Abstract
Increased protein kinase C (PKC) activity has been implicated in the pathogenesis of a number of diabetic complications, and high concentrations of glucose have been shown to increase PKC activity. The present study was designed to examine the role of PKC in diabetes-induced (and glucose-induced) cardiomyocyte dysfunction and insulin resistance (measured by glucose uptake). Adult rat ventricular myocytes were isolated from nondiabetic and type 1 diabetic animals (4–5 days post-streptozotocin treatment), and maintained overnight, with/without the nonspecific PKC inhibitor chelerythrine (CHEL = 1 μM). Myocyte mechanical properties were evaluated using a video edge-detection system. Basal and insulin-stimulated glucose uptake was measured with [3H]-2-deoxyglucose. Blunted insulin-stimulated glucose uptake was apparent in diabetic myocytes, and both mechanical dysfunctions (e.g., slowed shortening/relengthening) and insulin resistance were maintained in culture, and normalized by CHEL. Cardiomyocytes isolated from nondiabetic animals were cultured in a high concentration of glucose (HG = 25.5 mM) medium, with/without CHEL. HG myocytes exhibited slowed shortening/relengthening and impaired insulin-stimulated glucose uptake compared to myocytes cultured in normal glucose (5.5 mM), and both impairments were prevented by culturing cells in CHEL. Our data support the view that PKC activation contributes to both diabetes-induced abnormal cardiomyocyte mechanics and insulin resistance, and that elevated glucose is sufficient to induce these effects. (Mol Cell Biochem 262: 155–163, 2004)
Similar content being viewed by others
References
Grundy SM, Benjamin IJ, Burke GL, Chait A, Eckel RH, Howard BV, Mitch W, Smith SC, Sowers JR: Diabetes and cardiovascular disease: A statement for healthcare professionals from the American Heart Association. Circulation 100: 1134–1146, 1999
Chatham JC, Forder JR, McNeill JH: The Heart in Diabetes. Norwell, MA: Kluwer Academic Publishers, 1996
Lagadic-Gossmann DL, Buckler KJ, Le Prigent K, Feuvray D: Altered Ca2+ handling in ventricular myocytes isolated from diabetic rats. Am J Physiol 270: H1529–H1537, 1996
Ren J, Davidoff AJ: Diabetes rapidly induces contractile dysfunctions in isolated ventricular myocytes. Am J Physiol 272: H148–H158, 1997
Pierce GN, Russell JC: Regulation of intracellular Ca2+ in the heart during diabetes. Cardiovasc Res 34: 41–47, 1997
Dhalla NS, Pierce GN, Innes IR, Beamish RE: Pathogenesis of cardiac dysfunction in diabetes mellitus. Can J Cardiol 1: 263–281, 1985
Dillmann WH: Diabetes and thyroid hormone-induced changes in cardiac function and their molecular basis. Ann Rev Med 40: 373–394, 1989
Schaffer SW, Ballard-Croft C, Boerth S, Allo SN: Mechanisms underlying depressed Na+/Ca2+ exchanger activity in the diabetic heart. Cardiovasc Res 34: 129–136, 1997
Zhong Y, Ahmed S, Grupp IL, Matlib MA: Altered SR protein expression associated with contractile dysfunction in diabetic rat hearts. Am J Physiol 281: H1137–H1147, 2001
Idris I, Gray S, Donnelly R: Protein kinase C activation: Isozymespecific effects on metabolism and cardiovascular complications in diabetes. Diabetologia 44: 659–673, 2001
Gando S, Hattori Y, Akaishi Y, Nishihira J, Kanno M: Impaired contractile response to beta adrenoceptor stimulation in diabetic rat hearts: Alterations in beta adrenoceptors-G protein-adenylate cyclase system and phospholamban phosphorylation. JPET 282: 475–484, 1997
Shimoni Y, Firek L, Severson D, Giles W: Short-term diabetes alters K+ currents in rat ventricular myocytes. Circ Res 74: 620–628, 1994
Davidoff AJ, Ren J: Low insulin and high glucose induce abnormal relaxation in cultured adult rat ventricular myocytes. Am J Physiol 272: H159–H167, 1997
Ren J, Gintant GA, Miller RE, Davidoff AJ: High extracellular glucose impairs cardiac E-C coupling in a glycosylation-dependent manner.Am J Physiol 273: H2876–H2883, 1997
Dutta K, Carmody MW, Cala SE, Davidoff AJ: Depressed PKA activity contributes to impaired SERCA function and is linked to the pathogenesis of glucose-induced cardiomyopathy. J Mol Cell Cardiol 34: 985–996, 2002
Xiang H, McNeill JH: Protein kinase C activity is altered in diabetic rat hearts. Biochem Biophys Res Commun 187: 703–710, 1992
Malhotra A, Reich D, Reich D, Nakouzi A, Sanghi V, Geenen D, Buttrick P: Experimental diabetes is associated with functional activation of protein kinase Ceand phosphorylation of troponin I in the heart, which are prevented by angiotensin II receptor blockade. Circ Res 81: 1027–1033, 1997
Giles TD, Ouyang J, Kerut EK, Given MB, Allen GE, McIlwain EF, Greenberg SS: Changes in protein kinase C in early cardiomyopathy and in gracilis muscle in the BB/Wor diabetic rat. Am J Physiol 274: H295–H307, 1998
Liu X, Wang J, Takeda N, Binaglia L, Panagia V, Dhalla N: Changes in cardiac protein kinase C activities and isozymes in streptozotocininduced diabetes. Am J Physiol 277: E798–E804, 1999
Mackay K, Mochly-Rosen D: Localization, anchoring, and functions of protein kinase C isozymes in the heart. J Mol Cell Cardiol 33: 1301–1307, 2001
Malhotra A, Kang BPS, Opawumi D, Belizaire W, Meggs LG: Molecular biology of protein kinase C signaling in cardiac myocytes. Mol Cell Biochem 225: 97–107, 2001
Taegtmeyer H, McNulty P, Young M: Adaptation and maladaptation of the heart in diabetes. Part I: General concepts. Circulation 105: 1727–1733, 2002
Kolter T, Uphues I, Eckel J: Molecular analysis of insulin resistance in isolated ventricular cardiomyocytes of obese Zucker rats. Am J Physiol 273: E59–E67, 1997
Koya D, King GL: Protein kinase C activation and the development of diabetic complications. Diabetes 47: 859–866, 1998
Malhotra A, Kang BPS, Cheung S, Opawumi D, Meggs LG: Angiotensin II promotes glucose-induced activation of cardiac protein kinase C isozymes and phosphorylation of Troponin I. Diabetes 50: 1918–1926, 2001
Yasumari K, Kohno M, Kano H, Yokokawa K, Minami M, Yoshikawa J: Mechanisms of action of troglitazone in the prevention of high glucoseinduced migration and proliferation of cultured coronary smooth muscle cells. Circ Res 81: 953–962, 1997
McClain DA, Crook ED: Hexosamines and insulin resistance. Diabetes 45: 1003–1009, 1996
Filippis A, Clark S, Proietto J: Increased flux through the hexosamine bioshynthesis pathway inhibits glucose transport acutely by activation of protein kinase C. Biochem J 324: 981–985, 1997
Eckel J, Reinauer H: Insulin action on glucose transport in isolated cardiac myocytes: Signalling pathways and diabetes-induced alterations. Biochem Soc Trans 18: 1125–1127, 1990
Ren J, Davidoff AJ: a2-Heremans Schmid glycoprotien, a putative inhibitor of tyrosine kinase, prevents glucose toxicity associated with cardiomyocyte dysfunction. Diabetes Metab Res Rev 18: 305–310, 2002
Young ME, McNulty P, Taegtmeyer H: Adaptation and maladaptation of the heart in diabetes: Part II: Potential mechanisms. Circulation 105: 1861–1870, 2002
Tahiliani AG, McNeill JH: Effects of insulin perfusion and altered glucose concentrations on heart function in 3-day and 6-week diabetic rats. Can J Physiol Pharmacol 64: 188–192, 1986
Chatham JC, Gao Z-P, Forder JR: Impact of 1 wk of diabetes on the regulation of myocardial carbohydrate and fatty acid oxidation. Am J Physiol 277: E342–E351, 1999
Depre C, Young ME, Ying J, Ahuja HS, Han Q, Garza N, Davies PJ, Taegtmeyer H: Streptozotocin-induced changes in cardiac gene expression in the absence of severe contractile dysfunction. J Mol Cell Cardiol 32: 985–996, 2000
Belke DD, Betuing S, Tuttle MJ, Graveleau C, Young ME, Pham M, Zhang D, Cooksey RC, McClain DA, Litwin SE, Taegtmeyer H, Severson D, Kahn CR, Abel ED: Insulin signaling coordinately regulates cardiac size, metabolism, and contractile protein isoform expression. J Clin Invest 109: 629–639, 2002
Ren J, Sowers JR, Walsh MF, Brown RA: Reduced contractile response to insulin and IGF-1 in ventricular myocytes from genetically obese Zucker rats. Am J Physiol 279: H1708–H1714, 2000
Eckel J, Wirdeier A, Herberg L, Reinauer H: Insulin resistance in the heart: Studies on isolated cardiocytes of genetically obese Zucker rats. Endocrinology 116: 1529–1534, 1985
Schannwell CM, Schneppenheim M, Perings S, Plehn G, Strauer BE: Left ventricular diastolic dysfunction as an early manifestation of diabetic cardiomyopathy. Cardiology 98: 33–39, 2002
Davidson MB, Ren J, Vlahos CJ, Davidoff AJ: PKCßis elevated in high [glucose]-induced daibetic-like cardiomyopathy. Diabetes 49: A447, 2000
Ren J, Duan J, Hintz KK, Ren BH: High glucose induces cardiac insulinlike growth factor I resistance in ventricular myocytes: Role of Akt and ERK activation. Cardiovasc Res 57: 738–748, 2003
Farese RV: Function and dysfunction of aPKC isoforms for glucose transport in insulin-sensitive and insulin-resistant states. Am J Physiol 283: E1–E11, 2002
Liu LS, Tanaka H, Ishii S, Eckel J: The new antidiabetic drug MCC-555 acutely sensitizes insulin signaling in isolated cardiomyocytes. Endocrinology 139: 4531–4589, 1998
Bähr M, Spelleken M, Bock M, von Holtey M, Kiehn R, Eckel J: Acute and chronic effects of troglitazone (CS-045) on isolated rat ventricular cardiomyocytes. Diabetologia 39: 766–774, 1996
Cooksey RC, Hebert Jr LF, Zhu J-H, Wofford P, Garvey WT, McClain DA: Mechanism of hexosamine-induced insulin resistance in transgenic mice overexpressing glutamine:fructose-6-phosphate amidotransferase: Decreased glucose transporter GLUT4 translocation and reversal by treatment with thiazolidinedione. Endocrinology 140: 1151–1157, 1999
Ren J, Dominguez LJ, Sowers JR, Davidoff AJ: Troglitazone attenuates high glucose-induced abnormalities in relaxation and intracellular calcium in rat ventricular myocytes. Diabetes 45: 1822–1825, 1996
Ren J, Dominguez LJ, Sowers JR, Davidoff AJ: Metformin but not glyburide prevents high glucose-induced abnormalities in relaxation and intracellular Ca2+ transients in adult rat ventricular myocytes. Diabetes 48: 2059–2065, 1999
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Davidoff, A.J., Davidson, M.B., Carmody, M.W. et al. Diabetic cardiomyocyte dysfunction and myocyte insulin resistance: Role of glucose-induced PKC activity. Mol Cell Biochem 262, 155–163 (2004). https://doi.org/10.1023/B:MCBI.0000038231.68078.4b
Issue Date:
DOI: https://doi.org/10.1023/B:MCBI.0000038231.68078.4b