Skip to main content
SpringerLink
Log in

Role of COX-2 in the Bioactivation of Methylenedianiline and in Its Proliferative Effects in Vascular Smooth Muscle Cells

Cardiovascular Toxicology volume 11, pages 316–324 (2011)Cite this article

Abstract

4,4′-Methylenedianiline (DAPM) is an aromatic diamine used directly in the production of polyurethane foams and epoxy resins, or as a precursor to MDI in the manufacture of some polyurethanes. In our prior experiments, we showed that chronic, intermittent treatment of female rats with DAPM resulted in vascular medial hyperplasia of pulmonary arteries. In addition, treatment of vascular smooth muscle cells (VSMC) in culture with DAPM increased the rates of proliferation in a manner that was inhibited by co-treatment with N-acetylcysteine but was not associated with oxidative stress. We thus hypothesized that NAC treatment inhibited DAPM toxicity by competing for binding reactive intermediates formed through DAPM metabolism. Because the peroxidase enzyme cyclooxygenase is constitutively expressed in VSMC, and because cyclooxygenase is known to metabolize similar aromatic amines to electrophilic intermediates, we further hypothesized that DAPM-induced VSMC proliferation was dependent upon COX-1/2-mediated bioactivation. To test this hypothesis, we treated VSMC with DAPM and measured cell proliferation, COX-2 expression, COX-1/2 activity, and levels of covalent binding. DAPM treatment resulted in a dose-dependent increase in proliferation that was abolished by co-treatment with the COX-2-selective inhibitor celecoxib. In addition, DAPM exposure increased the rates of proliferation in VSMC isolated from wild-type but not COX-2 (−/−) mice. Paradoxically, treatment with DAPM reduced the cellular production of PGE2 and PGF2α, but dose-dependently increased the COX-2 protein levels. Covalent binding of [14C]-DAPM to VSMC biomolecules was greater in wild-type than in COX-2 (−/−) cells. However, covalent binding of [14C]-DAPM was not altered by co-treatment with a nonselective inhibitor of cytochromes P450. These studies thus suggest that DAPM-induced VSMC proliferation may be due to bioactivation of DAPM, perhaps through the action of cyclooxygenase. The data furthermore suggest that DAPM’s mechanism of action may possibly involve inhibition or suicide inactivation of COX-2. In addition, because we observed an increase in DAPM-induced VSMC proliferation in cells isolated from female compared to male rats, further studies into the potential interplay between DAPM, the estrogen receptor, and COX-2 seem warranted.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

39,95 €

Price includes VAT (Finland)

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

References

  1. A Public Health Statement for 4,4′-Methylenedianiline. (1998, August). Agency for Toxic Substances and Disease Registry. Retrieved June, 2011 from http://www.atsdr.cdc.gov/phs/phs.asp?id=999&tid=210.

  2. Preambles to Final Rules, Section 8—VIII. (1992, August). Summary of the regulatory Impact analysis and regulatory flexibility analysis. Regulations: OSHA.

  3. Kanz, M. F., Gunasena, G. H., Kaphalia, L., Hammond, D. K., & Syed, Y. A. (1998). A minimally toxic dose of methylene dianiline injures biliary epithelial cells in rats. Toxicology and Applied Pharmacology, 150, 414–426.

    Article  PubMed  CAS  Google Scholar 

  4. Cruz, V. S., Dugas, T. R., & Kanz, M. F. (2005). Mitochondrial dysfunction occurs before transport or tight junction deficits in biliary epithelial cells exposed to bile from methylenedianiline-treated rats. Toxicological Sciences, 84, 129–138.

    Article  Google Scholar 

  5. Dugas, T. R., Cruz, V. S., Liu, H., & Kanz, M. F. (2001). Evaluation of the gender differences in 4, 4′-methylenedianiline toxicity, distribution, and effects on biliary parameters. Journal of Toxicology and Environmental Health Part A, 62, 467–483.

    Article  PubMed  CAS  Google Scholar 

  6. Dugas, T. R., Kanz, M. F., Hebert, V. Y., Hennard, K. L., Liu, H., Cruz, V. S., et al. (2004). Vascular medial hyperplasia following chronic, intermittent exposure to 4, 4′-methylenedianiline. Cardiovascular Toxicology, 4, 85–96.

    Article  PubMed  CAS  Google Scholar 

  7. Eling, T. E., Thompson, D. C., Foureman, G. L., Curtis, J. F., & Hughes, M. F. (1990). Prostaglandin H synthase and xenobiotic oxidation. Annual Review of Pharmacology and Toxicology, 30, 1–45.

    Article  PubMed  CAS  Google Scholar 

  8. Chen, K., Dugas, T. R., & Cole, R. B. (2006). Identification of metabolites of 4, 4′-methylenedianiline in vascular smooth muscle cells by liquid chromatography-electrospray tandem mass spectrometry. Journal of Mass Spectrometry, 41, 728–734.

    Article  PubMed  CAS  Google Scholar 

  9. Yan, Z., Subbaramaiah, K., Camilli, T., Zhang, F., Tanabe, T., McCaffrey, T. A., et al. (2000). Benzo[a]pyrene induces the transcription of cyclooxygenase-2 in vascular smooth muscle cells. Evidence for the involvement of extracellular signal-regulated kinase and NF-kappaB. Journal of Biological Chemistry, 275, 4949–4955.

    Article  PubMed  CAS  Google Scholar 

  10. Mitchell, J. A., Larkin, S., & Williams, T. J. (1995). Cyclooxygenase-2: Regulation and relevance in inflammation. Biochemical Pharmacology, 50, 1535–1542.

    Article  PubMed  CAS  Google Scholar 

  11. Burleigh, M. E., Babaev, V. R., Yancey, P. G., Major, A. S., McCaleb, J. L., Oates, J. A., et al. (2005). Cyclooxygenase-2 promotes early atherosclerotic lesion formation in ApoE-deficient and C57BL/6 mice. Journal of Molecular and Cellular Cardiology, 39, 443–452.

    Article  PubMed  CAS  Google Scholar 

  12. Connolly, E., Bouchier-Hayes, D. J., Kaye, E., Leahy, A., Fitzgerald, D., & Belton, O. (2002). Cyclooxygenase isozyme expression and intimal hyperplasia in a rat model of balloon angioplasty. The Journal of Pharmacology and Experimental Therapeutics, 300, 393–398.

    Article  PubMed  CAS  Google Scholar 

  13. Vachiery, J. L. (2011). Prostacyclins in pulmonary arterial hypertension: The need for earlier therapy. Advances in Therapy, 28, 251–269.

    Article  PubMed  CAS  Google Scholar 

  14. Uehara, Y., Takada, S., Hirawa, N., Kawabata, Y., Nagata, T., Numabe, A., et al. (1993). De novo synthesis of phospholipase A2 and prostacyclin production by proliferating rat smooth muscle cells. Prostaglandins, 46, 331–346.

    Article  PubMed  CAS  Google Scholar 

  15. Libby, P., Warner, S. J., & Friedman, G. B. (1988). Interleukin 1: A mitogen for human vascular smooth muscle cells that induces the release of growth-inhibitory prostanoids. The Journal of Clinical Investigation, 81, 487–498.

    Article  PubMed  CAS  Google Scholar 

  16. Otto, A. M., Nilsen-Hamilton, M., Boss, B. D., Ulrich, M. O., & Jimenez De Asua, L. (1982). Prostaglandins E1 and E2 interact with prostaglandin F2alpha to regulate initiation of DNA replication and cell division in swiss 3T3 cells. Proceedings of the National Academy of Sciences of the United States of America, 79, 4992–4996.

    Article  PubMed  CAS  Google Scholar 

  17. Hara, S., Morishita, R., Tone, Y., Yokoyama, C., Inoue, H., Kaneda, Y., et al. (1995). Overexpression of prostacyclin synthase inhibits growth of vascular smooth muscle cells. Biochemical and Biophysical Research Communications, 216, 862–867.

    Article  PubMed  CAS  Google Scholar 

  18. Sachinidis, A., Flesch, M., Ko, Y., Schror, K., Bohm, M., Dusing, R., et al. (1995). Thromboxane A2 and vascular smooth muscle cell proliferation. Hypertension, 26, 771–780.

    PubMed  CAS  Google Scholar 

  19. Bishop-Bailey, D., Hla, T., & Mitchell, J. A. (1999). Cyclo-oxygenase-2 in vascular smooth muscle. International Journal of Molecular Medicine, 3, 41–48.

    PubMed  CAS  Google Scholar 

  20. Fredenburgh, L. E., Ma, J., & Perrella, M. A. (2009). Cyclooxygenase-2 inhibition and hypoxia-induced pulmonary hypertension: Effects on pulmonary vascular remodeling and contractility. Trends in Cardiovascular Medicine, 19, 31–37.

    Article  PubMed  CAS  Google Scholar 

  21. Choi, H. C., Kim, H. S., Lee, K. Y., Chang, K. C., & Kang, Y. J. (2008). NS-398, a selective COX-2 inhibitor, inhibits proliferation of IL-1beta-stimulated vascular smooth muscle cells by induction of HO-1. Biochemical and Biophysical Research Communications, 376, 753–757.

    Article  PubMed  CAS  Google Scholar 

  22. Fosslien, E. (2005). Cardiovascular complications of non-steroidal anti-inflammatory drugs. Annals of Clinical and Laboratory Science, 35, 347–385.

    PubMed  CAS  Google Scholar 

  23. Fries, S., & Grosser, T. (2005). The cardiovascular pharmacology of COX-2 inhibition. Hematology/the Education Program of the American Society of Hematology, 2005, 445–451.

    Article  Google Scholar 

  24. Cyrus, T., Sung, S., Zhao, L., Funk, C. D., Tang, S., & Pratico, D. (2002). Effect of low-dose aspirin on vascular inflammation, plaque stability, and atherogenesis in low-density lipoprotein receptor-deficient mice. Circulation, 106, 1282–1287.

    Article  PubMed  CAS  Google Scholar 

  25. Speir, E., Yu, Z. X., Ferrans, V. J., Huang, E. S., & Epstein, S. E. (1998). Aspirin attenuates cytomegalovirus infectivity and gene expression mediated by cyclooxygenase-2 in coronary artery smooth muscle cells. Circulation Research, 83, 210–216.

    PubMed  CAS  Google Scholar 

  26. Pratico, D., Tillmann, C., Zhang, Z. B., Li, H., & FitzGerald, G. A. (2001). Acceleration of atherogenesis by COX-1-dependent prostanoid formation in low density lipoprotein receptor knockout mice. Proceedings of the National Academy of Sciences of the United States of America, 98, 3358–3363.

    Article  PubMed  CAS  Google Scholar 

  27. Kautiainen, A., Wachtmeister, C. A., & Ehrenberg, L. (1998). Characterization of hemoglobin adducts from a 4, 4′-methylene dianiline metabolite evidently produced by peroxidative oxidation in vivo. Chemical Research in Toxicology, 11, 614–621.

    Article  PubMed  CAS  Google Scholar 

  28. Josephy, P. D. (1985). Oxidative activation of benzidine and its derivatives by peroxidases. Environmental Health Perspectives, 64, 171–178.

    Article  PubMed  CAS  Google Scholar 

  29. Yamakawa, T., Ohnaka, K., Tanaka, S., Utsunomiya, H., Kamei, J., & Kadonosono, K. (2008). Cyclooxygenase-2 induction by lysophosphatidylcholine in cultured rat vascular smooth muscle cells: Involvement of the p38MAPK pathway. Biomedical Research, 29, 1–8.

    Article  PubMed  CAS  Google Scholar 

  30. Samuelsson, B., Morgenstern, R., & Jakobsson, P. J. (2007). Membrane prostaglandin E synthase-1: A novel therapeutic target. Pharmacological Reviews, 59, 207–224.

    Article  PubMed  CAS  Google Scholar 

  31. Chang, W. C., Nakao, J., Neichi, T., Orimo, H., & Murota, S. (1981). Effects of estradiol on the metabolism of arachidonic acid by aortas and platelets in rats. Biochimica et Biophysica Acta, 664, 291–297.

    PubMed  CAS  Google Scholar 

  32. Doualla-Bell, F., Guay, J. M., Bourgoin, S., & Fortier, M. A. (1998). Prostaglandin G/H synthase (PGHS)-2 expression in bovine myometrium: Influence of steroid hormones and PGHS inhibitors. Biology of Reproduction, 59, 1433–1438.

    Article  PubMed  CAS  Google Scholar 

  33. Akarasereenont, P., Techatraisak, K., Thaworn, A., & Chotewuttakorn, S. (2000). The induction of cyclooxygenase-2 by 17beta-estradiol in endothelial cells is mediated through protein kinase C. Inflammation Research, 49, 460–465.

    Article  PubMed  CAS  Google Scholar 

  34. Ospina, J. A., Krause, D. N., & Duckles, S. P. (2002). 17beta-estradiol increases rat cerebrovascular prostacyclin synthesis by elevating cyclooxygenase-1 and prostacyclin synthase. Stroke, 33, 600–605.

    Article  PubMed  CAS  Google Scholar 

  35. Geary, G. G., McNeill, A. M., Ospina, J. A., Krause, D. N., Korach, K. S., & Duckles, S. P. (2001). Selected contribution: Cerebrovascular nos and cyclooxygenase are unaffected by estrogen in mice lacking estrogen receptor-alpha. Journal of Applied Physiology, 91, 2391–2399, discussion 2389–90.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pharmacology, Toxicology and Neuroscience, Louisiana State University Health Sciences Center, Shreveport, 1501 Kings Highway, Shreveport, LA, 71103, USA

    Valeria Y. Hebert, Brandon Chad Jones, Randy C. Mifflin & Tammy R. Dugas

Authors
  1. Valeria Y. Hebert
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Brandon Chad Jones
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Randy C. Mifflin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tammy R. Dugas
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tammy R. Dugas.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Hebert, V.Y., Jones, B.C., Mifflin, R.C. et al. Role of COX-2 in the Bioactivation of Methylenedianiline and in Its Proliferative Effects in Vascular Smooth Muscle Cells. Cardiovasc Toxicol 11, 316–324 (2011). https://doi.org/10.1007/s12012-011-9123-1

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12012-011-9123-1

Keywords

  • Vascular smooth muscle cell proliferation
  • Aromatic amine
  • Bioactivation
  • Cyclooxygenase
  • Peroxidase
  • Metabolism

Search

Navigation