Skip to main content

Advertisement

SpringerLink
Log in

A Gene Expression Profile of the Myocardial Response to Clenbuterol

  • Published:
Journal of Cardiovascular Translational Research Aims and scope Submit manuscript

Abstract

Clenbuterol is currently being used as part of a clinical trial into a novel therapeutic approach for the treatment of end-stage heart failure. The purpose of this study was to determine the global pattern of myocardial gene expression in response to clenbuterol and to identify novel targets and pathways involved. Rats were treated with clenbuterol (n = 6) or saline (n = 6) for periods of 1, 3, 9, or 28 days. Rats treated for 28 days were also subject to continuous electrocardiogram analysis using implantable telemetry. RNA was extracted from rats at days 1 and 28 and used from microarray analysis, and further samples from rats at days 1, 3, 9, and 28 were used for analysis by real-time polymerase chain reaction. Clenbuterol treatment induced rapid development of cardiac hypertrophy with increased muscle mass at day 1 and elevated heart rate and QT interval throughout the 28-day period. Microarray analysis revealed a marked but largely transitory change in gene expression with 1,423 genes up-regulated and 964 genes down-regulated at day 1. Up-regulated genes revealed an unexpected association with angiogenesis and integrin-mediated cell adhesion and signaling. Moreover, direct treatment of endothelial cells cultured in vitro resulted in increased cell proliferation and tube formation. Our data show that clenbuterol treatment is associated with rapid cardiac hypertrophy and identify angiogenesis and integrin signaling as novel pathways of clenbuterol action. The data have implications both for our understanding of the physiologic hypertrophy induced by clenbuterol and for treatment of heart failure.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Jessup, M., & Brozena, S. (2003). Heart failure. New England Journal of Medicine, 348(20), 2007–2018.

    Article  PubMed  Google Scholar 

  2. Frazier, O. H., & Myers, T. J. (1999). Left ventricular assist system as a bridge to myocardial recovery. Annals of Thoracic Surgery, 68(2), 734–741.

    Article  PubMed  CAS  Google Scholar 

  3. Maybaum, S., Mancini, D., Xydas, S., Starling, R. C., Aaronson, K., Pagani, F. D., et al. (2007). Cardiac improvement during mechanical circulatory support: a prospective multicenter study of the LVAD Working Group. Circulation, 115(19), 2497–2505.

    Article  PubMed  Google Scholar 

  4. Birks, E. J., Tansley, P. D., Hardy, J., George, R. S., Bowles, C. T., Burke, M., et al. (2006). Left ventricular assist device and drug therapy for the reversal of heart failure. New England Journal of Medicine, 355(18), 1873–1884.

    Article  PubMed  CAS  Google Scholar 

  5. Yacoub, M. H. (2001). A novel strategy to maximize the efficacy of left ventricular assist devices as a bridge to recovery. European Heart Journal, 22(7), 534–540.

    Article  PubMed  CAS  Google Scholar 

  6. Salorinne, Y., Stenius, B., Tukiainen, P., & Poppius, H. (1975). Double-blind cross-over comparison of clenbuterol and salbutamol tablets in asthmatic out-patients. European Journal of Clinical Pharmacology, 8(3–4), 189–195.

    Article  PubMed  CAS  Google Scholar 

  7. Rockman, H. A., Koch, W. J., & Lefkowitz, R. J. (2002). Seven-transmembrane-spanning receptors and heart function. Nature, 415(6868), 206–212.

    Article  PubMed  CAS  Google Scholar 

  8. Wong, K., Boheler, K. R., Petrou, M., & Yacoub, M. H. (1997). Pharmacological modulation of pressure-overload cardiac hypertrophy: Changes in ventricular function, extracellular matrix and gene expression. Circulation, 96, 2239–2246.

    PubMed  CAS  Google Scholar 

  9. Wong, K., Boheler, K. R., Bishop, J., Petrou, M., & Yacoub, M. (1998). Clenbuterol induces cardiac hypertrophy with normal function, morphological and molecular features. Cardiovascular Research, 37, 115–122.

    Article  PubMed  CAS  Google Scholar 

  10. Petrou, M., Wynne, D. G., Boheler, K. R., & Yacoub, M. H. (1995). Clenbuterol induces hypertrophy of the latissimus dorsi muscle and heart in the rat with molecular and phenotypic changes. Circulation, 92(suppl II), 483–489.

    CAS  Google Scholar 

  11. Soppa, G. K., Smolenski, R. T., Latif, N., Yuen, A. H., Malik, A., Karbowska, J., et al. (2005). Effects of chronic administration of clenbuterol on function and metabolism of adult rat cardiac muscle. American Journal of Physiology. Heart and Circulatory Physiology, 288(3), H1468–H1476.

    Article  PubMed  CAS  Google Scholar 

  12. Soppa, G. K., Lee, J., Stagg, M. A., Felkin, L. E., Barton, P. J., Siedlecka, U., et al. (2008). Role and possible mechanisms of clenbuterol in enhancing reverse remodelling during mechanical unloading in murine heart failure. Cardiovascular Research, 77(4), 695–706.

    Article  PubMed  CAS  Google Scholar 

  13. Siedlecka, U., Arora, M., Kolettis, T., Soppa, G. K., Lee, J., Stagg, M. A., et al. (2008). Effects of clenbuterol on contractility and Ca2+ homeostasis of isolated rat ventricular myocytes. American Journal of Physiology. Heart and Circulatory Physiology, 295(5), H1917–H1926.

    Article  PubMed  CAS  Google Scholar 

  14. Mitchell, G. F., Jeron, A., & Koren, G. (1998). Measurement of heart rate and Q-T interval in the conscious mouse. American Journal of Physiology, 274(3 Pt 2), H747–H751.

    PubMed  CAS  Google Scholar 

  15. Felkin, L. E., Taegtmeyer, A. B., & Barton, P. J. (2006). Real-time quantitative polymerase chain reaction in cardiac transplant research. Methods in Molecular Biology, 333, 305–330.

    PubMed  CAS  Google Scholar 

  16. Duncker, D. J., & Bache, R. J. (2008). Regulation of coronary blood flow during exercise. Physiological Reviews, 88(3), 1009–1086.

    Article  PubMed  CAS  Google Scholar 

  17. Shiojima, I., Sato, K., Izumiya, Y., Schiekofer, S., Ito, M., Liao, R., et al. (2005). Disruption of coordinated cardiac hypertrophy and angiogenesis contributes to the transition to heart failure. Journal of Clinical Investigation, 115(8), 2108–2118.

    Article  PubMed  CAS  Google Scholar 

  18. Risuyder, J. E., Polster, S. P., Lee, S., Charles, N. J., Adhikari, N., Mariash, A., et al. (2009). Chronic treatment with clenbuterol modulates endothelial progenitor cells and circulating factors in a murine model of cardiomyopathy. Journal of Cardiovascular Translational Research, in press.

  19. Katayama, Y., Battista, M., Kao, W. M., Hidalgo, A., Peired, A. J., Thomas, S. A., et al. (2006). Signals from the sympathetic nervous system regulate hematopoietic stem cell egress from bone marrow. Cell, 124(2), 407–421.

    Article  PubMed  CAS  Google Scholar 

  20. Brancaccio, M., Hirsch, E., Notte, A., Selvetella, G., Lembo, G., & Tarone, G. (2006). Integrin signalling: the tug-of-war in heart hypertrophy. Cardiovascular Research, 70(3), 422–433.

    Article  PubMed  CAS  Google Scholar 

  21. De, A. M., Notte, A., Accornero, F., Selvetella, G., Brancaccio, M., Vecchione, C., et al. (2005). Cardiac overexpression of melusin protects from dilated cardiomyopathy due to long-standing pressure overload. Circulation Research, 96(10), 1087–1094.

    Article  Google Scholar 

  22. Shai, S. Y., Harpf, A. E., Babbitt, C. J., Jordan, M. C., Fishbein, M. C., Chen, J., et al. (2002). Cardiac myocyte-specific excision of the beta1 integrin gene results in myocardial fibrosis and cardiac failure. Circulation Research, 90(4), 458–464.

    Article  PubMed  CAS  Google Scholar 

  23. White, D. E., Coutu, P., Shi, Y. F., Tardif, J. C., Nattel, S., St, A. R., et al. (2006). Targeted ablation of ILK from the murine heart results in dilated cardiomyopathy and spontaneous heart failure. Genes & Development, 20(17), 2355–2360.

    Article  CAS  Google Scholar 

  24. Birks, E. J., Hall, J. L., Barton, P. J. R., Grindle, S., Latif, N., Hardy, J. P., et al. (2005). Gene profiling changes in cytoskeletal proteins during clinical recovery following left ventricular assist device (LVAD) support. Circulation, 112(Suppl 9), I57–I64.

    PubMed  Google Scholar 

  25. Hall, J. L., Birks, E. J., Grindle, S., Cullen, M. E., Barton, P. J., Rider, J. E., et al. (2007). Molecular signature of recovery following combination left ventricular assist device (LVAD) support and pharmacologic therapy. European Heart Journal, 28(5), 613–627.

    Article  PubMed  CAS  Google Scholar 

  26. Friddle, C. J., Koga, T., Rubin, E. M., & Bristow, J. (2000). Expression profiling reveals distinct sets of genes altered during induction and regression of cardiac hypertrophy. Proceedings of the National Academy of Sciences of the United States of America, 97(12), 6745–6750.

    Article  PubMed  CAS  Google Scholar 

  27. Dorn, I. G., Robbins, J., & Sugden, P. H. (2003). Phenotyping hypertrophy: eschew obfuscation. Circulation Research, 92(11), 1171–1175.

    Article  PubMed  CAS  Google Scholar 

  28. Ueno, S., Ohki, R., Hashimoto, T., Takizawa, T., Takeuchi, K., Yamashita, Y., et al. (2003). DNA microarray analysis of in vivo progression mechanism of heart failure. Biochemical and Biophysical Research Communications, 307(4), 771–777.

    Article  PubMed  CAS  Google Scholar 

  29. Kong, S. W., Bodyak, N., Yue, P., Liu, Z., Brown, J., Izumo, S., et al. (2005). Genetic expression profiles during physiological and pathological cardiac hypertrophy and heart failure in rats. Physiological Genomics, 21(1), 34–42.

    Article  PubMed  CAS  Google Scholar 

  30. Burniston, J. G., Tan, L. B., & Goldspink, D. F. (2005). beta2-Adrenergic receptor stimulation in vivo induces apoptosis in the rat heart and soleus muscle. Journal of Applied Physiology, 98(4), 1379–1386.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Magdi Yacoub Institute.

Author information

Authors and Affiliations

  1. Harefield Heart Science Centre, National Heart and Lung Institute, Imperial College, Hill End Road, Harefield, Middlesex, UB9 6JH, UK

    Enrique Lara-Pezzi, Cesare M. N. Terracciano, Gopal K. R. Soppa, Ryszard T. Smolenski, Leanne E. Felkin, Magdi H. Yacoub & Paul J. R. Barton

Authors
  1. Enrique Lara-Pezzi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Cesare M. N. Terracciano
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gopal K. R. Soppa
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ryszard T. Smolenski
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Leanne E. Felkin
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Magdi H. Yacoub
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Paul J. R. Barton
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Paul J. R. Barton.

Electronic supplementary material

Below is the link to the electronic supplementary material.Lara-Pezzi et al., A gene expression profile of the myocardial response to clenbuterol. Supplementary data information.

Supplementary Tables 1–4

Triplicate samples of clenbuterol treated and saline treated controls from each of the days 1 and 28 day groups were analyzed using Affymetrix DNA oligonucleotide RAE230 2.0 microchips (Affymetrix, Santa Clara, CA, USA) and data analyzed using the Genespring software suite (Agilent, Santa Clara, CA, USA) as described. Lists of genes altered in response to 1 or 28 days of clenbuterol treatment are listed; individual Excel gene lists of genes up-regulated at 1 day (Up clen 1d), down-regulated at 1 day (Down clen 1 d), or up- or down-regulated at 28 days (Up clen 28 d and Down clen 28 d respectively). (XLS 381 KB)

Supplementary Tables 5–8

Triplicate samples of clenbuterol-treated and saline-treated controls following either 1 or 28 days of treatment were analyzed using Affymetrix DNA oligonucleotide as above. Gene lists were created in which all genes showed raw expression values >100 in two out of four conditions (1 and 28 days, ±clenbuterol), flags were present or marginal in at least three samples, and gene expression showed a fold change of >1.4-fold. The resulting lists were compared to pre-existing Gene Ontology lists using Genespring Analysis (Agilent, Santa Clara, CA, USA). Resulting data are shown in Supplementary Tables 5–8 for sample sets for genes up-regulated or down-regulated at day 1 (Up clen d 1, Down clen day 1, respectively) and similarly for day 28 (Up clen 28 and Down clen 28 d) and listed under Biological process, Cellular Compartment and Molecular Function. (XLS 11.5 KB)

Supplementary Tables 9–12

Triplicate samples of clenbuterol treated and saline treated controls were analyzed using Affymetrix DNA oligonucleotide as above, and the resulting gene lists used (as defined under Supplementary Tables 5–8) analyzed using Ingenuity Pathways Analysis (Ingenuity, www.ingenuity.com). (XLS 311 KB)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lara-Pezzi, E., Terracciano, C.M.N., Soppa, G.K.R. et al. A Gene Expression Profile of the Myocardial Response to Clenbuterol. J. of Cardiovasc. Trans. Res. 2, 191–197 (2009). https://doi.org/10.1007/s12265-009-9097-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12265-009-9097-6

Keywords

Search

Navigation