Abstract
The aim of this work is to study cytoskeletal impairment during the development of ouabain-induced ventricular hypertrophy. Male Sprague–Dawley rats were treated with either ouabain or saline. Systolic blood pressure (SBP) was recorded weekly. At the end of the 3rd and 6th week, the rats were killed and cardiac mass index were measured. Hematoxylin–eosin and Sirius red staining were carried out and cardiac ultrastructure were studied using transmission electron microscopy. The mRNA level of Profilin-1, Desmin, PCNA, TGF-β1 and ET-1 in the left ventricle were measured using real-time quantitative PCR while their protein levels were examined by Western blot or immunohistochemistry. After 3 weeks, there was no significant difference in the mean SBP, cardiac mass index, mRNA and protein expression of PCNA, TGF-β1 and ET-1 between the two groups. However, ouabain-treated rats showed disorganized cardiac cytoskeleton with abnormal expression of Profilin-1 and Desmin. After 6 weeks, the cardiac mass index remained the same in the two groups while PCNA, TGF-β1, and ET-1 have been upregulated in ouabain-treated rats. The cardiac cytoskeletal impairment was more severe in ouabain-treated rats with further changes of Profilin-1 and Desmin. Cytoskeletal abnormality is an ultra-early change during ouabain-induced ventricular hypertrophy, before the release of hypertrophic factors. Therapy for prevention of ouabain-induced hypertrophy should start at the early stage by preventing the cytoskeleton from disorganization.










Similar content being viewed by others
References
Xie Z, Cai T (2003) Na+-K+-ATPase-mediated signal transduction: from protein interaction to cellular function. Mol Interv 3:157–168
Schoner W, Scheiner-Bobis G (2007) Endogenous and exogenous cardiac glycosides and their mechanisms of action. Am J Cardiovasc Drugs 7:173–189
Manunta P, Ferrandi M, Bianchi G, Hamlyn JM (2009) Endogenous ouabain in cardiovascular function and disease. J Hypertens 27:9–18
Ferrandi M, Molinari I, Barassi P, Minotti E, Bianchi G, Ferrari P (2004) Organ hypertrophic signaling within caveolae membrane subdomains triggered by ouabain and antagonized by PST 2238. J Biol Chem 279:33306–33314
Kometiani P, Li J, Gnudi L, Kahn BB, Askari A, Xie Z (1998) Multiple signal transduction pathways link Na+/K+-ATPase to growth-related genes in cardiac myocytes. The roles of Ras and mitogen-activated protein kinases. J Biol Chem 273:15249–15256
Huang L, Li H, Xie Z (1997) Ouabain-induced hypertrophy in cultured cardiac myocytes is accompanied by changes in expression of several late response genes. J Mol Cell Cardiol 29:429–437
Liu J, Tian J, Haas M, Shapiro JI, Askari A, Xie Z (2000) Ouabain interaction with cardiac Na+/K+-ATPase initiates signal cascades independent of changes in intracellular Na+ and Ca2+ concentrations. J Biol Chem 275:27838–27844
Collins JF, Pawloski-Dahm C, Davis MG, Ball N, Dorn GW 2nd, Walsh RA (1996) The role of the cytoskeleton in left ventricular pressure overload hypertrophy and failure. J Mol Cell Cardiol 28:1435–1443
Qiu J, Gao HQ, Zhou RH, Liang Y, Zhang XH, Wang XP, You BA, Cheng M (2007) Proteomics analysis of the proliferative effect of low-dose ouabain on human endothelial cells. Biol Pharm Bull 30:247–253
Moustafa-Bayoumi M, Alhaj MA, El-Sayed O, Wisel S, Chotani MA, Abouelnaga ZA, Hassona MD, Rigatto K, Morris M, Nuovo G, Zweier JL, Goldschmidt-Clermont P, Hassanain H (2007) Vascular hypertrophy and hypertension caused by transgenic overexpression of profilin 1. J Biol Chem 282:37632–37639
Kim HR, Graceffa P, Ferron F, Gallant C, Boczkowska M, Dominguez R, Morgan KG (2010) Actin polymerization in differentiated vascular smooth muscle cells requires vasodilator-stimulated phosphoprotein. Am J Physiol Cell Physiol 298:C559–C571
Costa ML, Escaleira R, Cataldo A, Oliveira F, Mermelstein CS (2004) Desmin: molecular interactions and putative functions of the muscle intermediate filament protein. Braz J Med Biol Res 37:1819–1830
Tolstonog GV, Sabasch M, Traub P (2002) Cytoplasmic intermediate filaments are stably associated with nuclear matrices and potentially modulate their DNA-binding function. DNA Cell Biol 21:213–239
Monreal G, Nicholson LM, Han B, Joshi MS, Phillips AB, Wold LE, Bauer JA, Gerhardt MA (2008) Cytoskeletal remodeling of desmin is a more accurate measure of cardiac dysfunction than fibrosis or myocyte hypertrophy. Life Sci 83:786–794
Yuan CM, Manunta P, Hamlyn JM, Chen S, Bohen E, Yeun J, Haddy FJ, Pamnani MB (1993) Long-term ouabain administration produces hypertension in rats. Hypertension 22:178–187
Tian G, Dang C, Lu Z (2001) The change and significance of the Na+-K+-ATPase alpha-subunit in ouabain-hypertensive rats. Hypertens Res 24:729–734
Hamlyn JM, Manunta P (1992) Ouabain, digitalis-like factors and hypertension. J Hypertens Suppl 10:S99–S111
Lewis LK, Yandle TG, Lewis JG, Richards AM, Pidgeon GB, Kaaja RJ, Nicholls MG (1994) Ouabain is not detectable in human plasma. Hypertension 24:549–555
Gruson D, Ginion A, Decroly N, Lause P, Vanoverschelde JL, Ketelslegers JM, Bertrand L, Thissen JP (2011) Urocortin-induced cardiomyocytes hypertrophy is associated with regulation of the GSK-3beta pathway. Heart Vessels. doi:10.1007/s00380-011-0141-5
Kamal FA, Watanabe K, Ma M, Abe Y, Elbarbary R, Kodama M, Aizawa Y (2011) A novel phenylpyridazinone, T-3999, reduces the progression of autoimmune myocarditis to dilated cardiomyopathy. Heart Vessels 26:81–90
Manunta P, Stella P, Rivera R, Ciurlino D, Cusi D, Ferrandi M, Hamlyn JM, Bianchi G (1999) Left ventricular mass, stroke volume, and ouabain-like factor in essential hypertension. Hypertension 34:450–456
Jiang X, Ren YP, Lv ZR (2007) Ouabain induces cardiac remodeling in rats independent of blood pressure. Acta Pharmacol Sin 28:344–352
Manunta P, Rogowski AC, Hamilton BP, Hamlyn JM (1994) Ouabain-induced hypertension in the rat: relationships among plasma and tissue ouabain and blood pressure. J Hypertens 12:549–560
Capetanaki Y (2000) Desmin cytoskeleton in healthy and failing heart. Heart Fail Rev 5:203–220
Lesnefsky EJ, Moghaddas S, Tandler B, Kerner J, Hoppel CL (2001) Mitochondrial dysfunction in cardiac disease: ischemia–reperfusion, aging, and heart failure. J Mol Cell Cardiol 33:1065–1089
Liu T, Brown DA, O’Rourke B (2010) Role of mitochondrial dysfunction in cardiac glycoside toxicity. J Mol Cell Cardiol 49:728–736
Yarmola EG, Bubb MR (2006) Profilin: emerging concepts and lingering misconceptions. Trends Biochem Sci 31:197–205
Machesky LM, Poland TD (1993) Profilin as a potential mediator of membrane-cytoskeleton communication. Trends Cell Biol 3:381–385
Romeo GR, Kazlauskas A (2008) Oxysterol and diabetes activate STAT3 and control endothelial expression of profilin-1 via OSBP1. J Biol Chem 283:9595–9605
Si J, Collins SJ (2008) Activated Ca2+/calmodulin-dependent protein kinase IIgamma is a critical regulator of myeloid leukemia cell proliferation. Cancer Res 68:3733–3742
Onishi A, Chen Q, Humtsoe JO, Kramer RH (2008) STAT3 signaling is induced by intercellular adhesion in squamous cell carcinoma cells. Exp Cell Res 314:377–386
Takekoshi K, Ishii K, Kawakami Y, Isobe K, Nanmoku T, Nakai T (2001) Ca(2+) mobilization, tyrosine hydroxylase activity, and signaling mechanisms in cultured porcine adrenal medullary chromaffin cells: effects of leptin. Endocrinology 142:290–298
Heling A, Zimmermann R, Kostin S, Maeno Y, Hein S, Devaux B, Bauer E, Klovekorn WP, Schlepper M, Schaper W, Schaper J (2000) Increased expression of cytoskeletal, linkage, and extracellular proteins in failing human myocardium. Circ Res 86:846–853
Milner DJ, Weitzer G, Tran D, Bradley A, Capetanaki Y (1996) Disruption of muscle architecture and myocardial degeneration in mice lacking desmin. J Cell Biol 134:1255–1270
Chen F, Chang R, Trivedi M, Capetanaki Y, Cryns VL (2003) Caspase proteolysis of desmin produces a dominant-negative inhibitor of intermediate filaments and promotes apoptosis. J Biol Chem 278:6848–6853
Milner DJ, Taffet GE, Wang X, Pham T, Tamura T, Hartley C, Gerdes AM, Capetanaki Y (1999) The absence of desmin leads to cardiomyocyte hypertrophy and cardiac dilation with compromised systolic function. J Mol Cell Cardiol 31:2063–2076
El-Ani D, Stav H, Guetta V, Arad M, Shainberg A (2011) Rapamycin (sirolimus) protects against hypoxic damage in primary heart cultures via Na+/Ca2+ exchanger activation. Life Sci 89:7–14
Maruyama R, Takemura G, Tohse N, Ohkusa T, Ikeda Y, Tsuchiya K, Minatoguchi S, Matsuzaki M, Fujiwara T, Fujiwara H (2006) Synchronous progression of calcium transient-dependent beating and sarcomere destruction in apoptotic adult cardiomyocytes. Am J Physiol Heart Circ Physiol 290:H1493–H1502
Weisleder N, Soumaka E, Abbasi S, Taegtmeyer H, Capetanaki Y (2004) Cardiomyocyte-specific desmin rescue of desmin null cardiomyopathy excludes vascular involvement. J Mol Cell Cardiol 36:121–128
Milner DJ, Mavroidis M, Weisleder N, Capetanaki Y (2000) Desmin cytoskeleton linked to muscle mitochondrial distribution and respiratory function. J Cell Biol 150:1283–1298
Capetanaki Y (2002) Desmin cytoskeleton: a potential regulator of muscle mitochondrial behavior and function. Trends Cardiovasc Med 12:339–348
(1997) Uniform requirements for manuscripts submitted to biomedical journals. International Committee of Medical Journal Editors. N Engl J Med 336:309–315
Acknowledgments
This study was supported by grants from the National Nature Science Foundation of China (30700884), Shandong Science and Technology Research Plan (2010GGC10294) and Shandong Outstanding Young and Middle-aged Scientists Research Award Fund (BS2009SW015) to Dr. Jie Qiu. The authors of this manuscript have certified that they comply with the Uniform Requirements for Manuscripts Submitted to Biomedical Journals [42].
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zhao, Sh., Gao, Hq., Ji, X. et al. Effect of ouabain on myocardial ultrastructure and cytoskeleton during the development of ventricular hypertrophy. Heart Vessels 28, 101–113 (2013). https://doi.org/10.1007/s00380-011-0219-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00380-011-0219-0