Reversal of isoproterenol-induced downregulation of phospholamban and fkbp12. 6 by cpu0213-mediated antagonism of endothelin receptors

Reversal of isoproterenol-induced downregulation of phospholamban and fkbp12. 6 by cpu0213-mediated antagonism of endothelin receptors

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ABSTRACT AIM: The downregulation of phospholamban (PLB) and FKBP12.6 as a result of β-receptor activation is involved in the pathway(s) of congestive heart failure. We hypothesized that the endothelin (ET)-1 system may link to downregulated PLB and FKBP12.6. METHODS: Rats were subjected to ischemia/reperfusion (I/R) to cause heart failure (HF). 1 mg/kg isoproterenol (ISO) was injected subcutaneously (sc) for 10 d to worsen HF. 30 mg/kg CPU0213 (sc), a dual ET receptor (ETAR/ETBR) antagonist was given from d 6 to d 10. On d 11, cardiac function was assessed together with the determination of mRNA levels of ryanodine receptor 2, calstabin-2 (FKBP12.6), PLB, and sarcoplasmic reticulum Ca2+-ATPase. Isolated adult rat ventricular myocytes were incubated with ISO at 1×10−6 mol/L to set up an _in vitro_ model of HF. Propranolol (PRO), CPU0213, and darusentan (DAR, an ETAR antagonist) were incubated with cardiomyocytes at 1×10−5 mol/L or 1×10−6 mol/L in the presence of ISO (1×10−6 mol/L). Immunocytochemistry and Western blotting were applied for measuring the protein levels of PLB and FKBP12.6. RESULTS: The worsened hemodynamics produced by I/R were exacerbated by ISO pretreatment. The significant downregulation of the gene expression of PLB and FKBP12.6 and worsened cardiac function by ISO were reversed by CPU0213. _In vitro_ ISO 1×10−6 mol/L produced a sharp decline of PLB and FKBP12.6 proteins relative to the control. The downregulation of the protein expression was significantly reversed by the ET receptor antagonist CPU0213 or DAR, comparable to that achieved by PRO. CONCLUSION: This study demonstrates a role of ET in mediating the downregulation of the cardiac Ca2+-handling protein by ISO. SIMILAR CONTENT BEING VIEWED BY OTHERS GLIBENCLAMIDE ALLEVIATES Β ADRENERGIC RECEPTOR ACTIVATION-INDUCED CARDIAC INFLAMMATION Article 04 August 2021 EMPAGLIFLOZIN RESCUES PRO-ARRHYTHMIC AND CA2+ HOMEOSTATIC EFFECTS OF TRANSVERSE AORTIC CONSTRICTION IN INTACT MURINE HEARTS Article Open access 08 July 2024 LUPEOL PROTECTS AGAINST CARDIAC HYPERTROPHY VIA TLR4-PI3K-AKT-NF-ΚB PATHWAYS Article 16 December 2021 ARTICLE PDF REFERENCES * Feldman DS, Carnes CA, Abraham WT, Bristow MR . Mechanisms of disease: beta-adrenergic receptors — alterations in signal transduction and pharmacogenomics in heart failure. _Nat Clin Pract Cardiovasc Med_ 2005; 2: 475–83. Article  CAS  Google Scholar  * Schwinger RH . Treatment of heart failure with beta-receptor-blockers. _Dtsch Med Wochenschr_ 2002; 127: 682–8. Article  CAS  Google Scholar  * Olson EN . A decade of discoveries in cardiac biology. _Nat Med_ 2004; 10: 467–74. Article  CAS  Google Scholar  * Reiken S, Gaburjakova M, Guatimosim S, Gomez AM, D'Armiento J, Burkhoff D, _et al_. Protein kinase A phosphorylation of the cardiac calcium release channel (ryanodine receptor) in normal and failing hearts. Role of phosphatases and response to isoproterenol. _J Biol Chem_ 2003; 278: 444–53. Article  CAS  Google Scholar  * Gupta RC, Mishra S, Rastogi S, Sharov VG, Sabbah HN . Improvement of cardiac sarcoplasmic reticulum calcium cycling in dogs with heart failure following long-term therapy with the Acorn Cardiac Support Device. _Heart Fail Rev_ 2005; 10: 149–55. Article  CAS  Google Scholar  * Yano M, Ikeda Y, Matsuzaki M . Altered intracellular Ca2+ handling in heart failure. _J Clin Invest_ 2005; 115: 556–64. Article  CAS  Google Scholar  * Schmitt JP, Kamisago M, Asahi M, Li GH, Ahmad F, Mende U, _et al_. Dilated cardiomyopathy and heart failure caused by a mutation in phospholamban. _Science_ 2003; 299: 1410–3. Article  CAS  Google Scholar  * Temsah RM, Dyck C, Netticadan T, Chapman D, Elimban V, Dhalla NS . Effect of beta-adrenoceptor blockers on sarcoplasmic reticular function and gene expression in the ischemic-reperfused heart. _J Pharmacol Exp Ther_ 2000; 293: 15–23. CAS  PubMed  Google Scholar  * Wehrens XH, Marks AR . Novel therapeutic approaches for heart failure by normalizing calcium cycling. _Nat Rev Drug Disc_ 2004; 3: 565–73. Article  CAS  Google Scholar  * Ertl G, Bauersachs J . Endothelin receptor antagonists in heart failure: current status and future directions. _Drugs_ 2004; 64: 1029–40. Article  CAS  Google Scholar  * Teerlink JR, McMurray JJ, Bourge RC, Cleland JG, Cotter G, Jondeau G, _et al_. Tezosentan in patients with acute heart failure: design of the Value of Endothelin Receptor Inhibition with Tezosentan in Acute heart failure Study (VERITAS). _Am Heart J_ 2005; 150: 46–53. Article  CAS  Google Scholar  * Anand I, McMurray J, Cohn JN, Konstam MA, Notter T, Quitzau K, _et al_. Long-term effects of darusentan on left-ventricular remodelling and clinical outcomes in the Endothelin A Receptor Antagonist Trial in Heart Failure (EARTH): randomised, double-blind, placebo-controlled trial. _Lancet_ 2004; 364: 347–54. Article  CAS  Google Scholar  * Kelland NF, Webb DJ . Clinical trials of endothelin antagonists in heart failure: a question of dose? _Exp Biol Med (Maywood)_ 2006; 231: 696–9. CAS  Google Scholar  * Qi MY, Xia HJ, Dai DZ, Dai Y . A novel endothelin receptor antagonist CPU0213 improves diabetic cardiac insufficiency attributed to up-regulation of the expression of FKBP12.6, SERCA2a, and PLB in rats. _J Cardiovasc Pharmacol_ 2006; 47: 729–35. Article  CAS  Google Scholar  * Dai DZ, Ji M, Huang M, Liu LG . Endothelin receptor antagonist activity and selective blocking the ETA and ETB of Compound 0213. _J China Pharm Univ_ 2004; 35: 552–7. CAS  Google Scholar  * Li L, Chu Y, Fink GD, Engelhardt JF, Heistad DD, Chen AF . Endothelin-1 stimulates arterial VCAM-1 expression via NADPH oxidase-derived superoxide in mineralocorticoid hypertension. _Hypertension_ 2003; 42: 997–1003. Article  CAS  Google Scholar  * Xu FP, Chen MS, Wang YZ, Yi Q, Lin SB, Chen AF, _et al_. Leptin induces hypertrophy via endothelin-1-reactive oxygen species pathway in cultured neonatal rat cardiomyocytes. _Circulation_ 2004; 110: 1269–75. Article  CAS  Google Scholar  * Zhang TT, Cui B, Dai DZ . Downregulation of Kv4.2 and Kv4.3 channel gene expression in right ventricular hypertrophy induced by monocrotaline in rat. _Acta Pharmacol Sin_ 2004; 25: 226–30. CAS  PubMed  Google Scholar  * Ma YP, Hu HJ, Hao XM, Zhou PA, Wu CH, Dai DZ . Reduced sodium currents in isolated mammalian myocytes treated with chronic L-thyroxine. _Drug Dev Res_ 2003; 58: 111–5. Article  CAS  Google Scholar  * Weil J, Schunkert H . Pathophysiology of chronic heart failure. _Clin Res Cardiol_ 2006; 95 ( Suppl 4): 1–17. Article  CAS  Google Scholar  * Armoundas AA, Rose J, Aggarwal R, Stuyvers BD, O'rourke B, Kass DA, _et al_. Cellular and molecular determinants of altered Ca2+ handling in the failing rabbit heart: primary defects in SR Ca2+ uptake and release mechanisms. _Am J Physiol Heart Circ Physiol_ 2007; 292: H1607–18. Article  CAS  Google Scholar  * Sucharov CC . Beta-adrenergic pathways in human heart failure. _Expert Rev Cardiovasc Ther_ 2007; 5: 119–24. Article  CAS  Google Scholar  * Armoundas AA, Rose J, Aggarwal R, Stuyvers B, O'Rourke B, Kass DA, _et al_. Cellular and molecular determinants of altered Ca2+ handling in the failing rabbit heart: Primary defects in SR Ca2+ uptake and release mechanisms. _Am J Physiol Heart Circ Physiol_ 2007; 292: H1607–18. Article  CAS  Google Scholar  * Saliaris AP, Amado LC, Minhas KM, Schuleri KH, Lehrke S, StJohn, M, _et al_. Chronic allopurinol administration ameliorates maladaptive alterations in Ca2+ cycling proteins and beta-adrenergic hyporesponsiveness in heart failure. _Am J Physiol Heart Circ Physiol_ 2007; 292: H1328–35. Article  CAS  Google Scholar  * Huang F, Shan J, Reiken S, Wehrens XH, Marks AR . Analysis of calstabin2 (FKBP12.6)-ryanodine receptor interactions: rescue of heart failure by calstabin2 in mice. _Proc Natl Acad Sci USA_ 2006; 103: 3456–61. Article  CAS  Google Scholar  * Phrommintikul A, Chattipakorn N . Roles of cardiac ryanodine receptor in heart failure and sudden cardiac death. _Int J Cardiol_ 2006; 112: 142–52. Article  Google Scholar  * Marin-Garcia J, Goldenthal MJ, Moe GW . Selective endothelin receptor blockade reverses mitochondrial dysfunction in canine heart failure. _J Card Fail_ 2002; 8: 326–32. Article  CAS  Google Scholar  * Xia HJ, Dai DZ, Dai Y . Up-regulated inflammatory factors endothelin, NFκB, TNF-α and iNOS involved in exaggerated cardiac arrhythmias in L-thyroxine-induced cardiomyopathy are suppressed by darusentan in rats. _Life Sci_ 2006; 79: 1812–9. Article  CAS  Google Scholar  Download references AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Research Division of Pharmacology, China Pharmaceutical University, Nanjing, 210009, China Yu Feng, Xiao-yun Tang, De-zai Dai & Yin Dai Authors * Yu Feng View author publications You can also search for this author inPubMed Google Scholar * Xiao-yun Tang View author publications You can also search for this author inPubMed Google Scholar * De-zai Dai View author publications You can also search for this author inPubMed Google Scholar * Yin Dai View author publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to De-zai Dai. ADDITIONAL INFORMATION Project supported by the National Natural Science Foundation of China (No 30670760). RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Feng, Y., Tang, Xy., Dai, Dz. _et al._ Reversal of isoproterenol-induced downregulation of phospholamban and FKBP12.6 by CPU0213-mediated antagonism of endothelin receptors. _Acta Pharmacol Sin_ 28, 1746–1754 (2007). https://doi.org/10.1111/j.1745-7254.2007.00650.x Download citation * Received: 04 February 2007 * Accepted: 11 May 2007 * Issue Date: 01 November 2007 * DOI: https://doi.org/10.1111/j.1745-7254.2007.00650.x SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content: Get shareable link Sorry, a shareable link is not currently available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative KEYWORDS * heart failure * endothelin receptor antagonists * phospholamban * FKBP12.6 * propranolol * CPU0213

ABSTRACT AIM: The downregulation of phospholamban (PLB) and FKBP12.6 as a result of β-receptor activation is involved in the pathway(s) of congestive heart failure. We hypothesized that the


endothelin (ET)-1 system may link to downregulated PLB and FKBP12.6. METHODS: Rats were subjected to ischemia/reperfusion (I/R) to cause heart failure (HF). 1 mg/kg isoproterenol (ISO) was


injected subcutaneously (sc) for 10 d to worsen HF. 30 mg/kg CPU0213 (sc), a dual ET receptor (ETAR/ETBR) antagonist was given from d 6 to d 10. On d 11, cardiac function was assessed


together with the determination of mRNA levels of ryanodine receptor 2, calstabin-2 (FKBP12.6), PLB, and sarcoplasmic reticulum Ca2+-ATPase. Isolated adult rat ventricular myocytes were


incubated with ISO at 1×10−6 mol/L to set up an _in vitro_ model of HF. Propranolol (PRO), CPU0213, and darusentan (DAR, an ETAR antagonist) were incubated with cardiomyocytes at 1×10−5


mol/L or 1×10−6 mol/L in the presence of ISO (1×10−6 mol/L). Immunocytochemistry and Western blotting were applied for measuring the protein levels of PLB and FKBP12.6. RESULTS: The worsened


hemodynamics produced by I/R were exacerbated by ISO pretreatment. The significant downregulation of the gene expression of PLB and FKBP12.6 and worsened cardiac function by ISO were


reversed by CPU0213. _In vitro_ ISO 1×10−6 mol/L produced a sharp decline of PLB and FKBP12.6 proteins relative to the control. The downregulation of the protein expression was significantly


reversed by the ET receptor antagonist CPU0213 or DAR, comparable to that achieved by PRO. CONCLUSION: This study demonstrates a role of ET in mediating the downregulation of the cardiac


Ca2+-handling protein by ISO. SIMILAR CONTENT BEING VIEWED BY OTHERS GLIBENCLAMIDE ALLEVIATES Β ADRENERGIC RECEPTOR ACTIVATION-INDUCED CARDIAC INFLAMMATION Article 04 August 2021


EMPAGLIFLOZIN RESCUES PRO-ARRHYTHMIC AND CA2+ HOMEOSTATIC EFFECTS OF TRANSVERSE AORTIC CONSTRICTION IN INTACT MURINE HEARTS Article Open access 08 July 2024 LUPEOL PROTECTS AGAINST CARDIAC


HYPERTROPHY VIA TLR4-PI3K-AKT-NF-ΚB PATHWAYS Article 16 December 2021 ARTICLE PDF REFERENCES * Feldman DS, Carnes CA, Abraham WT, Bristow MR . Mechanisms of disease: beta-adrenergic


receptors — alterations in signal transduction and pharmacogenomics in heart failure. _Nat Clin Pract Cardiovasc Med_ 2005; 2: 475–83. Article  CAS  Google Scholar  * Schwinger RH .


Treatment of heart failure with beta-receptor-blockers. _Dtsch Med Wochenschr_ 2002; 127: 682–8. Article  CAS  Google Scholar  * Olson EN . A decade of discoveries in cardiac biology. _Nat


Med_ 2004; 10: 467–74. Article  CAS  Google Scholar  * Reiken S, Gaburjakova M, Guatimosim S, Gomez AM, D'Armiento J, Burkhoff D, _et al_. Protein kinase A phosphorylation of the


cardiac calcium release channel (ryanodine receptor) in normal and failing hearts. Role of phosphatases and response to isoproterenol. _J Biol Chem_ 2003; 278: 444–53. Article  CAS  Google


Scholar  * Gupta RC, Mishra S, Rastogi S, Sharov VG, Sabbah HN . Improvement of cardiac sarcoplasmic reticulum calcium cycling in dogs with heart failure following long-term therapy with the


Acorn Cardiac Support Device. _Heart Fail Rev_ 2005; 10: 149–55. Article  CAS  Google Scholar  * Yano M, Ikeda Y, Matsuzaki M . Altered intracellular Ca2+ handling in heart failure. _J Clin


Invest_ 2005; 115: 556–64. Article  CAS  Google Scholar  * Schmitt JP, Kamisago M, Asahi M, Li GH, Ahmad F, Mende U, _et al_. Dilated cardiomyopathy and heart failure caused by a mutation


in phospholamban. _Science_ 2003; 299: 1410–3. Article  CAS  Google Scholar  * Temsah RM, Dyck C, Netticadan T, Chapman D, Elimban V, Dhalla NS . Effect of beta-adrenoceptor blockers on


sarcoplasmic reticular function and gene expression in the ischemic-reperfused heart. _J Pharmacol Exp Ther_ 2000; 293: 15–23. CAS  PubMed  Google Scholar  * Wehrens XH, Marks AR . Novel


therapeutic approaches for heart failure by normalizing calcium cycling. _Nat Rev Drug Disc_ 2004; 3: 565–73. Article  CAS  Google Scholar  * Ertl G, Bauersachs J . Endothelin receptor


antagonists in heart failure: current status and future directions. _Drugs_ 2004; 64: 1029–40. Article  CAS  Google Scholar  * Teerlink JR, McMurray JJ, Bourge RC, Cleland JG, Cotter G,


Jondeau G, _et al_. Tezosentan in patients with acute heart failure: design of the Value of Endothelin Receptor Inhibition with Tezosentan in Acute heart failure Study (VERITAS). _Am Heart


J_ 2005; 150: 46–53. Article  CAS  Google Scholar  * Anand I, McMurray J, Cohn JN, Konstam MA, Notter T, Quitzau K, _et al_. Long-term effects of darusentan on left-ventricular remodelling


and clinical outcomes in the Endothelin A Receptor Antagonist Trial in Heart Failure (EARTH): randomised, double-blind, placebo-controlled trial. _Lancet_ 2004; 364: 347–54. Article  CAS 


Google Scholar  * Kelland NF, Webb DJ . Clinical trials of endothelin antagonists in heart failure: a question of dose? _Exp Biol Med (Maywood)_ 2006; 231: 696–9. CAS  Google Scholar  * Qi


MY, Xia HJ, Dai DZ, Dai Y . A novel endothelin receptor antagonist CPU0213 improves diabetic cardiac insufficiency attributed to up-regulation of the expression of FKBP12.6, SERCA2a, and PLB


in rats. _J Cardiovasc Pharmacol_ 2006; 47: 729–35. Article  CAS  Google Scholar  * Dai DZ, Ji M, Huang M, Liu LG . Endothelin receptor antagonist activity and selective blocking the ETA


and ETB of Compound 0213. _J China Pharm Univ_ 2004; 35: 552–7. CAS  Google Scholar  * Li L, Chu Y, Fink GD, Engelhardt JF, Heistad DD, Chen AF . Endothelin-1 stimulates arterial VCAM-1


expression via NADPH oxidase-derived superoxide in mineralocorticoid hypertension. _Hypertension_ 2003; 42: 997–1003. Article  CAS  Google Scholar  * Xu FP, Chen MS, Wang YZ, Yi Q, Lin SB,


Chen AF, _et al_. Leptin induces hypertrophy via endothelin-1-reactive oxygen species pathway in cultured neonatal rat cardiomyocytes. _Circulation_ 2004; 110: 1269–75. Article  CAS  Google


Scholar  * Zhang TT, Cui B, Dai DZ . Downregulation of Kv4.2 and Kv4.3 channel gene expression in right ventricular hypertrophy induced by monocrotaline in rat. _Acta Pharmacol Sin_ 2004;


25: 226–30. CAS  PubMed  Google Scholar  * Ma YP, Hu HJ, Hao XM, Zhou PA, Wu CH, Dai DZ . Reduced sodium currents in isolated mammalian myocytes treated with chronic L-thyroxine. _Drug Dev


Res_ 2003; 58: 111–5. Article  CAS  Google Scholar  * Weil J, Schunkert H . Pathophysiology of chronic heart failure. _Clin Res Cardiol_ 2006; 95 ( Suppl 4): 1–17. Article  CAS  Google


Scholar  * Armoundas AA, Rose J, Aggarwal R, Stuyvers BD, O'rourke B, Kass DA, _et al_. Cellular and molecular determinants of altered Ca2+ handling in the failing rabbit heart: primary


defects in SR Ca2+ uptake and release mechanisms. _Am J Physiol Heart Circ Physiol_ 2007; 292: H1607–18. Article  CAS  Google Scholar  * Sucharov CC . Beta-adrenergic pathways in human


heart failure. _Expert Rev Cardiovasc Ther_ 2007; 5: 119–24. Article  CAS  Google Scholar  * Armoundas AA, Rose J, Aggarwal R, Stuyvers B, O'Rourke B, Kass DA, _et al_. Cellular and


molecular determinants of altered Ca2+ handling in the failing rabbit heart: Primary defects in SR Ca2+ uptake and release mechanisms. _Am J Physiol Heart Circ Physiol_ 2007; 292: H1607–18.


Article  CAS  Google Scholar  * Saliaris AP, Amado LC, Minhas KM, Schuleri KH, Lehrke S, StJohn, M, _et al_. Chronic allopurinol administration ameliorates maladaptive alterations in Ca2+


cycling proteins and beta-adrenergic hyporesponsiveness in heart failure. _Am J Physiol Heart Circ Physiol_ 2007; 292: H1328–35. Article  CAS  Google Scholar  * Huang F, Shan J, Reiken S,


Wehrens XH, Marks AR . Analysis of calstabin2 (FKBP12.6)-ryanodine receptor interactions: rescue of heart failure by calstabin2 in mice. _Proc Natl Acad Sci USA_ 2006; 103: 3456–61. Article


  CAS  Google Scholar  * Phrommintikul A, Chattipakorn N . Roles of cardiac ryanodine receptor in heart failure and sudden cardiac death. _Int J Cardiol_ 2006; 112: 142–52. Article  Google


Scholar  * Marin-Garcia J, Goldenthal MJ, Moe GW . Selective endothelin receptor blockade reverses mitochondrial dysfunction in canine heart failure. _J Card Fail_ 2002; 8: 326–32. Article 


CAS  Google Scholar  * Xia HJ, Dai DZ, Dai Y . Up-regulated inflammatory factors endothelin, NFκB, TNF-α and iNOS involved in exaggerated cardiac arrhythmias in L-thyroxine-induced


cardiomyopathy are suppressed by darusentan in rats. _Life Sci_ 2006; 79: 1812–9. Article  CAS  Google Scholar  Download references AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Research


Division of Pharmacology, China Pharmaceutical University, Nanjing, 210009, China Yu Feng, Xiao-yun Tang, De-zai Dai & Yin Dai Authors * Yu Feng View author publications You can also


search for this author inPubMed Google Scholar * Xiao-yun Tang View author publications You can also search for this author inPubMed Google Scholar * De-zai Dai View author publications You


can also search for this author inPubMed Google Scholar * Yin Dai View author publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to


De-zai Dai. ADDITIONAL INFORMATION Project supported by the National Natural Science Foundation of China (No 30670760). RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE


CITE THIS ARTICLE Feng, Y., Tang, Xy., Dai, Dz. _et al._ Reversal of isoproterenol-induced downregulation of phospholamban and FKBP12.6 by CPU0213-mediated antagonism of endothelin


receptors. _Acta Pharmacol Sin_ 28, 1746–1754 (2007). https://doi.org/10.1111/j.1745-7254.2007.00650.x Download citation * Received: 04 February 2007 * Accepted: 11 May 2007 * Issue Date: 01


November 2007 * DOI: https://doi.org/10.1111/j.1745-7254.2007.00650.x SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content: Get shareable link


Sorry, a shareable link is not currently available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative KEYWORDS * heart failure *


endothelin receptor antagonists * phospholamban * FKBP12.6 * propranolol * CPU0213