Abstract
Earlier studies have demonstrated angiotensin II (AngII) and aldosterone (ALDO) each augment cultured adult rat cardiac fibroblast (CFb) collagen synthesis. Whether this involves type I collagen, the major structural protein of the myocardium, and represents a transcriptional event, is uncertain. Accordingly, the influence of AngII and ALDO on transcription and synthesis of fibrillar collagen and on collagenolytic activity was examined in cultured CFb maintained in serum-deprived media. Using concentrations for AngII (10−7 M) or ALDO (10−9 M), shown to influence collagen turnover in these cells, we found: a) total collagen synthesis was significantly (p < 0.05) increased (5.4 ± 0.41 and 4.8 ± 0.37 vs. control 3.1 ± 0.55); b) type I collagen production (6590 ± 710 and 6150 ± 410 vs. control 4700 ± 490 ng/mL) in the medium were significantly (p < 0.01) increased; c) type I collagen mRNA expression was also significantly (p < 0.01) increased by AngII (2.0 fold) and ALDO (1.8 fold) compared with control; d) AngII, but not ALDO, significantly (p < 0.05) decreased collagenolytic activity (0.5 fold) compared with control. Thus, AngII and ALDO each increase CFb type I collagen synthesis at the level of transcription and protein synthesis and AngII, but not ALDO, alters collagenolytic activity. Such hormonally mediated alterations in CFb collagen turnover may contribute to the adverse accumulation of fibrillar collagen found in the myocardium in various disease states, where circulating AngII and/or ALDO are increased.
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Weber KT, Janicki JS, Pick R, Capasso J, Anversa P: Myocardial fibrosis and pathologic hypertrophy in the rat with renovascular hypertension. Am J Cardiol 65: 1G-7G, 1990
Nath KA: Tubulointerstitial changes as a major determinant in the progression of renal damage. Am J Kidney Dis 20: 1–17, 1992
Nakano M, Lieber CS: Ultrastructure of initial stages of perivascular fibrosis in alcohol-fed baboons. Am J Pathol 106: 145–155, 1982
Phan SH, Thrall RS, Williams C: Bleomycin-induced pulmonary fibrosis. Effects of steroid on lung collagen metabolism. Am Rev Respir Dis 124: 428–434, 1981
Chua CC, Chua BHL, Zhao ZY, Krebs C, Diglio C, Perrin E: Effect of growth factors on collagen metabolism in cultured human heart fibroblasts. Connect Tissue Res 26: 271–281, 1991
Bashey RI, Donnelly M, Insigna F, Jimenez SA: Growth properties and biochemical characterization of collagens synthesized by adult rat heart fibroblasts in culture. J Mol Cell Cardiol 24: 691–700, 1992
Eghbali M, Czaja MJ, Zeyel M, Weiner FR, Zern MA, Seifter S, Blumenfeld OO: Collagen chain mRNAs in isolated heart cells from young and adult rats. J Mol Cell Cardiol 20: 267–276, 1988
Brilla CG, Zhou G, Matsubara L, Weber KT: Collagen metabolism in cultured adult rat cardiac fibroblasts: response to angiotensin II and aldosterone. J Mol Cell Cardiol 26: 809–820, 1994
Wolf G, Haberstroh U, Neilson EG: Angiotensin II stimulates the proliferation and biosynthesis of type I collagen in cultured murine mesangial cells. Am J Pathol 140: 95–107, 1992
Muller GA, Markovic-Lipkovski J, Rodemann HP: The progression of renal diseases: on the pathogenesis of renal interstitial fibrosis. Klin Wochenschr 69: 576–586, 1991
Friedman SL: The cellular basis of hepatic fibrosis. Mechanisms and treatment strategies. N Engl J Med 328: 1828–1835, 1993
Price MG: Molecular analysis of intermediate filament cytoskeleton: a putative load-bearing structure. Am J Physiol 15: H566-H572, 1984
Dostal DE, Rothblum KN, Chernin MI, Cooper GR, Baker KM: Intracardiac detection of angiotensinogen and renin: a localized renin-angiotensin system in neonatal rat heart. Am J Physiol 263: C838-C850, 1992
Dostal DE, Rothblum KN, Conrad KM, Cooper GR, Baker KM: Detection of angiotensin I and II in cultured rat cardiac myocytes and fibroblasts. Am J Physiol 263: C851-C863, 1992
Schorb W, Booz GW, Dostal DE, Conrad KM, Chang KC, Baker KM: Angiotensin II is mitogenic in neonatal rat cardiac fibroblasts. Circ Res 72: 1245–1254, 1993
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 73: 413–423, 1993
Adzick NS, Longaker MT: Scarless fetal healing. Therapeutic implications. Ann Surg 215: 3–7, 1992
Eghbali M, Blumenfeld OO, Seifter S, Buttrick PM, Leinwand LA, Robinson TF, Zern MA, Giambrone MA: Localization of types I, III, and IV collagen mRNAs in rat heart cells by in situ hybridization. J Mol Cell Cardiol 21: 103–113, 1989
Brenner DA, Westwick J, Breindl M: Type I collagen gene regulation and the molecular pathogenesis of cirrhosis. Am J Physiol 264: G589-G595, 1993
Medugorac I: Collagen content in different areas of normal and hypertrophied rat myocardium. Cardiovasc Res 14: 551–554, 1980
Weber KT, Janicki JS, Shroff SG, Pick R, Chen RM, Bashey RI: Collagen remodeling of the pressure-overloaded, hypertrophied nonhuman primate myocardium. Circ Res 62: 757–765, 1988
Bishop J, Greenbaum J, Gibson D, Yacoub M, Laurent GJ: Enhanced deposition of predominantly type I collagen in myocardial disease. J Mol Cell Cardiol 22: 1157–1165, 1990
Mukherjee D, Sen S: Collagen phenotypes during development and regression of myocardial hypertrophy in spontaneously hypertensive rats. Circ Res 67: 1474–1480, 1990
Bishop JE, Mitchell JJ, Absher PM, Baldor L, Geller HA, Woodcock-Mitchell J, Hamblin MJ, Vacek P, Low RB: Cyclic mechanical deformation stimulates human lung fibroblast proliferation and autocrine growth factor activity. Am J Respir Cell Mol Biol 9: 126–133, 1993
Harrison NK, Argent AC, McAnulty RJ, Black CM, Corrin B, Laurent GJ: Collagen synthesis and degradation by systemic lung fibroblasts. Responses to transforming growth factor-beta. Chest 99, Suppl 3: 715–725, 1991
Laurent GJ: Dynamic state of collagen: pathways of collagen degradation in vivo and their possible role in regulation of collagen mass. Am J Physiol 252: C1-C9, 1987
Weber KT, Brilla CG: Pathological hypertrophy and cardiac interstitium: fibrosis and renin-angiotensin-aldosterone system. Circulation 83: 1849–1865, 1991
Villarreal FJ, Kim NN, Ungab GD, Printz MP, Dillman WH: Identification of functional angiotensin II receptors on rat cardiac fibroblasts. Circulation 88: 2849–2861, 1993
Diglio CA, Grammas P, Giacomelli F, Wiener J: Rat heart-derived endothelial and smooth muscle cell cultures: isolation, cloning and characterization. Tissue Cell 20: 477–492, 1988
Peterkofsky B, Diegelmann R: Use of a mixture of proteinase-free collagenases for the specific assay of radioactive collagen in the presence of other proteins. Biochemistry 10: 988–994, 1971
Bersohn MM, Scheuer J: Effects of physical training on end-diastolic volume and myocardial performance of isolated rat hearts. Circ Res 40: 510–516, 1977
Diegelmann RF, Peterkofsky B: Collagen biosynthesis during connective tissue development in chick embryo. Dev Biol 28: 443–453, 1972
Rennard SI, Berg R, Martin GR, Foidart JM, Robey PG: Enzyme-linked immunoassay (ELISA) for connective tissue components. Anal Biochem 104: 205–214, 1980
Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ: Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18: 5294–5299, 1979
Sambrook J, Fritsch EF, Maniatis T: Extraction, purification and analysis of messenger RNA from eukaryotic cells. In: J. Sambrook, E.F. Fritsch and T. Maniatis (eds). Molecular cloning: a laboratory manual. 2nd Ed., Vol. 1. Cold Spring Harbor Laboratory Press, Plainview, NY, 1989, pp 7.01–7.87
Genovese C, Rowe D, Kream B: Construction of DNA sequences complementary to rat α1 and α2 collagen mRNA and their use in studying the regulation of type I collagen synthesis by 1,25-Dihydroxy-vitamin D. Biochemistry 23: 6210–6216, 1984
Tyagi SC, Matsubara L, Weber KT: Direct extraction and estimation of collagenase(s) activity by zymography in microquantities of rat myocardium and uterus. Clin Biochem 26: 191–198, 1993
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970
Goldblatt H: Experimental hypertension induced by renal ischemia (Harvey Lecture). Bull NY Acad Med 14: 523–553, 1938
Selye H: The general adaptation syndrome and the diseases of adaptation. J Clin Endocrinol 6: 117–230, 1946
Hall CE, Hall O: Hypertension and hypersalimentation. I. Aldosterone hypertension. Lab Invest 14: 285–294, 1965
Sun Y, Ratajska A, Zhou G, Weber KT: Angiotensin converting enzyme and myocardial fibrosis in the rat receiving angiotensin II or aldosterone. J Lab Clin Med 122: 395–403, 1993
Brilla CG, Pick R, Tan LB, Janicki JS, Weber KT: Remodeling of the rat right and left ventricle in experimental hypertension. Circ Res 67: 1355–1364, 1990
Lee AA, Dillman WH, McCulloch AD, Villarreal FJ: Angiotensin II stimulates the autocrine production of transforming growth factor-β1 in adult rat cardiac fibroblasts. J Mol Cell Cardiol 27: 2347–2357, 1995
Schelling P, Fischer H, Ganten D: Angiotensin and cell growth: a link to cardiovascular hypertrophy? J Hypertens 9: 3–15, 1991
Naftilan AJ, Pratt RE, Dzau VJ: Induction of platelet-derived growth factor A-chain and c-myc gene expressions by angiotensin II in cultured rat vascular smooth muscle cells. J Clin Invest 83: 1419–1424, 1989
Campbell SE, Katwa LC, Weber KT: Stimulation of transforming growth factor-beta by angiotensin II and/or aldosterone in cultured cardiac fibroblasts and valvular interstitial cells [Abstract]. Clin Res 4: 364A, 1994
Nagasawa K, Zimmermann R, Westernacher D, Schaper J: Angiotensin II induced collagen mRNA expression in cardiac fibroblasts depends on autocrine production of transforming growth factor-beta 1 [Abstract]. J Am Coll Cardiol 24, Suppl A: 43A, 1994
Lombès M, Oblin M-E, Gasc J-M, Baulieu EE, Farman N, Bonvalet J-P: Immunohistochemical and biochemical evidence for a cardiovascular mineralocorticoid receptor. Circ Res 71: 503–510, 1992
Slight S, Ganjam VK, Nonneman DJ, Weber KT: Glucocorticoid metabolism in the cardiac interstitium: 11β-hydroxysteroid dehydrogenase activity in cardiac fibroblasts. J Lab Clin Med 122: 180–187, 1993
Tyagi SC, Ratajska A, Weber KT: Myocardial matrix metalloproteinase(s): localization and activation. Mol Cell Biochem 126: 49–59, 1993
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This work was supported in part by NIH grant #RO1-HL-31701.
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Zhou, G., Kandala, J.C., Tyagi, S.C. et al. Effects of angiotensin II and aldosterone on collagen gene expression and protein turnover in cardiac fibroblasts. Mol Cell Biochem 154, 171–178 (1996). https://doi.org/10.1007/BF00226785
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DOI: https://doi.org/10.1007/BF00226785