Stanozolol induces ventricular dysfunction by decreasing phospholamban phosphorylation in heart tissue of LDLr-/- mice




Anabolic androgenic steroids; Hemodynamic parameters; Cardiac remodeling; Phospholamban phosphorylation.


Stanozolol is a steroid that causes lipid deposition in LDLr-/- mice, although the mechanism by which this dyslipidemia results in cardiac dysfunction is little understood. The aim of this study was to evaluate the effect of stanozolol on cardiac contractility and the participation of myocardial phospholamban (pPLB) phosphorylation in an atherosclerosis mouse model. LDL receptor knockout mice (LDLr-/-) were fed a standard chow diet and received weekly subcutaneous injections of either saline (control, C group) or 20 mg/kg stanozolol (S group). After 8 weeks, hemodynamic parameters were assessed in the left ventricle. The heart was collected, weighted for hypertrophy evaluation, and kept in formalin buffer for morphometric analysis (H&E) and collagen quantification (Picrossirius). Protein expression of PLB and its phosphorylated form (p-PLB) in the left ventricle was determined by western blot. We observed that stanozolol treatment favored cardiac hypertrophy and collagen deposition in heart tissue. Also, stanozolol induced left ventricle dysfunction, increasing PBL expression and decreasing the p-PLB/PLB ratio. Altogether, our data showed that stanozolol causes cardiac remodeling and ventricular dysfunction by decreasing PLB phosphorylation in the left ventricle of LDLr-/- mice.


Almeida, S. A. de, Claudio, E. R. G., Mengal, V., Brasil, G. A., Merlo, E., Podratz, P. L., … Abreu, G. R. de. (2018). Estrogen Therapy Worsens Cardiac Function and Remodeling and Reverses the Effects of Exercise Training After Myocardial Infarction in Ovariectomized Female Rats. Frontiers in Physiology: Exercise Physiology, 9, 1–11.

Almeida, S. A. De, Claudio, E. R. G., Mengal, V. F., Oliveira, S. G. De, Merlo, E., Podratz, P. L., … de Abreu, G. R. (2014). Exercise Training Reduces Cardiac Dysfunction and Remodeling in Ovariectomized Rats Submitted to Myocardial Infarction. PloS One, 9(12), e115970.

Ammar, E. M., Said, S. A., & Hassan, M. S. (2004). Enhanced vasoconstriction and reduced vasorelaxation induced by testosterone and nandrolone in hypercholesterolemic rabbits, 50, 253–259.

Andrade, Tadeu Uggere de, Haguihara, S. C. G. C., Falsoni, R. M. P., da Silva, C. L., Dubois Filho, D. G., de Souza Andrade Moraes, F., … de Lima, E. M. (2019). Stanozolol promotes lipid deposition in the aorta through an imbalance in inflammatory cytokines and oxidative status in LDLr knockout mice fed a normal diet. Basic and Clinical Pharmacology and Toxicology, 124(4), 360–369.

Andrade, Tadeu Uggere De, Loiola, L. Z., Merces, S., Alcure, N., Raquel, A., Medeiros, S., … Bissoli, N. S. (2011). Role of the renin – angiotensin system in the nandrolone-decanoate-induced attenuation of the Bezold – Jarisch reflex. Canadian Journal of Physiology and Pharmacology, 897, 891–897.

Andrade, Tadeu U, Santos, C. S., Busato, C. W., Medeiros, R. S., Abreu, R., & Moyse, M. R. (2008). Higher Physiological Doses of Nandrolone Decanoate Do Not Influence the Bezold-Jarish Reflex Control of Bradycardia, 39, 27–32.

Beutel, A., Bergamaschi, T., & Campos, R. R. (2005). Effects of chronic anabolic steroid treatment on tonic and reflex cardiovascular control in male rats. Steroid Biochemistry & Molecular Biology, 93, 43–48.

Bozi, L., Maldonado, I., Baldo, M., Silva, M., Moreira, J., Novaes, R., … Natali, A. (2013). Exercise training prior to myocardial infarction attenuates cardiac deterioration and cardiomyocyte dysfunction in rats. Clinics, 68(4), 549–556.

Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72(1–2), 248–254.

Brasil, G. A., Lima, E. M. De, Nascimento, A. M. do, Caliman, I. F., Medeiros, A. R. S. de, Silva, M. S. B., … Bissoli, N. S. (2015). Nandrolone decanoate induces cardiac and renal remodeling in female rats, without modification in physiological parameters: The role of ANP system. Life Sciences, 137, 65–73.

Chagwon, K., Ahyoung, L., & Roger, J. H. (2013). Altered sarcoplasmic reticulum calcim cycling-targets for heart failure therapy. Nature Reviews Cardiology, 9(12), 717–733.

Chaves, E. A., Pereira-Junior, P. P., Fortunato, R. S., Masuda, M. O., Carvalho, A. C. C. de, Carvalho, D. P. de, … Nascimento, J. H. M. (2006). Nandrolone decanoate impairs exercise-induced cardioprotection : Role of antioxidant enzymes. The Journal of Steroid Biochemistry & Molecular Biology, 99, 223–230.

Darke, S., Torok, M., & Duflou, J. (2014). Sudden or Unnatural Deaths Involving Anabolic-androgenic Steroids. Journal of Forensic Sciences, 59(4).

Davies, C. H., Harding, S. E., & Poole-Wilson, P. A. (1996). Cellular mechanisms of contractile dysfunction in human heart failure. European Heart Journal, 17(2), 189–198.

Davis, K. L., Mehlhorn, U., Schertel, E. R., Geissler, H. J., Trevas, D., Laine, G. A., & Allen, S. J. (1999). Variation in Tau, the time constant for isovolumic relaxation, along the left ventricular base-to-apex axis. Basic Research in Cardiology, 94(1), 41–48.

De Andrade, Tadeu Uggere, Abreu, G. R., Moysés, M. R., De Melo Cabral, A., & Bissoli, N. S. (2008). Role of cardiac hypertrophy in reducing the sensitivity of cardiopulmonary reflex control of renal sympathetic nerve actvity in spontaneously hypertensive rats. Clinical and Experimental Pharmacology and Physiology, 35(9), 1104–1108.

El-Mas, M. M. (2002). Cyclosporine Adversely Affects Baroreflexes via Inhibition of Testosterone Modulation of Cardiac Vagal Control. Journal of Pharmacology and Experimental Therapeutics, 301(1), 346–354.

El-Mas, M. M., Afify, E. A., Mohy El-Din, M. M., Omar, A. G., & Sharabi, F. M. (2001). Testosterone facilitates the baroreceptor control of reflex bradycardia: Role of cardiac sympathetic and parasympathetic components. Journal of Cardiovascular Pharmacology, 38(5), 754–763.

Ewart, M. A., Kennedy, S., MacMillan, D., Raja, A. L. N., Watt, I. M., & Currie, S. (2014). Altered vascular smooth muscle function in the ApoE knockout mouse during the progression of atherosclerosis. Atherosclerosis, 234(1), 154–161.

Fernandes, T., Soci, U. P. R., & Oliveira, E. M. (2011). Eccentric and concentric cardiac hypertrophy induced by exercise training: microRNAs and molecular determinants. Brazilian Journal of Medical and Biological Research, 44(September), 836–847.

Fogelberg, M., Björkhem, I., Diczfalusy, U., & Henriksson, P. (1990). Stanozolol and experimental atherosclerosis: Atherosclerotic development and blood lipids during anabolic steroid therapy of New zealand white rabbits. Scandinavian Journal of Clinical and Laboratory Investigation, 50(6), 693–696.

Fontana, K., Oliveira, H. C. F., Leonardo, M. B., Mandarim-de-Lacerda, C. A., & Cruz-Hofling, M. A. da. (2008). Adverse effect of the anabolic-androgenic steroid mesterolone on cardiac remodelling and lipoprotein profile is attenuated by aerobicz exercise training. International Journal of Experimental Pathology, 89(5), 358–366.

Franquni, J. V. M., Nascimento, A. M. do, Lima, E. M. de, Brasil, G. A., Heringer, O. A., Cassaro, K. O. dos S., … Andrade, T. U. de. (2013). Nandrolone decanoate determines cardiac remodelling and injury by an imbalance in cardiac inflammatory cytokines and ACE activity , blunting of the Bezold – Jarisch reflex , resulting in the development of hypertension. Steroids, 78, 379–385.

Garcia, J. A. D., Dos Santos, L., Moura, A. L., Ricardo, K. F. S., Wanschel, A. C. B. A., Shishido, S. M., … Krieger, M. H. (2008). S-nitroso-N-acetylcysteine (SNAC) prevents myocardial alterations in hypercholesterolemic LDL receptor knockout mice by antiinflammatory action. Journal of Cardiovascular Pharmacology, 51(1), 78–85.

Gardner, J. D., Brower, G. L., & Janicki, J. S. (2002). Gender differences in cardiac remodeling secondary to chronic volume overload. Journal of Cardiac Failure, 8(2), 101–107.

Gleason, W. L., & Braunwald, E. (1962). Studies on the first derivative of the ventricular pressure pulse in man. The Journal of Clinical Investigation, 41(1), 80–91.

Henning, R. J., Cheng, J., & Levy, M. N. (1989). Vagal stimulation decreases rate of left ventricular relaxation. American Journal of Physiology-Heart and Circulatory Physiology, 256(2), H428–H433.

Henning, R. J., Khalil, I. R., & Levy, M. N. (1990). Vagal stimulation attenuates sympathetic enhancement of left ventricular function. American Journal of Physiology-Heart and Circulatory Physiology, 258(5), H1470–H1475.

Henning, Robert J, & Levy, M. N. (1991). Effects of autonomic nerve stimulation, asynchrony, and load on dP/dt max and on dP/dt min. American Physiological Society.

Herrington, W., Lacey, B., Sherliker, P., Armitage, J., & Lewington, S. (2016). Epidemiology of Atherosclerosis and the Potential to Reduce the Global Burden of Atherothrombotic Disease. Circulation Research, 118(4), 535–546.

Huang, B., Wang, S., Qin, D., Boutjdir, M., & El-Sherif, N. (1999). Diminished Basal Phosphorylation Level of Phospholamban in the Postinfarction Remodeled Rat Ventricle. Circulation Research, 85(9), 848–855.

James, P., Tada, M., Chiesit, M., & Carafoli, E. (1989). Nature and site of phospholamban regulation of the Ca2+ pump of sarcoplasmic reticulum. Letters to Nature, 342(November), 0–2.

Junqueira, L. C. U., Bignolas, G., & Brentani, R. R. (1979). Picrosirius Staining Plus Polarization Microscopy, a Specific Method for Collagen Detection in Tissue-Sections. Histochemical Journal, 11(4), 447–455.

Kho, C., Lee, A., & Hajjar, R. J. (2012). Altered sarcoplasmic reticulum calcium cycling - Targets for heart failure therapy. Nature Reviews Cardiology, 9(12), 717–733.

Kicman, A. T. (2008). Pharmacology of anabolic steroids. British Journal of Pharmacology, 17(February), 502–521.

Kuhn, C. M. (2002). Anabolic Steroids. Recent Progress in Hormone Research, 411–434.

Larsen, K.-O., Sjaastad, I., Svindland, A., Krobert, K. A., Skjønsberg, O. H., & Christensen, G. (2006). Alveolar hypoxia induces left ventricular diastolic dysfunction and reduces phosphorylation of phospholamban in mice. American Journal of Physiology-Heart and Circulatory Physiology, 291(2), H507–H516.

Lee, Y. T., Lin, H. Y., Chan, Y. W. F., Li, K. H. C., To, O. T. L., Yan, B. P., … Tse, G. (2017). Mouse models of atherosclerosis: a historical perspective and recent advances. Lipids in Health and Disease, 16(1), 12.

Leite-Moreira, A. F., & Gillebert, T. C. (1994). Nonuniform course of left ventricular pressure fall and its regulation by load and contractile state. Circulation, 90(5), 2481–2491.

Lima, E. M. E. M., Nascimento, A. M. A. M., Brasil, G. A. G. A., Kalil, I. C. I. C., Lenz, D., Endringer, D. C. D. C., … Bissoli, N. S. N. S. (2015). Cardiopulmonary reflex, cardiac cytokines, and nandrolone decanoate: response to resistance training in rats. Canadian Journal of Physiology and Pharmacology, 7(May), 1–7.

Lindemann, J. P., & Watanabe, A. M. (1985). Muscarinic cholinergic inhibition of ??-adrenergic stimulation of phospholamban phosphorylation and Ca2+ transport in guinea pig ventricles. Journal of Biological Chemistry, 260(24), 13122–13129.

Lippi, G., Franchini, M., & Banfi, G. (2011). Biochemistry and Physiology of Anabolic Androgenic Steroids Doping. Mini-Reviews in Medicinal Chemistry, 11, 362–373.

Litwin, S. E., Raya, T. E., Anderson, P. G., Litwin, C. M., Bressler, R., & Goldman, S. (1991). Induction of myocardial hypertrophy after coronary ligation in rats decreases ventricular dilatation and improves systolic function. Circulation, 84(4), 1819–1827.

Liu, P. Y., Death, A. K., & Handelsman, D. J. (2003). Androgens and cardiovascular disease. Endocrine Reviews, 24(3), 313–340.

Marques-neto, S. R., Ferraz, E. B., Rodrigues, D. C., Njaine, B., Rondinelli, E., Carvalho, A. C. C. de, & Nascimento, J. H. M. (2014). AT1 and Aldosterone Receptors Blockade Prevents the Chronic Effect of Nandrolone on the Exercise-Induced Cardioprotection in Perfused rat Heart Subjected to Ischemia and Reperfusion. Cardiovascular Drugs and Therapy, 28, 125–135.

Mattiazzi, A., Garay, A., & Cingolani, H. E. (1986). Critical evaluation of isometric indexes of relaxation in rat and cat papillary muscles and toad ventricular strips. Journal of Molecular and Cellular Cardiology, 18(7), 749–758.

Melo Junior, A. F., Dalpiaz, P. L. M., Sousa, G. J., Oliveira, P. W. C., Birocale, A. M., Andrade, T. U., … Bissoli, N. S. (2018). Nandrolone alter left ventricular contractility and promotes remodelling involving calcium-handling proteins and renin-angiotensin system in male SHR. Life Sciences, 208(May), 239–245.

Mishra, S., Bedja, D., Amuzie, C., Avolio, A., & Chatterjee, S. (2015). Prevention of Cardiac Hypertrophy by the Use of a Glycosphingolipid Synthesis Inhibitor in ApoE−/− Mice. Biochemical and Biophysical Research Communications, 91(2), 165–171.

Mozaffarian, D., Benjamin, E. J., Go, A. S., Arnett, D. K., Blaha, M. J., Cushman, M., … Turner, M. B. (2016). AHA Statistical Update Heart Disease and Stroke Statistics — 2016 Update A Report From the American Heart Association. Circulation, 133, e38–e360.

Nascimento, A. M. do, Lima, E. M. de, Brasil, G. A., Caliman, I. F., Silva, J. F. da, Lemos, V. S., … Bissoli, N. S. (2016). Serca2a and Na + /Ca 2 + exchanger are involved in left ventricular function following cardiac remodelling of female rats treated with anabolic androgenic steroid. Toxicology and Applied Pharmacology, 301, 22–30.

Norton, G. R., Trifunovic, B., & Woodiwiss, A. J. (2000). Attenueted β-adrenoceptor-mediated cardiac contractile responses following androgenic steroid administration to sedentary rats. European Journal of Applied Physiology, 81(4), 310–316.

Paolo, M. Di, Agozzino, M., Toni, C., Luciani, A. B., Molendini, L., Scaglione, M., … Arbustini, E. (2007). Sudden anabolic steroid abuse-related death in athletes. International Journal of Cardiology, 114, 114–117.

Pfeffer, M. A., Pfeffer, J. M., Fishbein, M. C., Fletcher, P. J., Spadaro, J., Kloner, R. A., & Braunwald, E. (1979). Myocardial infarct size and ventricular function in rats. Circ Res, 44(4), 503–512.

Seto, S.-W., Krishna, S. M., Moran, C. S., Liu, D., & Golledge, J. (2014). Aliskiren limits abdominal aortic aneurysm, ventricular hypertrophy and atherosclerosis in an apolipoprotein-E-deficient mouse model. Clinical Science, 127(2), 123–134.

Silva, D., Miranda, A., Silva, D., D`Angelo, L., Rosa, B., Soares, E., … Garcia, J. (2015). Propolis and swimming in the prevention of atherogenesis and left ventricular hypertrophy in hypercholesterolemic mice. Brazilian Journal of Biology, 75(2), 414–422.

Simmerman, H. K., & Jones, L. R. (2017). Phospholamban: Protein Structure, Mechanism of Action, and Role in Cardiac Function. Physiological Reviews, 78(4), 921–947.

Smith, P. J. W., Ornatsky, O., Stewart, D. J., Picard, P., Dawood, F., Wen, W.-H., … Monge, J. C. (2000). Effects of Estrogen Replacement on Infarct Size, Cardiac Remodeling, and the Endothelin System After Myocardial Infarction in Ovariectomized Rats. Circulation, 102(24), 2983–2989.

Sutton, M. G. S. J., & Sharpe, N. (2000). Left Ventricular Remodeling After Myocardial Infarction: Pathophysiology and Therapy. Circulation, 101, 2981–2988.

Tanno, A. P., Neves, V. J. das, Rosa, K. T., Cunha, T. S., Giordano, F. C. L., Calil, C. M., … Marcondes, F. K. (2011). Nandrolone and resistance training induce heart remodeling: Role of fetal genes and implications for cardiac pathophysiology. Life Sciences, 89(17–18), 631–637.

Veseli, B. E., Perrotta, P., Meyer, G. R. A. De, Roth, L., Donckt, C. Van der, Martinet, W., & Meyer, G. R. Y. De. (2017). Animal models of atherosclerosis. European Journal of Pharmacology, (April), 1–11.

Viana Gonçalves, I. C., Cerdeira, C. D., Poletti Camara, E., Dias Garcia, J. A., Ribeiro Pereira Lima Brigagão, M., Bessa Veloso Silva, R., & Bitencourt dos Santos, G. (2017). Tempol improves lipid profile and prevents left ventricular hypertrophy in LDL receptor gene knockout (LDLr-/-) mice on a high-fat diet. Revista Portuguesa de Cardiologia, 36(9), 629–638.




How to Cite

ANDRADE, T. U. de .; DUBOIS FILLHO, D.; SILVA, C. L. da .; SILVA, M. de A. .; ALMEIDA, S. A. de .; NASCIMENTO, A. M. do .; BISSOLI, N. S. .; BRASIL, G. A.; LIMA, E. M. de . Stanozolol induces ventricular dysfunction by decreasing phospholamban phosphorylation in heart tissue of LDLr-/- mice . Research, Society and Development, [S. l.], v. 11, n. 5, p. e12911527876, 2022. DOI: 10.33448/rsd-v11i5.27876. Disponível em: Acesso em: 3 mar. 2024.



Health Sciences