Apoptosis

, Volume 13, Issue 2, pp 305–317 | Cite as

Insulin inhibits β-adrenergic action in ischemic/reperfused heart: a novel mechanism of insulin in cardioprotection

Original Paper

Abstract

Objective

Sympathetic overactivity is closely connected with cell injury and contractile dysfunction during myocardial ischemia/reperfusion (MI/R). Insulin exerts protection for the I/R heart and the underlying mechanisms remain unclear. This study aimed to investigate the ability of insulin to modulate β-adrenergic actions on myocardial contraction and post-ischemic injury in acute MI/R and the underlying mechanism.

Methods

Isolated hearts from adult SD rats were subjected to MI/R (30 min/2 h) and treated with isoproterenol (ISO) or/and insulin. Myocardial contraction, cardiomyocyte apoptosis, myocardial injury and infarction were assessed. In a separate study, isolated ventricular myocytes were subjected to simulated I/R (15/30 min) and myocyte shortening and intracellular Ca2+ transient in response to ISO during reperfusion were assessed with presence or absence of insulin.

Results

In isolated I/R hearts, insulin largely reversed the ISO-associated contractile functional impairment at 2 h after MI/R, inhibiting ISO-induced declines in heart rate and left ventricular systolic pressure by 34.0% and 23.0% and preventing ISO-induced elevation in left ventricular end-diastolic pressure by 28.7% respectively (all < 0.05). In addition, ISO alone resulted in enlarged infarct size, elevated CK and LDH activity and increased apoptotic index in I/R hearts compared with vehicle, which were inhibited by treatment of insulin (all < 0.05). Interestingly, in SI/R cardiomyocytes, insulin alone at 10−7 mol/l increased cell contraction whereas attenuated the positive inotropic response to ISO (10−9 mol/l) during R as evidenced by a 18.7% reduction in peak twitch amplitude and a 23.9% reduction in calcium transient amplitude (both < 0.05). Moreover, insulin blunted ISO-mediated increase in PKA activity, enhanced the PKA-dependent phosphorylation of phospholamban (PLB), resulting in increased sarcoplasmic reticulum Ca2+-ATPase (SERCA2a) activity.

Conclusion

Insulin attenuated the contractile response to β-AR stimulation and suppressed ISO-elicited cardiac dysfunction and cell injury in MI/R. The inhibitory effect of insulin on the β-adrenergic action involved the inhibition of PKA-mediated Ca2+ transient and promotion of post-ischemic Ca2+ handling.

Keywords

β-Adrenoceptor Insulin Ischemia and reperfusion injury Apoptosis PKA 

References

  1. 1.
    Mak KH, Topol EJ (2000) Emerging concepts in the management of acute myocardial infarction in patients with diabetes mellitus. J Am Col Cardiol 35:563–568CrossRefGoogle Scholar
  2. 2.
    Vadlamudi RV, McNeill JH (1984) Effect of experimental diabetes on isolated rat heart responsiveness to isoproterenol. Can J Physiol Pharmacol 62:124–131PubMedGoogle Scholar
  3. 3.
    Heyliger CE, Pierce GN, Singal PK, Beamish RE, Dhalla NS (1982) Cardiac α- and β-adrenergic receptor alterations in diabetic cardiomyopathy. Basic Res Cardiol 77:610–618PubMedCrossRefGoogle Scholar
  4. 4.
    Nishio Y, Kashiwagi A, Kida Y et al (1988) Deficiency of cardiac β-adrenergic receptor in streptozotocin-induced diabetic rats. Diabetes 37:1181–1187PubMedCrossRefGoogle Scholar
  5. 5.
    Wichelhaus A, Russ M, Petersen S, Eckel J (1994) G-protein expression and adenylate cyclase regulation in ventricular cardiomyocytes from STZ-diabetic rats. Am J Physiol 267:H548–H555PubMedGoogle Scholar
  6. 6.
    Fein FS (1990) Diabetic cardiomyopathy. Diabetes Care 13:1169–1179PubMedCrossRefGoogle Scholar
  7. 7.
    Pfaffman MA (1980) The effects of streptozotocin-induced diabetes and insulin-treatment on the cardiovascular system of the rat. Res Commun Chem Pathol Pharmacol 28:27–41PubMedGoogle Scholar
  8. 8.
    Schömig A, Dart AM, Dietz R, Mayer E, Kubler W (1984) Release of endogenous catecholamines in the ischemic myocardium of the rat. Part A: locally mediated release. Circ Res 55:689–701PubMedGoogle Scholar
  9. 9.
    Lameris TW, de Zeeuw S, Alberts G et al (2000) Time course and mechanism of myocardial catecholamine release during transient ischemia in vivo. Circulation 101:2645–2650PubMedGoogle Scholar
  10. 10.
    Lubbe WF, Podzuweit T, Opie LH (1992) Potential arrhythmogenic role of cyclic adenosine monophosphate (AMP) and cytosolic calcium overload: implications for prophylactic effects of beta-blockers in myocardial infarction and proarrhythmic effects of phosphodiesterase inhibitors. J Am Coll Cardiol 19:1622–1633PubMedCrossRefGoogle Scholar
  11. 11.
    Gao F, Gao E, Yue TL et al (2002) Nitric oxide mediates the antiapoptotic effect of insulin in myocardial ischemia-reperfusion: the roles of PI3-kinase, Akt, and endothelial nitric oxide synthase phosphorylation. Circulation 105:1497–1502PubMedCrossRefGoogle Scholar
  12. 12.
    Ma H, Zhang HF, Yu L et al (2006) Vasculoprotective effect of insulin in the ischemic/reperfused canine heart: role of Akt-stimulated NO production. Cardiovasc Res 69:57–65PubMedCrossRefGoogle Scholar
  13. 13.
    Zhang HF, Fan Q, Qian XX et al (2004) Role of insulin in the anti-apoptotic effect of glucoseinsulin-potassium in rabbits with acute myocardial ischemia and reperfusion. Apoptosis 9:777–783PubMedCrossRefGoogle Scholar
  14. 14.
    Mattiazzi A, Mundiña-Weilenmann C, Guoxiang C, Vittone L, Kranias E (2005) Role of phospholamban phosphorylation on Thr17 in cardiac physiological and pathological conditions. Cardiovasc Res 68:366–375PubMedCrossRefGoogle Scholar
  15. 15.
    Temsah RM, Dyck C, Netticadan T, Chapman D, Elimban V, Dhalla NS (2000) Effect of beta-adrenoceptor blockers on sarcoplasmic reticular function and gene expression in the ischemic-reperfused heart. J Pharmacol Exp Ther 293:15–23PubMedGoogle Scholar
  16. 16.
    Sande JB, Sjaastad I, Hoen IB et al (2002) Reduced level of serine(16) phosphorylated phospholamban in the failing rat myocardium: a major contributor to reduced SERCA2 activity. Cardiovasc Res 53:382–391PubMedCrossRefGoogle Scholar
  17. 17.
    Zucchi R, Ronca-Testoni S, Di Napoli P et al (1996) Sarcoplasmic reticulum calcium uptake in human myocardium subjected to ischemia and reperfusion during cardiac surgery. J Mol Cell Cardiol 28:1693–1701PubMedCrossRefGoogle Scholar
  18. 18.
    Krause SM, Jacobus WE, Becker LC (1989) Alterations in cardiac sarcoplasmic reticulum calcium transport in the postischemic “stunned” myocardium. Circ Res 65:526–530PubMedGoogle Scholar
  19. 19.
    Temsah RM, Netticadan T, Chapman D, Takeda S, Mochizuki S, Dhalla NS (1999) Alterations in sarcoplasmic reticulum function and gene expression in ischemic-reperfused rat heart. Am J Physiol 277:H584–594PubMedGoogle Scholar
  20. 20.
    Zhang B, Zhang HF, Fan Q, Ma XL, Gao F (2003) Insulin improves cardiac myocytes contractile function recovery in simulated ischemia-reperfusion: key role of Akt. Chin Sci Bull 48:1364–1369CrossRefGoogle Scholar
  21. 21.
    Yu J, Zhang HF, Wu F et al (2006) Insulin improves cardiomyocyte contractile function through enhancement of SERCA2a activity in simulated ischemia/reperfusion. Acta Pharmacol Sin 27:919–926PubMedCrossRefGoogle Scholar
  22. 22.
    Ma XL, Kumar S, Gao F et al (1999) Inhibition of p38 mitogenactivated protein kinase decreases cardiomyocyte apoptosis and improves cardiac function after myocardial ischemia and reperfusion. Circulation 99:1685–1691PubMedGoogle Scholar
  23. 23.
    Zhang QJ, Li QX, Zhang HF et al (2007) Swim training sensitizes myocardial response to insulin: role of akt-dependent eNOS activation. Cardiovasc Res 75:369–380PubMedCrossRefGoogle Scholar
  24. 24.
    Hopkins TA, Dyck JR, Lopaschuk GD (2003) AMP-activated protein kinase regulation of fatty acid oxidation in the ischaemic heart. Biochem Soc Trans 31:207–212PubMedCrossRefGoogle Scholar
  25. 25.
    McCormack J, Barr R, Wolff A, Lopaschuk G (1996) Ranolazine stimulates glucose oxidation in normoxic, ischemic, and reperfused ischemic rat hearts. Circulation 93:135–142PubMedGoogle Scholar
  26. 26.
    Remondino A, Kwon SH, Communal C et al (2003) Beta-adrenergic receptor-stimulated apoptosis in cardiac myocytes is mediated by reactive oxygen species/c-Jun NH2-terminal kinase-dependent activation of the mitochondrial pathway. Circ Res 92:136–138PubMedCrossRefGoogle Scholar
  27. 27.
    Hu A, Jiao X, Gao E et al (2006) Chronic beta-adrenergic receptor stimulation induces cardiac apoptosis and aggravates myocardial ischemia/reperfusion injury by provoking inducible nitric-oxide synthase-mediated nitrative stress. J Pharmacol Exp Ther 318(2):469–475PubMedCrossRefGoogle Scholar
  28. 28.
    Engfeldt P, Hellmér J, Wahrenberg H, Arner P (1988) Effects of insulin on adrenoceptor binding and the rate of catecholamine-induced lipolysis in isolated human fat cells. J Biol Chem 263(30):15553–15560PubMedGoogle Scholar
  29. 29.
    Karoor V, Baltensperger K, Paul H, Czech MP, Malbon CC (1995) Phosphorylation of tyrosyl residues 350/354 of the beta-adrenergic receptor is obligatory for counterregulatory effects of insulin. J Biol Chem 270:25305–25308PubMedCrossRefGoogle Scholar
  30. 30.
    Lee JC, Downing SE (1976) Effects of insulin on cardiac muscle contraction and responsiveness to norepinephrine. Am J Physiol 230:1360–1365PubMedGoogle Scholar
  31. 31.
    Austin C, Chess-Williams R (1994) The in-vitro effects of insulin and the effects of acute fasting on cardiac beta-adrenoceptor responses in the short-term streptozotocin-diabetic rat. J Pharm Pharmacol 46:326–331PubMedGoogle Scholar
  32. 32.
    Ferrara N, Abete P, Corbi G, Paolisso G (2005) Insulin-induced changes in β-adrenergic response: an experimental study in the isolated rat papillary muscle. Am J Hypertension 18:348–353CrossRefGoogle Scholar
  33. 33.
    Ren J, Walsh MF, Hamaty M, Sowers JR, Brown RA (1999) Augmentation of the inotropic response to insulin in diabetic rat hearts. Life Sci 65:369–380PubMedCrossRefGoogle Scholar
  34. 34.
    Lohse MJ (1993) Molecular mechanisms of membrane receptor desensitization. Biochem Biophys Acta 1179:171–188PubMedCrossRefGoogle Scholar
  35. 35.
    Strasser RH, Marquetant R, Kübler W (1990) Independent sensitization of beta-adrenoceptors and adenylate cyclase in acute myocardial ischaemia. Br J Clin Pharmacol 30(Suppl 1):27S–35SPubMedGoogle Scholar
  36. 36.
    Sichelschmidt OJ, Hahnefeld C, Hohlfeld T, Herberg FW, Schrör K (2003) Trapidil protects ischemic hearts from reperfusion injury by stimulating PKAII activity. Cardiovasc Res 58:602–610PubMedCrossRefGoogle Scholar
  37. 37.
    Singh RB, Chohan PK, Dhalla NS, Netticadan T (2004) The sarcoplasmic reticulum proteins are targets for calpain action in the ischemic-reperfused heart. J Mol Cell Cardiol 37(1):101–110PubMedCrossRefGoogle Scholar
  38. 38.
    Gavi S, Yin D, Shumay E, Wang HY, Malbon CC (2007) Insulin-like growth factor-I provokes functional antagonism and internalization of beta1-adrenergic receptors. Endocrinology 148(6):2653–2662PubMedCrossRefGoogle Scholar
  39. 39.
    Braz JC, Gregory K, Pathak A et al (2004) PKC-α regulates cardiac contractility and propensity toward heart failure. Nat Med 10:248–254PubMedCrossRefGoogle Scholar
  40. 40.
    Vittone L, Mundiña-Weilenmann C, Said M, Mattiazzi A (1998) Mechanisms involved in the acidosis enhancement of the isoproterenol-induced phosphorylation of phospholamban in the intact heart. J Biol Chem 273:9804–9811PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  1. 1.Departments of Cardiology and Physiology, Xijing HospitalFourth Military Medical UniversityXi’anChina
  2. 2.Department of Cardiology, Xijing HospitalFourth Military Medical UniversityXi’anChina
  3. 3.Department of PhysiologyFourth Military Medical UniversityXi’anChina

Personalised recommendations