Document Type: Research(Original) Article

Authors

1 Department of Research Center, Gerash Faculty of Medical Sciences, Shiraz University of medical Sciences, Shiraz, Iran

2 Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.

3 1. Cellular and Molecular Gerash Research Center, Shiraz University of Medical Sciences, Shiraz, Iran. 2. Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.

Abstract

Introduction: Cyclophosphamide is a chemo-therapeutic agent in the treatment of the various cancers and autoimmune diseases. This composition has cytotoxic and clastogenic properties. The purpose of this study was to evaluate, protective effect of methanol extracts of thymus vulgaris against DNA damage induced by cyclophosphamide in mouse bone marrow cells by the micronucleus test.Materials and Methods: In this study, the concentrations of extracts 375, 750, 1500 mg/kg body weight (bw) were injected intraperitoneally (ip), into mice for seven consecutive days. One hour after the last injection, cyclophosphamide 50 mg/kg bw intraperitoneally (ip) have been injected into mice. 24h after cyclophosphamide injection, the animals were sacrificed and samples of bone marrow were prepared and stained using standard methods. For each sample, 1000 cells of polychromatic erythrocytes (PCE) and the same frequence of normochromatic erythrocyte (NCE) and cells containing micronucleus of these were counted.Results: Cyclophosphamide increased the frequency of micronuclei polychromatic erythrocytes (MnPCE) and decreased cell proliferation (PCE/PCE+NCE). All doses of extracts significantly reduced the micronucleus frequency ratio (P<0.05). The cells proliferation ratio (PCE/PCE+NCE) also increased. The best effect in reducing the micronucleus frequency was 1500 mg/kg bw dosage.Conclusion: Thymus extract is able to reduce the clastogenic and cytotoxic effects of cyclophosphamide, seems to extract due to its antioxidant properties, has a protective role.

  1. Flora SD, Izzotti A, D’Agostini F, Balansky RM, Noonan D, Albini A. Multiple points of intervention in the prevention of cancer and other mutation-related diseases. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis. 2001;480:9-22.
  2. Hales BF, Barton TS, Robaire B. Impact of paternal exposure to chemotherapy on offspring in the rat. JNCI Monographs. 2005;2005(34):28-31.
  3. Qiu J, Hales BF, Robaire B. Damage to rat spermatozoal DNA after chronic cyclophosphamide exposure. Biology of reproduction. 1995;53(6):1465-73.
  4. Kern JC, Kehrer JP. Acrolein-induced cell death: a caspase-influenced decision between apoptosis and oncosis/necrosis. Chemico-biological interactions. 2002;139(1):79-95.
  5. Bergamini CM, Gambetti S, Dondi A, Cervellati C. Oxygen, reactive oxygen species and tissue damage. Current pharmaceutical design. 2004;10(14):1611-26.
  6. Duthie GG, Duthie SJ, Kyle JA. Plant polyphenols in cancer and heart disease: implications as nutritional antioxidants. Nutrition Research Reviews. 2000;13(1):79-106.
  7. Jangjoo M. Collected, Naming Scientific and of Phytochemical Plants of the Fars Region Gerash: Tehran University of Medical Sciences; 1373.
  8. Zargari A. Medicinal plants: Tehran Univ.; 1997.
  9. Garcia-Risco MR, Vicente G, Reglero G, Fornari T. Fractionation of thyme (< i> Thymus vulgaris L.) by supercritical fluid extraction and chromatography. The Journal of Supercritical Fluids. 2011;55(3):949-54.
  10. Rota MC, Herrera A, Martinez RM, Sotomayor JA, Jordan MJ. Antimicrobial activity and chemical composition of Thymus vulgaris, Thymus zygis and Thymus hyemalis essential oils. Food Control. 2008;19(7):681-7.
  11. Kumar A, Shukla R, Singh P, Prasad CS, Dubey NK. Assessment of Thymus vulgaris L. essential oil as a safe botanical preservative against post harvest fungal infestation of food commodities. Innovative Food Science & Emerging Technologies. 2008;9(4):575-80.
  12. Nguefack J, Dongmo J, Dakole C, Leth V, Vismer H, Torp J, et al. Food preservative potential of essential oils and fractions from Cymbopogon citratus, Ocimum gratissimum and Thymus vulgaris against mycotoxigenic fungi. International journal of food microbiology. 2009;131(2):151-6.
  13. El-Nekeety AA, Mohamed SR, Hathout AS, Hassan NS, Aly SE, Abdel-Wahhab MA. Antioxidant properties of Thymus vulgaris oil against aflatoxin-induce oxidative stress in male rats. Toxicon. 2011;57(7):984-91.
  14. Taherian AA, Babaei M, Vafaei AA, Jarrahi M, Jadidi M, Sadeghi H. antinociceptive effects of hydroalcoholic extract of Thymus vulgaris. Pak J Pharm Sci. 2009;22(1):83-9.
  15. Schmid W. The micronucleus test. Mutation Research/Environmental Mutagenesis and Related Subjects. 1975;31(1):9-15.
  16. Müller W-U, Streffer C. Micronucleus assays. Advances in mutagenesis research: Springer; 1994. p. 1-134.
  17. Fenech M. The in vitro micronucleus technique. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis. 2000;455(1):81-95.
  18. Serpeloni JM, dos Reis MB, Rodrigues J, dos Santos LC, Vilegas W, Varanda EA, et al. In vivo assessment of DNA damage and protective effects of extracts from Miconia species using the comet assay and micronucleus test. Mutagenesis. 2008;23(6):501-7.
  19. Aguilar-Mahecha A, Hales BF, Robaire B. Chronic cyclophosphamide treatment alters the expression of stress response genes in rat male germ cells. Biology of reproduction. 2002;66(4):1024-32.
  20. Tiwari AK. Imbalance in antioxidant defence and human diseases: Multiple approach of natural antioxidants therapy. CURRENT SCIENCE-BANGALORE-. 2001;81(9):1179-87.
  21. Gamal-Eldeen AM, Abo-Zeid MA, Ahmed EF. Anti-genotoxic effect of the Sargassum dentifolium extracts: Prevention of chromosomal aberrations, micronuclei, and DNA fragmentation. Experimental and Toxicologic Pathology. 2013;65(1):27-34.
  22. Zhang QH, Wu CF, Yang JY, Mu YH, Chen XX, Zhao YQ. Reduction of cyclophosphamide-induced DNA damage and apoptosis effects of ginsenoside Rb1 on mouse bone marrow cells and peripheral blood leukocytes. Environmental Toxicology and Pharmacology. 2009;27(3):384-9.
  23. Vilar J, Leite K, Chen LC. Antimutagenicity protection of Ginkgo biloba extract (Egb 761) against mitomycin C and cyclophosphamide in mouse bone marrow. Genetics Molecular Res. 2009;8:328-33.
  24. Slamenova D, Horvathova E, Sramkova M, Marsalkova L. DNA-protective effects of two components of essential plant oils carvacrol and thymol on mammalian cells cultured in vitro. Neoplasma. 2007;54(2):108-12.
  25. Mastelic J, Jerkovic I, Blažević I, Poljak-Blaži M, Borović S, Ivančić-Baće I, et al. Comparative study on the antioxidant and biological activities of carvacrol, thymol, and eugenol derivatives. Journal of agricultural and food chemistry. 2008;56(11):3989-96.
  26. Samejima K, Kanazawa K, Ashida H, Danno G-i. Luteolin: a strong antimutagen against dietary carcinogen, Trp-P-2, in peppermint, sage, and thyme. Journal of agricultural and food chemistry. 1995;43(2):410-4.
  27. Ravizza R, Gariboldi MB, Molteni R, Monti E. Linalool, a plant-derived monoterpene alcohol, reverses doxorubicin resistance in human breast adenocarcinoma cells. Oncology reports. 2008;20(3):625-30.