Molecular and Cellular Biochemistry

, Volume 136, Issue 2, pp 97–103 | Cite as

Nicotine regulates collagen gene expression, collagenase activity, and DNA synthesis in cultured cardiac fibroblasts

  • Ronald J. Tomek
  • Stephen Rimar
  • Mahboubeh Eghbali-Webb


Cardiac fibroblasts that reside in the interstitium are the cellular origin of collagen and other proteins of the extracellular matrix in the heart. We have previously shown thatin vitro gene expression, proliferation and even phenotypic features of cardiac fibroblasts are subject to regulation by biological factors such as hormones, growth factors and neurotransmitters. The influence of nicotine, the active ingredient of tobacco, on risk factors for cardiac diseases is well known.In vivo adverse effects of nicotine are as the result of its direct and indirect effects. The cellular and molecular mechanisms of direct effects of nicotine in the heart are widely unknown. The objective of this study was to investigate if nicotine has direct influence on cardiac fibroblasts. To this end, we studied the effects of nicotine on cultured cardiac fibroblasts. Northern hybridization analysis of RNA extracted from cardiac fibroblasts, enzymography of conditioned medium of cardiac fibroblasts and [3H]-thymidine incorporation into DNA of cardiac fibroblasts were used to examine the effects of nicotine on collagen gene expression, collagenase activity and DNA synthesis respectively. Treatment of cardiac fibroblasts with nicotine (10 μg/ml) led to a 31% (P<0.05) decrease in the abundance of mRNA for pro α1(I) but not pro α2(I) collagen compared with control untreated cells. Nicotine treatment of cardiac fibroblasts also led to decreased collagenase activity (62%, P<0.001) in the conditioned medium of those cells in culture. Studies with [3H]-thymidine incorporation into DNA of cardiac fibroblasts showed a nicotine-induced decrease (39%, P<0.001) in DNA synthesis in those cells. These findings suggest that cardiac fibroblasts are targets for the toxic effects of nicotine. The findings further point to the possibility that nicotine-induced alterations in cardiac fibroblasts' function and gene expression may contribute to the biological processes that ultimately lead to adverse effects of nicotine in the heart.

Key words

fibroblasts collagen metabolism heart smoking DNA synthesis extracellular matrix tobacco 


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  1. 1.
    Kuller LH, Ockene JK, Meilahn K, Wentworth D, Svendsen KH, Neaton JD: Cigarette smoking and mortality. Prev Med 20: 638–654, 1991Google Scholar
  2. 2.
    Richard JL, Ducimetiére P, Bonnaud G, Claud JR, Lellouch J, Schwartz D, Dimatteo J: Incidence et evaluation du risque de maladie coronarienne. Arch Mal Coeur Vaiss 70: 531–540, 1977Google Scholar
  3. 3.
    Ramp WK, Lenz LG, Galvin RJS: Nicotine inhibits collagen synthesis and alkaline phosphatase activity, but stimulates DNA synthesis in osteoblast-like cells. PSEBM 197: 36–43, 1991Google Scholar
  4. 4.
    Chamson, A, Frey J: Effects of tobacco smoke extracts on collagen biosynthesis by fibroblast cell cultures. J Toxicol Env Health 9: 921–937, 1982Google Scholar
  5. 5.
    Hanes JP, Schuster GS, Lubas S: Binding, uptake and release of nicotine by human gingival fibroblasts. J Periodontal 62: 147–152, 1991Google Scholar
  6. 6.
    Schuller HM: Cell type-specific receptor-mediated modulation of growth kinetics in human lung cancer cell lines by nicotine and tobacco-related nitrosamines. Pharmacology 38: 3439–3442, 1989Google Scholar
  7. 7.
    Eghbali M, Czaja MJ, Zeydel M, Weiner FR, Zern MA, Seifter S, Blumenfeld OO: Collagen mRNAs in isolated adult heart cell. J Mol Cell Cardiol 20: 267–276, 1988Google Scholar
  8. 8.
    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, 1989Google Scholar
  9. 9.
    Eghbali M, Weber KT: Collagen and the myocardium: Fibrillar structure, biosynthesis and degradation in relation to hypertrophy and its regression. Mol Cell Biochem 96: 1–14, 1990Google Scholar
  10. 10.
    Eghbali M, Tomek R, Sukhatme V, Woods C, Bhambi B: Differential effects of transforming growth factor-β1 and phorbol mirystate acetate on cardiac fibroblasts: Regulation of fibrillar collagen mRNAs and expression of early transcription factors. Circ Res 69: 483–490, 1991Google Scholar
  11. 11.
    Eghbali M, Tomek R, Woods C, Bhambi B: Cardiac fibroblasts are predisposed to convert into myocyte phenotype: Specific effects of transforming growth factor-beta. Proc Natl Acad Sci USA 88: 795–799, 1991Google Scholar
  12. 12.
    Bhambi B, Eghbali M: Effect of norepinephrine on myocardial collagen gene expression and response of cardiac fibroblasts following norepinephrine treatment. Am J Pathol 139: 1131–1142, 1991Google Scholar
  13. 13.
    Yao J, Eghbali M: Decreased collagen gene expression and absence of fibrosis in thyroid hormone-induced myocardial hypertrophy: Response of cardiac fibroblasts to thyroid hormone in vitro. Circ Res 71: 831–839, 1992Google Scholar
  14. 14.
    Benowitz NL, Jaacob PIII, Yu L: Daily use of smokeless tobacco: systemic effects. Ann Intern Med III: 112–116, 1989Google Scholar
  15. 15.
    Chomezynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocynate-phenol-chloroform extraction. Anal Biochem 162: 156–159, 1987Google Scholar
  16. 16.
    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-Dihydroxyvitamin D. Biochemistry 23: 6210–6216, 1984Google Scholar
  17. 17.
    Cleveland DW, Lopata MA, MacDonald RJ, Cowan NJ, Rutter WJ, Kirschner MW: Number and evolutionary conservation of α- and β-tubulin and cytoplasmic β- and α-actin genes using specific cloned cDNA probes. Cell 20: 95–105, 1980Google Scholar
  18. 18.
    Derynck R, Rhee L: Sequence of the porcine transforming growth factor β. Nucleic Acid Res 15:3187, 1987Google Scholar
  19. 19.
    Feinberg AP, Vogelstein B: A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13, 1983Google Scholar
  20. 20.
    Brenner CA, Adler RR, Rappolee DA, Pedersen RA, Werb Z: Genes for extracellular matrix-degrading metaloproteinases and their inhibitor TIMP, are expressed during early mammalian development. Genes and Develop 3: 848–859, 1989Google Scholar
  21. 21.
    Correra-Rotter R, Mariash CN, Rosenberg ME: Loading and transfer control for Northern hybridization. Biotechniques 12: 154–158, 1992Google Scholar
  22. 22.
    Smith BD, Niles R: Characterization of collagen synthesized by normal and chemically transformed rat liver epithelial cell lines. Biochemistry 19: 1820–1825, 1989Google Scholar
  23. 23.
    Sandmeyer S, Smith R, Kiehn D, Bornstein P: Correlation of collagen synthesis and procollagen messenger RNA levels with transformation in rat embryo fibroblasts. Cancer Res 41: 830–838, 1981Google Scholar
  24. 24.
    Neher GH: Nicotine-induced depression of lymphocyte growth. Toxicol Appl Pharmacol 27: 253–258, 1974Google Scholar

Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • Ronald J. Tomek
    • 1
  • Stephen Rimar
    • 1
  • Mahboubeh Eghbali-Webb
    • 1
  1. 1.Department of AnesthesiologyYale University School of MedicineNew HavenUSA

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