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J Clin Invest. 1996 April 1; 97(7): 1577–1588.
PMCID: PMC507220

1,25(OH)2 vitamin D3, and retinoic acid antagonize endothelin-stimulated hypertrophy of neonatal rat cardiac myocytes.

Abstract

1,25(OH)2 Vitamin D3 (VD3) and retinoic acid (RA) function as ligands for nuclear receptors which regulate transcription. Though the cardiovascular system is not thought to represent a classical target for these ligands, it is clear that both cardiac myocytes and vascular smooth muscle cells respond to these agents with changes in growth characteristics and gene expression. In this study we demonstrate that each of these ligands suppresses many of the phenotypic correlates of endothelin-induced hypertrophy in a cultured neonatal rat cardiac ventriculocyte model. Each of these agents reduced endothelin-stimulated ANP secretion in a dose-dependent fashion and the two in combination proved to be more effective than either agent used alone (VD3: 49%; RA:52%; VD3 + RA:80% inhibition). RA, at concentrations known to activate the retinoid X receptor, and, to a lesser extent, VD3 effected a reduction in atrial natriuretic peptide, brain natriuretic peptide, and alpha-skeletal actin mRNA levels. Similar inhibition (VD3:30%; RA:33%; VD3 + RA:59% inhibition) was demonstrated when cells transfected with reporter constructs harboring the relevant promoter sequences were treated with VD3 and/or RA for 48 h. These effects were not accompanied by alterations in endothelin-induced c-fos, c-jun, or c-myc gene expression, suggesting either that the inhibitory locus responsible for the reduction in the mRNA levels lies distal to the activation of the immediate early gene response or that the two are not mechanistically coupled. Both VD3 and RA also reduced [3H]leucine incorporation (VD3:30%; RA:33%; VD3 + RA:45% inhibition) in endothelin-stimulated ventriculocytes and, once again, the combination of the two was more effective than either agent used in isolation. Finally, 1,25(OH)2 vitamin D3 abrogated the increase in cell size seen after endothelin treatment. These findings suggest that the liganded vitamin D and retinoid receptors are capable of modulating the hypertrophic process in vitro and that agents acting through these or similar signaling pathways may be of value in probing the molecular mechanisms underlying hypertrophy.

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Selected References

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  • Umesono K, Murakami KK, Thompson CC, Evans RM. Direct repeats as selective response elements for the thyroid hormone, retinoic acid, and vitamin D3 receptors. Cell. 1991 Jun 28;65(7):1255–1266. [PubMed]
  • Yu VC, Delsert C, Andersen B, Holloway JM, Devary OV, När AM, Kim SY, Boutin JM, Glass CK, Rosenfeld MG. RXR beta: a coregulator that enhances binding of retinoic acid, thyroid hormone, and vitamin D receptors to their cognate response elements. Cell. 1991 Dec 20;67(6):1251–1266. [PubMed]
  • Walters MR, Wicker DC, Riggle PC. 1,25-Dihydroxyvitamin D3 receptors identified in the rat heart. J Mol Cell Cardiol. 1986 Jan;18(1):67–72. [PubMed]
  • Merke J, Hofmann W, Goldschmidt D, Ritz E. Demonstration of 1,25(OH)2 vitamin D3 receptors and actions in vascular smooth muscle cells in vitro. Calcif Tissue Int. 1987 Aug;41(2):112–114. [PubMed]
  • Walters MR, Ilenchuk TT, Claycomb WC. 1,25-Dihydroxyvitamin D3 stimulates 45Ca2+ uptake by cultured adult rat ventricular cardiac muscle cells. J Biol Chem. 1987 Feb 25;262(6):2536–2541. [PubMed]
  • Weishaar RE, Simpson RU. Vitamin D3 and cardiovascular function in rats. J Clin Invest. 1987 Jun;79(6):1706–1712. [PMC free article] [PubMed]
  • Weishaar RE, Kim SN, Saunders DE, Simpson RU. Involvement of vitamin D3 with cardiovascular function. III. Effects on physical and morphological properties. Am J Physiol. 1990 Jan;258(1 Pt 1):E134–E142. [PubMed]
  • O'Connell TD, Weishaar RE, Simpson RU. Regulation of myosin isozyme expression by vitamin D3 deficiency and 1,25-dihydroxyvitamin D3 in the rat heart. Endocrinology. 1994 Feb;134(2):899–905. [PubMed]
  • Sucov HM, Dyson E, Gumeringer CL, Price J, Chien KR, Evans RM. RXR alpha mutant mice establish a genetic basis for vitamin A signaling in heart morphogenesis. Genes Dev. 1994 May 1;8(9):1007–1018. [PubMed]
  • Kastner P, Grondona JM, Mark M, Gansmuller A, LeMeur M, Decimo D, Vonesch JL, Dollé P, Chambon P. Genetic analysis of RXR alpha developmental function: convergence of RXR and RAR signaling pathways in heart and eye morphogenesis. Cell. 1994 Sep 23;78(6):987–1003. [PubMed]
  • Dyson E, Sucov HM, Kubalak SW, Schmid-Schönbein GW, DeLano FA, Evans RM, Ross J, Jr, Chien KR. Atrial-like phenotype is associated with embryonic ventricular failure in retinoid X receptor alpha -/- mice. Proc Natl Acad Sci U S A. 1995 Aug 1;92(16):7386–7390. [PubMed]
  • Baker KM, Chernin MI, Wixson SK, Aceto JF. Renin-angiotensin system involvement in pressure-overload cardiac hypertrophy in rats. Am J Physiol. 1990 Aug;259(2 Pt 2):H324–H332. [PubMed]
  • Bogoyevitch MA, Glennon PE, Andersson MB, Clerk A, Lazou A, Marshall CJ, Parker PJ, Sugden PH. Endothelin-1 and fibroblast growth factors stimulate the mitogen-activated protein kinase signaling cascade in cardiac myocytes. The potential role of the cascade in the integration of two signaling pathways leading to myocyte hypertrophy. J Biol Chem. 1994 Jan 14;269(2):1110–1119. [PubMed]
  • Simpson P, McGrath A, Savion S. Myocyte hypertrophy in neonatal rat heart cultures and its regulation by serum and by catecholamines. Circ Res. 1982 Dec;51(6):787–801. [PubMed]
  • Waspe LE, Ordahl CP, Simpson PC. The cardiac beta-myosin heavy chain isogene is induced selectively in alpha 1-adrenergic receptor-stimulated hypertrophy of cultured rat heart myocytes. J Clin Invest. 1990 Apr;85(4):1206–1214. [PMC free article] [PubMed]
  • Shubeita HE, McDonough PM, Harris AN, Knowlton KU, Glembotski CC, Brown JH, Chien KR. Endothelin induction of inositol phospholipid hydrolysis, sarcomere assembly, and cardiac gene expression in ventricular myocytes. A paracrine mechanism for myocardial cell hypertrophy. J Biol Chem. 1990 Nov 25;265(33):20555–20562. [PubMed]
  • Sadoshima J, Izumo S. Molecular characterization of angiotensin II--induced hypertrophy of cardiac myocytes and hyperplasia of cardiac fibroblasts. Critical role of the AT1 receptor subtype. Circ Res. 1993 Sep;73(3):413–423. [PubMed]
  • Duerr RL, Huang S, Miraliakbar HR, Clark R, Chien KR, Ross J., Jr Insulin-like growth factor-1 enhances ventricular hypertrophy and function during the onset of experimental cardiac failure. J Clin Invest. 1995 Feb;95(2):619–627. [PMC free article] [PubMed]
  • Sadoshima J, Jahn L, Takahashi T, Kulik TJ, Izumo S. Molecular characterization of the stretch-induced adaptation of cultured cardiac cells. An in vitro model of load-induced cardiac hypertrophy. J Biol Chem. 1992 May 25;267(15):10551–10560. [PubMed]
  • Ito H, Hirata Y, Adachi S, Tanaka M, Tsujino M, Koike A, Nogami A, Murumo F, Hiroe M. Endothelin-1 is an autocrine/paracrine factor in the mechanism of angiotensin II-induced hypertrophy in cultured rat cardiomyocytes. J Clin Invest. 1993 Jul;92(1):398–403. [PMC free article] [PubMed]
  • Arai M, Yoguchi A, Iso T, Takahashi T, Imai S, Murata K, Suzuki T. Endothelin-1 and its binding sites are upregulated in pressure overload cardiac hypertrophy. Am J Physiol. 1995 May;268(5 Pt 2):H2084–H2091. [PubMed]
  • Ito H, Hiroe M, Hirata Y, Fujisaki H, Adachi S, Akimoto H, Ohta Y, Marumo F. Endothelin ETA receptor antagonist blocks cardiac hypertrophy provoked by hemodynamic overload. Circulation. 1994 May;89(5):2198–2203. [PubMed]
  • Chien KR, Knowlton KU, Zhu H, Chien S. Regulation of cardiac gene expression during myocardial growth and hypertrophy: molecular studies of an adaptive physiologic response. FASEB J. 1991 Dec;5(15):3037–3046. [PubMed]
  • Kovacic-Milivojević B, Gardner DG. Divergent regulation of the human atrial natriuretic peptide gene by c-jun and c-fos. Mol Cell Biol. 1992 Jan;12(1):292–301. [PMC free article] [PubMed]
  • Bishopric NH, Jayasena V, Webster KA. Positive regulation of the skeletal alpha-actin gene by Fos and Jun in cardiac myocytes. J Biol Chem. 1992 Dec 15;267(35):25535–25540. [PubMed]
  • Wu J, Garami M, Cao L, Li Q, Gardner DG. 1,25(OH)2D3 suppresses expression and secretion of atrial natriuretic peptide from cardiac myocytes. Am J Physiol. 1995 Jun;268(6 Pt 1):E1108–E1113. [PubMed]
  • Li Q, Gardner DG. Negative regulation of the human atrial natriuretic peptide gene by 1,25-dihydroxyvitamin D3. J Biol Chem. 1994 Feb 18;269(7):4934–4939. [PubMed]
  • Wu J, LaPointe MC, West BL, Gardner DG. Tissue-specific determinants of human atrial natriuretic factor gene expression in cardiac tissue. J Biol Chem. 1989 Apr 15;264(11):6472–6479. [PubMed]
  • Bauer RF, Arthur LO, Fine DL. Propagation of mouse mammary tumor cell lines and production of mouse mammary tumor virus in a serum-free medium. In Vitro. 1976 Aug;12(8):558–563. [PubMed]
  • Wu JP, Deschepper CF, Gardner DG. Perinatal expression of the atrial natriuretic factor gene in rat cardiac tissue. Am J Physiol. 1988 Sep;255(3 Pt 1):E388–E396. [PubMed]
  • Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. [PubMed]
  • LaPointe MC, Sitkins JR. Phorbol ester stimulates the synthesis and secretion of brain natriuretic peptide from neonatal rat ventricular cardiocytes: a comparison with the regulation of atrial natriuretic factor. Mol Endocrinol. 1993 Oct;7(10):1284–1296. [PubMed]
  • Gustafson TA, Markham BE, Morkin E. Effects of thyroid hormone on alpha-actin and myosin heavy chain gene expression in cardiac and skeletal muscles of the rat: measurement of mRNA content using synthetic oligonucleotide probes. Circ Res. 1986 Aug;59(2):194–201. [PubMed]
  • Wu JP, Kovacic-Milivojević B, Lapointe MC, Nakamura K, Gardner DG. Cis-active determinants of cardiac-specific expression in the human atrial natriuretic peptide gene. Mol Endocrinol. 1991 Sep;5(9):1311–1322. [PubMed]
  • Seilhamer JJ, Arfsten A, Miller JA, Lundquist P, Scarborough RM, Lewicki JA, Porter JG. Human and canine gene homologs of porcine brain natriuretic peptide. Biochem Biophys Res Commun. 1989 Dec 15;165(2):650–658. [PubMed]
  • Karns LR, Kariya K, Simpson PC. M-CAT, CArG, and Sp1 elements are required for alpha 1-adrenergic induction of the skeletal alpha-actin promoter during cardiac myocyte hypertrophy. Transcriptional enhancer factor-1 and protein kinase C as conserved transducers of the fetal program in cardiac growth. J Biol Chem. 1995 Jan 6;270(1):410–417. [PubMed]
  • Baker AR, McDonnell DP, Hughes M, Crisp TM, Mangelsdorf DJ, Haussler MR, Pike JW, Shine J, O'Malley BW. Cloning and expression of full-length cDNA encoding human vitamin D receptor. Proc Natl Acad Sci U S A. 1988 May;85(10):3294–3298. [PubMed]
  • Kliewer SA, Umesono K, Heyman RA, Mangelsdorf DJ, Dyck JA, Evans RM. Retinoid X receptor-COUP-TF interactions modulate retinoic acid signaling. Proc Natl Acad Sci U S A. 1992 Feb 15;89(4):1448–1452. [PubMed]
  • Giguere V, Ong ES, Segui P, Evans RM. Identification of a receptor for the morphogen retinoic acid. Nature. 1987 Dec 17;330(6149):624–629. [PubMed]
  • Kohno M, Horio T, Yokokawa K, Yasunari K, Ikeda M, Minami M, Kurihara N, Takeda T. Brain natriuretic peptide as a marker for hypertensive left ventricular hypertrophy: changes during 1-year antihypertensive therapy with angiotensin-converting enzyme inhibitor. Am J Med. 1995 Mar;98(3):257–265. [PubMed]
  • Farach-Carson MC, Abe J, Nishii Y, Khoury R, Wright GC, Norman AW. 22-Oxacalcitriol: dissection of 1,25(OH)2D3 receptor-mediated and Ca2+ entry-stimulating pathways. Am J Physiol. 1993 Nov;265(5 Pt 2):F705–F711. [PubMed]
  • Cooper JA. Effects of cytochalasin and phalloidin on actin. J Cell Biol. 1987 Oct;105(4):1473–1478. [PMC free article] [PubMed]
  • Sharp WW, Terracio L, Borg TK, Samarel AM. Contractile activity modulates actin synthesis and turnover in cultured neonatal rat heart cells. Circ Res. 1993 Jul;73(1):172–183. [PubMed]
  • Mendelsohn C, Lohnes D, Décimo D, Lufkin T, LeMeur M, Chambon P, Mark M. Function of the retinoic acid receptors (RARs) during development (II). Multiple abnormalities at various stages of organogenesis in RAR double mutants. Development. 1994 Oct;120(10):2749–2771. [PubMed]
  • Mitsuhashi T, Morris RC, Jr, Ives HE. 1,25-dihydroxyvitamin D3 modulates growth of vascular smooth muscle cells. J Clin Invest. 1991 Jun;87(6):1889–1895. [PMC free article] [PubMed]
  • Weishaar RE, Simpson RU. The involvement of the endocrine system in regulating cardiovascular function: emphasis on vitamin D3. Endocr Rev. 1989 Aug;10(3):351–365. [PubMed]
  • Merke J, Milde P, Lewicka S, Hügel U, Klaus G, Mangelsdorf DJ, Haussler MR, Rauterberg EW, Ritz E. Identification and regulation of 1,25-dihydroxyvitamin D3 receptor activity and biosynthesis of 1,25-dihydroxyvitamin D3. Studies in cultured bovine aortic endothelial cells and human dermal capillaries. J Clin Invest. 1989 Jun;83(6):1903–1915. [PMC free article] [PubMed]
  • Zhou MD, Sucov HM, Evans RM, Chien KR. Retinoid-dependent pathways suppress myocardial cell hypertrophy. Proc Natl Acad Sci U S A. 1995 Aug 1;92(16):7391–7395. [PubMed]
  • Schräder M, Bendik I, Becker-André M, Carlberg C. Interaction between retinoic acid and vitamin D signaling pathways. J Biol Chem. 1993 Aug 25;268(24):17830–17836. [PubMed]

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