nach oben

Cardiovascular Drugs and Therapy

Erschienen in:

01.10.2013 | ORIGINAL ARTICLE

The Consequences of Long-Term Glycogen Synthase Kinase-3 Inhibition on Normal and Insulin Resistant Rat Hearts

verfasst von: T. B. Flepisi, Amanda Lochner, Barbara Huisamen

Erschienen in: Cardiovascular Drugs and Therapy | Ausgabe 5/2013

Einloggen, um Zugang zu erhalten

Abstract

Background

Glycogen synthase kinase-3 (GSK-3) is a serine-threonine protein kinase, discovered as a regulator of glycogen synthase. GSK-3 may regulate the expression of SERCA-2a potentially affecting myocardial contractility. It is known to phosphorylate and inhibit IRS-1, thus disrupting insulin signalling. This study aimed to determine whether myocardial GSK-3 protein and its substrate proteins are dysregulated in obesity and insulin resistance, and whether chronic GSK-3 inhibition can prevent or reverse this.

Methods

Weight matched male Wistar rats were rendered obese by hyperphagia using a special diet (DIO) for 16 weeks and compared to chow fed controls. Half of each group was treated with the GSK-3 inhibitor CHIR118637 (30 mg/kg/day) from week 12 to16 of the diet period. Biometric and biochemical parameters were measured and protein expression determined by Western blotting and specific antibodies. Ca²⁺ATPase activity was determined spectrophotometrically. Cardiomyocytes were prepared by collagenase perfusion and insulin stimulated 2-deoxy-glucose uptake determined.

Results

DIO rats were significantly heavier than controls, associated with increased intra-peritoneal fat and insulin resistance. GSK-3 inhibition did not affect weight but improved insulin resistance, also on cellular level. It had no effect on GSK-3 expression but elevated its phospho/total ratio and elevated IRS-2 expression. Obesity lowered SERCA-2a expression and activity while GSK-3 inhibition alleviated this. The phospho/total ratio of phospholamban underscored inhibition of SERCA-2a in obesity. In addition, signs of myocardial hypertrophy were observed in treated control rats.

Conclusion

GSK-3 inhibition could not reverse all the detrimental effects of obesity but may be harmful in normal rat hearts. It regulates IRS-2, SERCA-2a and phospholamban expression but not IRS-1.

Hemmings BA, Yellowlees D, Kernohan JC, Cohen P. Purification of glycogen synthase kinase 3 from rabbit skeletal muscle. Co-purification with the activation factor(FA) of the (Mg-ATP) dependent protein phosphatase. Eur J Biochem. 1981;119:443–51.PubMedCrossRef

Embi N, Rylatt DB, Cohen P. Glycogen synthase kinase-3 from rabbit skeletal muscle. Separation from cyclic-AMP-dependent protein kinase and phosphorylase kinase. Eur J Biochem. 1980;107:519–27.PubMedCrossRef

Huisamen B, Lochner A. GSK-3 protein and the heart: friend or foe? SAHeart. 2010;7:48–57.

Woodgett JR. Judging a protein by more than its name: GSK-3. Sci STKE. 2001;2001(100):re12.PubMedCrossRef

Ciaraldi TP, Nikoulina SE, Bandukwala RA, Carter L, Henry RR. Role of glycogen synthase kinase-3 alpha in insulin action in cultured human skeletal muscle cells. Endocrinology. 2007;148:4393–9.PubMedCrossRef

Woodgett JR. Molecular cloning and expression of glycogen synthase kinase-3/factor A. EMBO J. 1990;9:2431–8.PubMed

Doble BW, Woodgett JR. GSK-3: tricks of the trade for a multi-tasking kinase. J Cell Sci. 2003;116:1175–86.PubMedCrossRef

Parker PJ, Caudwell FB, Cohen P. Glycogen synthase from rabbit skeletal muscle; effect of insulin on the state of phosphorylation of the seven phosphoserine residues in vivo. Eur J Biochem. 1983;130:227–34.PubMedCrossRef

Mora A, Sakamoto K, McManus EJ, Alessi DR. Role of the PDK-1-PKB-GSK-3 pathway in regulating glycogen synthase and glucose uptake in the heart. FEBS Lett. 2005;579:3632–8.PubMedCrossRef

10.

Eldar-Finkelman H, Argast GM, Foord O, Fischer EH, Krebs EG. Expression and characterization of glycogen synthase kinase-3 mutants and their effect on glycogen synthase activity in intact cells. Proc Natl Acad Sci U S A. 1996;93(19):10228–33.PubMedCrossRef

11.

Michael A, Haq S, Chen X, Hsich E, Cui L, Walters B, et al. Glycogen synthase kinase-3beta regulates growth, calcium homeostasis, and diastolic function in the heart. J Biol Chem. 2004;279:21383–93.PubMedCrossRef

12.

Omar MA, Wang L, Clanachan AS. Cardioprotection by GSK-3 inhibition: role of enhanced glycogen synthesis and attenuation of calcium overload. Cardiovasc Res. 2010;86(3):478–86.PubMedCrossRef

13.

Takeda N. Cardiac disturbances in diabetes mellitus. Pathophysiology. 2010;17:83–8.PubMedCrossRef

14.

Asahi M, Nakayama H, Tada M, Otsu K. Regulation of sarco(endo)plasmic reticulum Ca2+ adenosine triphosphatase by phospholamban and sarcolipin: implication for cardiac hypertrophy and failure. Trends Cardiovasc Med. 2003;13:152–7.PubMedCrossRef

15.

Huisamen B, Dietrich D, Blackhurst D, Flepisi B, Lochner A. Early cardiovascular changes occurring in diet-induced obese, insulin resistant rats. Mol Cell Biochem. 2012;368:37–45.PubMedCrossRef

16.

Huisamen B, Pêrel SJC, Friedrich SO, Salie R, Strijdom H, Lochner A. AngII receptor antagonism improves nitric oxide production, eNOS and PKB expression in hearts from a rat model of insulin resistance. Mol Cell Biochem. 2011;349:21–31.PubMedCrossRef

17.

Huisamen B, Genis A, Lochner A. Pre-treatment with a DPP-4 inhibitor is infarct sparing in hearts from obese, pre-diabetic rats. Cardiovasc Drugs Ther. 2011;25:13–20.PubMedCrossRef

18.

Henriksen EJ, Dokken BB. Role of glycogen synthase kinase-3 in insulin resistance and type 2 diabetes. Curr Drug Targets. 2006;7:1435–41.PubMedCrossRef

19.

Cline GW, Johnson K, Regittnig W, Perret P, Tozzo E, Xiao L, et al. Effects of a novel glycogen synthase kinase-3 inhibitor on insulin-stimulated glucose metabolism in Zucker diabetic fatty (fa/fa) rats. Diabetes. 2002;51:2903–10.PubMedCrossRef

20.

Meijer L, Flajolet M, Greengard P. Pharmacological inhibitors of glycogen synthase kinase 3. Trends Pharmacol Sci. 2004;25:471–80.PubMedCrossRef

21.

Wagman AS, Johnson KW, Bussiere DE. Discovery and development of GSK3 inhibitors for the treatment of type 2 diabetes. Curr Pharm Des. 2004;10:1105–37.PubMedCrossRef

22.

Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248–54.PubMedCrossRef

23.

Feher JJ, Davis MD. Isolation of rat cardiac sarcoplasmic reticulum with improved Ca²⁺ uptake and ryanodine binding. J Mol Cell Cardiol. 1991;23:249–58.PubMedCrossRef

24.

Bers DM. Isolation and characterization of cardiac sarcolemma. Biochem Biophys Acta. 1979;555:131–6.PubMedCrossRef

25.

Fiske CH, Subbarow Y. The colorimetric determination of phosphorus. J Biol Chem. 1925;66:375–400.

26.

Donthi R, Huisamen B, Lochner A. The effect of vanadate and insulin on glucose transport in isolated adult rat cardiomyocytes. Cardiovasc Drugs Ther. 2000;14:463–70.PubMedCrossRef

27.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193:265–75.PubMed

28.

Suarez J, Scott B, Dillmann WH. Conditional increase in SERCA2a protein is able to reverse contractile dysfunction and abnormal calcium flux in established diabetic cardiomyopathy. Am J Physiol Regul Integr Comp Physiol. 2008;295:R1439–45.PubMedCrossRef

29.

Eldar-Finkelman H, Licht-Murava A, Pietrokovski S, Eisenstein M. Substrate competitive GSK-3 inhibitors—strategy and implications. Biochim Biophys Acta. 1804;2010:598–603.

30.

Martinez A. Preclinical efficacy on GSK-3 inhibitors: towards a future generation of powerful drugs. Med Res Rev. 2008;28:773–96.PubMedCrossRef

31.

Lee S, Yang WK, Song JH, Ra YM, Jeong JH, Choe W, et al. Anti-obesity effects of 3-hydroxychromone derivative, a novel small-molecule inhibitor of glycogen synthase kinase-3. Biochem Pharmacol. 2013;85:965–76.PubMedCrossRef

32.

Abel ED, Litwin SE, Sweeney G. Cardiac remodeling in obesity. Physiol Rev. 2008;88:389–419.PubMedCrossRef

33.

Chess DJ, Stanley WC. Role of diet and fuel overabundance in the development and progression of heart failure. Cardiovasc Res. 2008;79:269–78.PubMedCrossRef

34.

Wolk R. Arrhythmogenic mechanisms in left ventricular hypertrophy. Europace. 2000;2:216–23.PubMedCrossRef

35.

Haq S, Choukroun G, Kang ZB, Ranu H, Matsui T, Rosenzweig A, et al. Glycogen synthase kinase-3beta is a negative regulator of cardiomyocyte hypertrophy. J Cell Biol. 2000;151:117–30.PubMedCrossRef

36.

Roach PJ. Control of glycogen synthase by hierarchal protein phosphorylation. FASEB J. 1990;4:2961–8.PubMed

37.

Zhang W, DePaoli-Roach AA, Roach PJ. Mechanisms of multisite phosphorylation and inactivation of rabbit muscle glycogen synthase. Arch Biochem Biophys. 1993;304:219–25.PubMedCrossRef

38.

Shulman GI, Rothman DL, Jue T, Stein P, DeFronzo RA, Shulman RG. Quantitation of muscle glycogen synthesis in normal subjects and subjects with non-insulin-dependent diabetes by 13C nuclear magnetic resonance spectroscopy. N Engl J Med. 1990;322:223–8.PubMedCrossRef

39.

Cline GW, Rothman DL, Magnusson I, Katz LD, Shulman GI. 13C-nuclear magnetic resonance spectroscopy studies of hepatic glucose metabolism in normal subjects and subjects with insulin-dependent diabetes mellitus. J Clin Invest. 1994;94:2369–76.PubMedCrossRef

40.

Ring DB, Johnson KW, Henriksen EJ, Nuss JM, Goff D, Kinnick TR, et al. Selective glycogen synthase kinase 3 inhibitors potentiate insulin activation of glucose transport and utilization in vitro and in vivo. Diabetes. 2003;52:588–95.PubMedCrossRef

41.

Coghlan MP, Culbert AA, Cross DA, Corcoran SL, Yates JW, Pearce NJ, et al. Selective small molecule inhibitors of glycogen synthase kinase-3 modulate glycogen metabolism and gene transcription. Chem Biol. 2000;7:793–803.PubMedCrossRef

42.

Nikoulina SE, Ciaraldi TP, Mudaliar S, Carter L, Johnson K, Henry RR. Inhibition of glycogen synthase kinase 3 improves insulin action and glucose metabolism in human skeletal muscle. Diabetes. 2002;51:2190–8.PubMedCrossRef

43.

Gao HK, Yin Z, Zhou N, Feng XY, Gao F, Wang HC. Glycogen synthase kinase 3 inhibition protects the heart from acute ischemia-reperfusion injury via inhibition of inflammation and apoptosis. J Cardiovasc Pharmacol. 2008;52:286–92.PubMedCrossRef

44.

MacAulay K, Blair AS, Hajduch E, Terashima T, Baba O, Sutherland C, et al. Constitutive activation of GSK3 down-regulates glycogen synthase abundance and glycogen deposition in rat skeletal muscle cells. J Biol Chem. 2005;280:9509–18.PubMedCrossRef

45.

Dihlmann S, Kloor M, Fallsehr C, Von Knebel DM. Regulation of Akt1 expression by beta-catenin/Tcf/Lef signalling in colorectal cancer cells. Carcinogenesis. 2005;26:1503–12.PubMedCrossRef

46.

Henriksen EJ, Teachey MK. Short-term in vitro inhibition of glycogen synthase kinase 3 potentiates insulin signalling in type I skeletal muscle of Zucker diabetic Fatty rats. Metabolism. 2007;56:931–8.PubMedCrossRef

47.

Wade A. GSK-3 inhibitors and insulin receptor signalling in health, disease and therapeutics. Front Biosci. 2009;14:1558–70.CrossRef

48.

Ciaraldi TP, Carter L, Mudaliar S, Henry RR. GSK-3beta and control of glucose metabolism and insulin action in human skeletal muscle. Mol Cell Endocrinol. 2010;315:153–8.PubMedCrossRef

49.

Araki E, Lipes MA, Patti ME, Brüning JC, Haag 3rd B, Johnson RS, et al. Alternative pathway of insulin signalling in mice with targeted disruption of the IRS-1 gene. Nature. 1994;372:186–90.PubMedCrossRef

50.

Vittorini S, Storti S, Parri MS, Cerillo AG, Clerico A. SERCA2a, phospholamban, sarcolipin, and ryanodine receptors gene expression in children with congenital heart defects. Mol Med. 2007;13:105–11.PubMedCrossRef

51.

James P, Inui M, Tada M, Chiesi M, Carafoli E. Nature and site of phospholamban regulation of the Ca2+ pump of sarcoplasmic reticulum. Nature. 1989;342:90–2.PubMedCrossRef

Titel: The Consequences of Long-Term Glycogen Synthase Kinase-3 Inhibition on Normal and Insulin Resistant Rat Hearts
verfasst von: T. B. Flepisi
Amanda Lochner
Barbara Huisamen
Publikationsdatum: 01.10.2013
Verlag: Springer US
Erschienen in: Cardiovascular Drugs and Therapy / Ausgabe 5/2013
Print ISSN: 0920-3206
Elektronische ISSN: 1573-7241
DOI: https://doi.org/10.1007/s10557-013-6467-8

Neu im Fachgebiet Kardiologie

23.04.2024 | Ablationstherapie | Nachrichten

Update Kardiologie

Bestellen Sie unseren Fach-Newsletter und bleiben Sie gut informiert.

Newsletter bestellen

Live-Webinar: Aktuelle Leitlinien bei Herz-Kreislauf-Erkrankungen

Springer Medizin

The Consequences of Long-Term Glycogen Synthase Kinase-3 Inhibition on Normal and Insulin Resistant Rat Hearts

Abstract

Background

Methods

Results

Conclusion

Neu im Fachgebiet Kardiologie

Wie erfolgreich ist eine Re-Ablation nach Rezidiv?

Europäische Ernährungsgewohnheiten: Das sind die häufigsten Fehler

Parodontalbehandlung verbessert Prognose bei Katheterablation

Antihypertensiva schützen auch alte Menschen noch vor Demenz

Update Kardiologie

Live-Webinar: Aktuelle Leitlinien bei Herz-Kreislauf-Erkrankungen

Springer Medizin

Abstract

Background

Methods

Results

Conclusion

Bitte loggen Sie sich ein, um Zugang zu diesem Inhalt zu erhalten

Weitere Artikel der Ausgabe 5/2013

Erratum to: Comparison of Clinical Efficacy and Safety of Clopidogrel Resinate With Clopidogrel Bisulfate in Patients Undergoing Percutaneous Coronary Intervention

Comparison of Clinical Efficacy and Safety of Clopidogrel Resinate With Clopidogrel Bisulfate in Patients Undergoing Percutaneous Coronary Intervention

Administration by Gavage is the Rule

Long Term Effects of Epoetin Alfa in Patients with ST- Elevation Myocardial Infarction

Statins as a New Therapeutic Perspective in Myocarditis and Postmyocarditis Dilated Cardiomyopathy

Pitavastatin Regulates Helper T-Cell Differentiation and Ameliorates Autoimmune Myocarditis in Mice

Neu im Fachgebiet Kardiologie

Wie erfolgreich ist eine Re-Ablation nach Rezidiv?

Europäische Ernährungsgewohnheiten: Das sind die häufigsten Fehler

Parodontalbehandlung verbessert Prognose bei Katheterablation

Antihypertensiva schützen auch alte Menschen noch vor Demenz

Update Kardiologie