Skip to main content
SpringerLink
Log in

Use of Combined Chemotherapy with Etoposide and Methotrexate, both Associated to Lipid Nanoemulsions for Atherosclerosis Treatment in Cholesterol-fed Rabbits

  • ORIGINAL ARTICLE
  • Published:
Cardiovascular Drugs and Therapy Aims and scope Submit manuscript

Abstract

Purpose

Treatment of atherosclerotic rabbits with intravenous methotrexate or etoposide carried in lipid nanoemulsions (LDE) markedly reduced the lesions in the aorta. Here, the combined treatment with LDE-methotrexate and LDE-etoposide was investigated aiming to increase the anti-atherosclerosis effect.

Methods

Thirty-six male rabbits received a diet with 1 % cholesterol for 2 months. After the first month, the animals received 4 weekly intravenous injections of LDE-methotrexate (4 mg/kg dose), LDE-etoposide (6 mg/kg), or a combination of those two drugs, while the control animals were injected with LDE (n = 9 for each group).

Results

LDE-methotrexate+LDE-etoposide reduced aortic lesion areas by 95 % compared with controls and the intima-media ratio was reduced five-fold, whereas LDE-methotrexate reduced the lesions by 81 % and LDE-etoposide by 83 %. Compared to controls, the positive area of macrophages and MMP-9 in the arterial intima was significantly reduced in all treated groups (p < 0.001), but the MMP9 reduction was greater with the combined chemotherapy than the reduction achieved by the isolated treatments. Presence of CD3 positive cells was equal in controls and LDE-methotrexate+LDE-etoposide treated animals. However, FOXP3 positive T lymphocytes in the intima were increased in the LDE-methotrexate+LDE-etoposide rabbits. Weight, food intake evolution and the hematologic parameters suggested that the treatment had very low toxicity.

Conclusions

Compared to the single treatments with LDE-methotrexate and LDE-etoposide, the combined treatment was more effective in reducing the atherosclerotic lesions. Because the toxicity of the novel drug-target combined scheme was low, those results favor the possibility of future clinical studies in patients with cardiovascular disease.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Charo IF, Taub R. Anti-inflammatory therapeutics for the treatment of atherosclerosis. Nat Rev Drug Discov. 2011;10:365–76.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  2. Ross R. Atherosclerosis - an inflammatory disease. N Engl J Med. 1999;14:115–26.

    Google Scholar 

  3. Maranhão RC, Roland IA, Toffoletto O, Ramires JA, Gonçalves RP, Mesquita CH. Plasma kinetic behavior in hyperlipidemic subjects of a lipidic microemulsion that binds to low density lipoprotein receptors. Lipids. 1997;32:627–33.

    Article  PubMed  Google Scholar 

  4. Maranhao RC, Tavares ER, Padoveze AF, Valduga CJ, Rodrigues DG, Pereira MD. Paclitaxel associated with cholesterol-rich nanoemulsions promotes atherosclerosis regression in the rabbit. Atherosclerosis. 2008;197:959–66.

    Article  CAS  PubMed  Google Scholar 

  5. Tavares ER, Freitas FR, Diament J, Maranhão RC. Reduction of atherosclerotic lesions in rabbits treated with etoposide associated with cholesterol-rich nanoemulsions. Int J Nanomedicine. 2011;6:2297–304.

    PubMed Central  CAS  PubMed  Google Scholar 

  6. Bulgarelli AC, Leite Jr AC, Dias AA, Maranhão RC. Anti-atherogenic effects of methotrexate carried by a lipid nanoemulsion that binds to LDL receptors in cholesterol-fed rabbits. Cardiovasc Drugs Ther. 2013;27:531–9.

    Article  CAS  PubMed  Google Scholar 

  7. Valduga CJ, Fernandes DC, Lo Prete AC, Azevedo CHM, Rodrigues DG, Maranhao RC. Use of a cholesterol-rich microemulsion that binds to low-density lipoprotein receptors as vehicle for etoposide. J Pharm Pharmacol. 2003;55:1615–22.

    Article  CAS  PubMed  Google Scholar 

  8. Dias ML, Carvalho JP, Rodrigues DG, Graziani SR, Maranhão RC. Pharmacokinetics and tumor uptake of a derivatized form of paclitaxel associated to a cholesterol rich nanoemulsion in patients with gynecologic cancers. Cancer Chemother Pharmacol. 2007;59:105–11.

    Article  CAS  PubMed  Google Scholar 

  9. Maranhão RC, Graziani SR, Yamaguchi N, Melo RF, Latrilha MC, Rodrigues DG, et al. Association of carmustine with a lipid emulsion: in vitro, in vivo and preliminary studies in cancer patients. Cancer Chemother Pharmacol. 2002;49:487–98.

    Article  PubMed  Google Scholar 

  10. Ginsburg GS, Small DM, Atkinson D. Microemulsion of phospholipids and cholesterol esters Protein-free models of low density lipoprotein. J Biol Chem. 1982;257:8216–27.

    CAS  PubMed  Google Scholar 

  11. Maranhão RC, César TB, Pedroso-Mariani SR, Hirata MH, Mesquita CH. Metabolic behavior in rats of a nonprotein microemulsion resembling low density lipoprotein. Lipids. 1993;28:691–6.

    Article  PubMed  Google Scholar 

  12. Rosowsky A, Forsch RA, Yu CS, Lazarus H, Beardsley GP. Methotrexate analogues. Divergent influence of alkyl chain on the dihydrofolate reductase affinity and cytotoxicity of methotrexate monoesters. J Med Chem. 1984;27:605–9.

    Article  CAS  PubMed  Google Scholar 

  13. Hall TC, Roberts D, Kessel DH. Methotrexate and folic reductase in human cancer. Eur J Cancer. 1996;2:135–42.

    Article  Google Scholar 

  14. Bonney RJ, Maley F. Effect of methotrexate on thymidylate synthetase in cultured parenchymal cells isolated from regenerating rat liver. Cancer Res. 1975;8:1950–6.

    Google Scholar 

  15. Engel R, Valkov NI, Gump JL, Hazlehurst L, Dalton WS, Sullivan DM. The cytoplasmic trafficking of DNA topoisomerase II alpha correlates with etoposide resistance in human myeloma cells. Exp Cell Res. 2004;295:421–31.

    Article  CAS  PubMed  Google Scholar 

  16. Wessels JA, Huizinga TW, Guchelaar HJ. Recent insights in the pharmacological actions of methotrexate in the treatment of rheumatoid arthritis. Rheumatology (Oxford). 2008;47:249–55.

    Article  CAS  Google Scholar 

  17. Tian H, Cronstein BN. Understanding the mechanisms of action of methotrexate: implications for the treatment of rheumatoid arthritis. Bull NYU Hosp Jt Dis. 2007;65:168–73.

    PubMed  Google Scholar 

  18. Bulgarelli A, Martins Dias AA, Caramelli B, Maranhão RC. Treatment with methotrexate inhibits atherogenesis in cholesterol-fed rabbits. J Cardiovasc Pharmacol. 2012;59:308–14.

    Article  CAS  PubMed  Google Scholar 

  19. de la Llera-Moya M, Rothblat GH, Glick JM, England JM. Etoposide treatment suppresses atherosclerotic plaque development in cholesterol-fed rabbits. Arterioscler Thromb. 1992;12:1363–70.

    Article  PubMed  Google Scholar 

  20. Esparza L, De Haro J, Bleda S, Acin F. Non-Fas (CD95/APO1)-mediated apoptosis of activated T cells inhibits thedevelopment of atherosclerosis. Interact Cardiovasc Thorac Surg. 2012;15:340–3.

    Article  PubMed Central  PubMed  Google Scholar 

  21. Galis ZS, Khatri JJ. Matrix metalloproteinases in vascular remodeling and atherogenesis: the good, the bad, and the ugly. Circ Res. 2002;3:251–62.

    Google Scholar 

  22. Jonasson L, Holm J, Skalli O, Bondjers G, Hansson GK. Regional accumulations of T cells, macrophages, and smooth muscle cells in the human atherosclerotic plaque. Arteriosclerosis. 1996;6:131–8.

    Article  Google Scholar 

  23. de Boer OJ, van der Meer JJ, Teeling P, van der Loos CM, van der Wal AC. Low numbers of foxp3 positive regulatory t cells are present in all developmental stages of human atherosclerotic lesions. PLoS One. 2007;2(8):e779.

    Article  PubMed Central  PubMed  Google Scholar 

  24. Ridker PM. Testing the inflammatory hypothesis of atherothrombosis: scientific rationale for the cardiovascular inflammation reduction trial (CIRT). J Thromb Haemost. 2009;7 Suppl 1:332–9.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was financially supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), São Paulo, and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Brasília, Brazil. Dr Maranhão has a Research Award from CNPq. We thank Maria das Dores for technical support.

Conflict of Interest

The authors report no conflicts of interest in this work.

Author information

Authors and Affiliations

  1. Lipid Metabolism Laboratory, Heart Institute (INCOR) and the Medical School Hospital, São Paulo, Av. Dr. Eneas C. Aguiar 44, 1 subsolo, São Paulo, SP, CEP 0543000, Brazil

    Antonio C. A. Leite Jr., Tatiana V. Solano, Elaine R. Tavares & Raul C. Maranhão

  2. Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, SP, Brazil

    Antonio C. A. Leite Jr., Tatiana V. Solano & Raul C. Maranhão

Authors
  1. Antonio C. A. Leite Jr.
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tatiana V. Solano
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Elaine R. Tavares
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Raul C. Maranhão
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Raul C. Maranhão.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Leite, A.C.A., Solano, T.V., Tavares, E.R. et al. Use of Combined Chemotherapy with Etoposide and Methotrexate, both Associated to Lipid Nanoemulsions for Atherosclerosis Treatment in Cholesterol-fed Rabbits. Cardiovasc Drugs Ther 29, 15–22 (2015). https://doi.org/10.1007/s10557-014-6566-1

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10557-014-6566-1

Keywords

Search

Navigation