JR, Clapham JC, Coghlan MP, Corcoran SL, Lister CA (2004) Development of glucose intolerance in male transgenic mice overexpressing human glycogen synthase kinase-3beta on a muscle-specific promoter. Metabolism 53:1322–1330PubMedCrossRefGoogle Scholar
  24. 24.
    Bijur GF, Jope RS (2000) Opposing actions of phospahtidylinositol 3-kinase and glycogen synthase kinase-3 beta in the regulation of HSF-1. Neurochemistry 75:2401–2408Google Scholar
  25. 25.
    Wang TD, Chen WJ, Mau TJ, Lin JW, Lin WW, Lee YT (2003) Attenuation of increased myocardial ischemia–reperfusion injury conferred by hypercholesterolemia through pharmacological inhibition in the caspase-1 cascade. Br J Pharmacol 138:291–300PubMedCrossRefGoogle Scholar
  26. 26.
    Lorkowska B, Bartus M, Franczyk M, Kostogrys RB, Jawien J, Pisulewski PM (2006) Hypercholesterolemia does not alter endothelial function in spontaneously hypertensive rats. J Pharmacol Exp Ther 317:1019–1026PubMedCrossRefGoogle Scholar
  27. 27.
    Reeves PG (1997) Components of the AIN-93 diets as improvements in the AIN-76a diet. J Nutr 127:838S–841SPubMedGoogle Scholar
  28. 28.
    Langendorff O (1895) Untersuchungen am uberlebender saugethierherzen. Pflügers Arch Gesmate Physiol 61:291–332CrossRefGoogle Scholar
  29. 29.
    Fishbein MC, Meerbaum S, Rit J, Lando U, Kanmatsuse K, Merair JC, Corday E, Ganz W (1981) Early phase acute myocardial infarct size quantification: validation of the triphenyltetrazolium chloride tissue enzyme staining technique. Am Heart J 101:593–600PubMedCrossRefGoogle Scholar
  30. 30.
    Chopra K, Singh M, Kaul N, Andrabi KJ, Ganguly NK (1992) Decrease of myocardial infarct size with dessferrioxamine. Possible role of oxygen free radicals in its ameliorative effect. Mol Cell Biochem 113:71–76PubMedCrossRefGoogle Scholar
  31. 31.
    Hosokawa N, Hirayoshi K, Nakai A, Hosokawa Y, Marui N, Yoshida M, Sakaki T, Nishino H, Aoike A, Kawai K, Nagata K (1990) Flavonoids inhibit the expression of heat shock proteins. Cell Struct Funct 15:393–401PubMedCrossRefGoogle Scholar
  32. 32.
    Nagai N, Nakai A, Nagata K (1995) Quercetin suppresses heat shock response by down regulation of HSF1. Biochem Biophys Res Commun 208:1099–1105PubMedCrossRefGoogle Scholar
  33. 33.
    Debes A, Oerding M, Willers R, Gobel U, Wessalowski R (2003) Sensitization of human Ewing’s tumor cells to chemotherapy and heat treatment by the bioflavonoid quercetin. Anticancer Res 23:3359–3366PubMedGoogle Scholar
  34. 34.
    Yadav HN, Singh M, Sharma PL (2010) Involvement of GSK-3β in attenuation of cardioprotective effect of ischemic preconditioning in diabetic rat heart. Mol Cell Biochem 343:75–81PubMedCrossRefGoogle Scholar
  35. 35.
    Bolli R (2000) The late phase of preconditioning. Cir Res 87:972–983CrossRefGoogle Scholar
  36. 36.
    Bolli R, Dawn B, Tang XL, Qiu Y, Ping P, Xuan YT (1998) The nitric oxide hypothesis of late preconditioning. Basic Res Cardiol 93:325–338PubMedCrossRefGoogle Scholar
  37. 37.
    Kis A, Yellon DM, Baxter GF (2003) Second window of protection following myocardial preconditioning: an essential role for PI3 kinase and p70s6 kinase. J Mol Cell Cardiol 35:1063–1071PubMedCrossRefGoogle Scholar
  38. 38.
    Latchman DS (2001) Heat shock proteins and cardiac protection. Cardiovasc Res 51:637–646PubMedCrossRefGoogle Scholar
  39. 39.
    Li R, Zheng W, Pi R, Gao J, Zhang H, Le PWK (2007) Activation of peroxisome proliferator-activated receptor-ά prevents glycogen synthase 3β phosphorylation and inhibits cardiac hypertrophy. FEBS 581:3311–3316CrossRefGoogle Scholar
  40. 40.
    Marban E (1991) Myocardial stunning and hibernation: the physiology behind the colloquialisms. Circulation 83:681–688PubMedCrossRefGoogle Scholar
  41. 41.
    Matter WF, Brown RF, Vlahos CJ (1992) The inhibition of phosphatidylinositol 3-kinase by quercetin and analogs. Biochem Biophys Res Commun 186:624–631PubMedCrossRefGoogle Scholar
  42. 42.
    Levy J, Teuerstein I, Marbach M, Radian S, Sharoni Y (1984) Tyrosine protein kinase activity in the DMBA-induced rat mammary tumor: inhibition by quercetin. Biochem Biophys Res Commun 123:1227–1233PubMedCrossRefGoogle Scholar
  43. 43.
    Yao YW, Zhang G, Zhang Y, Li W, Wang C, Yin CY, Zhang F (2011) Lipopolysaccharide pretreatment protects against ischemia/reperfusion injury via increase of HSP70 and inhibition of NF-κB. Cell Stress Chaperones 16:287–296PubMedCrossRefGoogle Scholar
  44. 44.
    Liu J, Kam KWL, Zhou J, Yan W, Chen M, Wu S, Wong TM (2004) Effects of heat shock protein 70 activation by metabolic inhibition preconditioning or к-opioid receptor stimulation on Ca2+ homeostasis in rat ventricular myocytes subjected to ischemic insults. J Pharmacol Exp Ther 310:606–613PubMedCrossRefGoogle Scholar
  45. 45.
    Malyshev IYu, Malysheva EV (1998) Heat-shock proteins and cardioprotection. Bull Exp Biol Med 126:1177–1183CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2012

Authors and Affiliations

  • Harlokesh Narayan Yadav
    • 1
    • 2
    Email author
  • Manjeet Singh
    • 1
  • Pyare Lal Sharma
    • 1
  1. 1.Department of PharmacologyI.S.F. College of PharmacyMogaIndia
  2. 2.Division of PharmacologyInstitute of Pharmaceutical Research, GLA UniversityMathuraIndia

Personalised recommendations