Anti-Genotoxic Effect of Ocimum sanctum L. Extract against Cyproterone Acetate Induced Genotoxic Damage in Cultured Mammalian Cells

Abstract

The anti-genotoxic effect of Ocimum sanctum L. extract was studied against the genotoxic effect induced by a synthetic progestin cyproterone acetate, on human lymphocytes using chromosomal aberrations, mitotic index, sister chromatid exchanges and replication index as a parameters. About 30 μM of cyproterone acetate was treated with O. sanctum L. infusion, at dosages of 1.075 × 10−4, 2.125 × 10−4 and 3.15 × 10−4 g/ml of culture medium. A clear dose-dependent decrease in the genotoxic damage of cyproterone acetate was observed, suggesting a possible modulating role of the plant infusion. The results of the present study suggest that the plant infusion per se does not have genotoxic potential, but can modulate the genotoxicity of cyproterone acetate on human lymphocytes in vitro.

References

  1. 1.

    Agrawal, P., Rai, V., Singh, R. B. (1996) Randomized placebo controlled, single blind trial of holy basil leaves in patients with non-insulin dependent diabetes mellitus. Int. J. Clin. Pharmacol. Therap. 34, 406–409.

    CAS  Google Scholar 

  2. 2.

    Ahmad, S., Hoda, A., Afzal, M. (2002) Additive action of vitamins C and E against hydrocortisone induced genotoxicity in human lymphocytes chromosomes. Int. J. Vitam. Nutr. 72, 204–209.

    CAS  Google Scholar 

  3. 3.

    Aruna, K., Sivaramakrishnan, V. M. (1992) Anticarcinogenic effect of some Indian plant products. Food Chem. Toxicol. 30, 953–956.

    CAS  PubMed  Google Scholar 

  4. 4.

    Balanehru, S., Nagarajan, B. (1991) Protective effect of oleanolic acid and ursolic acid against lipid peroxidation. Biochem. Inter. 24, 981–990.

    CAS  Google Scholar 

  5. 5.

    Carballo, M. A., Alvarez, S., Boveris, S. (1993) Cellular stress by light and Rose Bengal in human lymphocytes. Mutat. Res. 288, 215–222.

    CAS  PubMed  Google Scholar 

  6. 6.

    Dastur, J. F. (1962) Medicinal Plants of India and Pakistan. D. B. Taraporevala Sons&Co. Bombay.

    Google Scholar 

  7. 7.

    Deml, E., Schwarz, L. R., Oesterle, D. (1993) Initiation of enzyme altered foci by the synthetic steroid cyproterone acetate in rat liver foci bioassay. Carcinogenesis 14, 1229–1231.

    CAS  PubMed  Google Scholar 

  8. 8.

    Devi, P. U., Bisht, K. S., Vinitha, M. (1998) A comparative study of radioprotection by flavonoids and synthetic aminothiol protectors in the mouse. British J. Radiol. 71, 782–784.

    CAS  Google Scholar 

  9. 9.

    Ganasoundari, A., Devi, P. U., Rao, B. S. S. (1998) Enhancement of bone marrow radioprotection and reduction of WR-2721 toxicity by Ocimum sanctum. Mutat. Res. 394, 303–312.

    Google Scholar 

  10. 10.

    Ganasoundari, A., Devi, P. U., Rao, M. N. A. (1997) Protection against radiation-induced chromosome damage in mouse bone marrow by Ocimum sanctum. Mutat. Res. 373, 271–276.

    CAS  PubMed  Google Scholar 

  11. 11.

    Ganasoundari, A., Zare, S. M., Devi, P. U. (1997) Modification of bone marrow radiosensitivity by medicinal plant extract. British J. Radiol. 70, 599.

    CAS  Google Scholar 

  12. 12.

    Gebhart, E. (1981) Sister chromatid exchange (SCE) and structural chromosome aberrations in mutagenicity testing. Hum. Genet. 58, 235–254.

    CAS  PubMed  Google Scholar 

  13. 13.

    Hayes, T. B., Stuart, A. A., Mendoza, M., Collins, A., Norieg, N., Vonk, A., Johmston., G., Liu, R., Kpodzo, D. (2006) Characterization of atrazine induced gonodal malformations in African clawed frogs (Xenopus laevis) and comparisons with effects of an androgen antagonist (cyproterone acetate) and exogenous estrogen (17 beta estradiol): support for the demasculization/feminization hypothesis. Environ. Health Perspect. 114, 134–141.

    PubMed  PubMed Central  Google Scholar 

  14. 14.

    IARC (1999) International Agency for Research on Cancer. Oral Contraceptive, Combined (Group 1) Vol. 72, Lyon, France, pp. 49.

    Google Scholar 

  15. 15.

    IARC (1999) Monographs on the hormonal contraceptives, progestogens only (Group 2B). International Agency for Research on Cancer, Vol. 72. Lyon, France p. 399.

    Google Scholar 

  16. 16.

    IPCS (1985) International Programme on Chemical Safety: Environmental Health Criteria 46. Guidelines for the study of Genetic effects in human populations, WHO, Geneva, 25–54.

    Google Scholar 

  17. 17.

    Joosten, H. F. P., van Acker, F. A. A., van den Dobbelsten, D. J., Horbach, G. J. M. J., Krajnc, E. I. (2004) Genotoxicity of hormonal steroids. Toxicol. Lett. 151, 113–134.

    CAS  PubMed  Google Scholar 

  18. 18.

    Kasper, P., Mueller, L. (1999) Sex specific induction of apoptosis by cyproterone acetate in primary rat hepatocytes. Carcinogenesis 20, 2185–2188.

    CAS  PubMed  Google Scholar 

  19. 19.

    Kasper, P., Tegethoff, K., Mueller, L. (1995) In vitro mutagenicity studies on cyproterone studies on cyproterone acetate using female rat hepatocytes for metabolic activation and as indicator cells. Carcinogenesis 16, 2309–2314.

    CAS  PubMed  Google Scholar 

  20. 20.

    Kedlaya, G. R., Vasedevan, D. M. (2004) Inhibition of lipid peroxidation by botanical extracts of Ocimum sanctum: In vivo and in vitro. Life Sci. 76, 21–28.

    Google Scholar 

  21. 21.

    Keml, M. A., Nair, M. G., Strassburg, G. M., Dewitt, D. L. (2000) Antioxidant and cyclooxygenase inhibitory phenolic compounds from Ocimum sanctum Linn. Phytomedicine 7, 7–13.

    Google Scholar 

  22. 22.

    Lamberti, L., Bigatti-Ponzetto, P., Ardito, G. (1983) Cell kinetics and sister chromatid exchange frequency in human lymphocytes. Mutat. Res. 193–199.

    Google Scholar 

  23. 23.

    Martelli, A., Campart, G. B., Ghia, M., Allavena, A., Mereto, E., Brambilla, G. (1996) Induction of micronuclei and initiation of enzyme altered foci in the liver of female rats treated with cyproterone acetate, chlormadinone acetate or megestrol acetate. Carcinogenesis 17, 551–554.

    CAS  PubMed  Google Scholar 

  24. 24.

    Martelli, A., Mattioli, F., Angiola, M., Reimann, R., Brambilla, G. (2003) Species, sex and interindividual differences in DNA repair induced by nine sex steroids in primary cultures of rat and human hepatocytes. Mutat. Res. 536, 69–78.

    CAS  PubMed  Google Scholar 

  25. 25.

    Martelli, A., Mattioli, F., Fazio, S., Andrae, U., Brambilla, G. (1995) DNA repair synthesis and DNA fragmentation in primary cultures of human and rat hepatocytes exposed to cyproterone acetate. Carcinogenesis 16, 1265–1269.

    CAS  PubMed  Google Scholar 

  26. 26.

    Neumann, I., Thierau, D., Andrae, U., Greim, H., Schwarz, L. R. (1992) Cyproterone acetate induces DNA damage in cultured rat hepatocytes and preferentially stimulates DNA synthesis in γ-glutamyl transpeptidase positive cells. Carcinogenesis 13, 373–378.

    CAS  PubMed  Google Scholar 

  27. 27.

    Perry, P., Wolff, S. (1974) New Giemsa method for differential staining of sister chromatids. Nature 261, 156–158.

    Google Scholar 

  28. 28.

    Reimann, R., Kalweit, S., Lang, R. (1996) Studies for a genotoxic potential of some endogenous and exogenous sex steroid. II. Communication: examination for the induction of cytogenetic damage using the chromosomal aberration assay on human lymphocytes in vitro and the mouse bone marrow micronucleus test in vivo. Environ. Mol. Mutagen. 28, 133–144.

    CAS  PubMed  Google Scholar 

  29. 29.

    Shymala, A. C., Devaki, T. (1996) Studies on peroxidation in rats ingesting copper sulphate and effect of subsequent treatment with Ocimum sanctum. J. Clinical Biochem. Nutr. 20, 113–119.

    Google Scholar 

  30. 30.

    Siddique, Y. H., Afzal, M. (2004) Evaluation of genotoxic potential of synthetic progestin chlormadinone acetate. Toxicol. Lett. 153, 221–225.

    CAS  PubMed  Google Scholar 

  31. 31.

    Siddique, Y. H., Afzal, M. (2004) Evaluation of genotoxic potential of synthetic progestins ethynodiol diacetate in human lymphocytes in vitro. Curr. Sci. 86, 1161–1168.

    CAS  Google Scholar 

  32. 32.

    Siddique, Y. H., Afzal, M. (2005) Genotoxic potential of cyproterone acetate: a possible role of reactive oxygen species. Toxicol. in vitro 19, 63–68.

    CAS  PubMed  Google Scholar 

  33. 33.

    Siddique, Y. H., Afzal, M. (2005) Protective role of allicin and L-ascorbic acid against the genotoxic damage induced by chlormadinone acetate in cultured human lymphocytes. Indian J. Exp. Biol. 43, 769–772.

    CAS  PubMed  Google Scholar 

  34. 34.

    Siddique, Y. H., Beg, T., Afzal, M. (2005) Antigenotoxic effects of ascorbic acid against megestrol acetate induced genotoxicity in mice. Hum. Exp. Toxicol. 24, 121–127.

    CAS  PubMed  Google Scholar 

  35. 35.

    Siddique, Y. H., Beg, T., Afzal, M. (2006) Protective effect of nordihydroguaiaretic acid (NDGA) against norgestrel induced genotoxic damage. Toxicol. in vitro 20, 227–233.

    CAS  PubMed  Google Scholar 

  36. 36.

    Swierenga, S. H. H., Heddle, J. A., Sigal, E. A., Gilman, J. P. W., Brillinger, R. L., Douglas, G. R., Nestmann, E. R. (1991) Recommended protocols on a survey of current practice in genotoxicity testing laboratories, I V. Chromosome aberrations and sister chromatid exchanges in Chinese Hamster Ovary V79, Chinese Hamster lung and human lymphocyte cultures. Mutat. Res. 246, 301–322.

    CAS  PubMed  Google Scholar 

  37. 37.

    Tillmann, M., Schulte-Oehlmann, U., Duft, M., Markert, B., Oehlmann, J. (2001) Effects of endocrine disruptors on prosobranch snails (Mollusca: Gastropoda) in the laboratory part III: Cyproterone acetate and vinclozolin as anti androgens. Ecotoxicology 10, 373–388.

    CAS  PubMed  Google Scholar 

  38. 38.

    Topinka, J., Andrae, U., Schwartz, L. R., Wolff, T. (1993) Cyproterone acetate generates DNA adducts in rat liver and in primary rat hepatocyte cultures. Carcinogenesis 14, 423–427.

    CAS  PubMed  Google Scholar 

  39. 39.

    Topinka, J., Binkova, B., Zhu, H. K., Andrae, U., Neumann, I., Schwartz, L. R., Werner, S., Wolff, T. (1995) DNA damaging activity of cyproterone acetate analogues of chlormadinone acetate and megestrol acetate in rat liver. Carcinogenesis 16, 1483–1487.

    CAS  PubMed  Google Scholar 

  40. 40.

    Torri, V., Floriani, I. (2005) Cyproterone acetate in the therapy of prostatic carcinoma. Arch. Ital. Urol. Androl. 77, 157–163.

    PubMed  Google Scholar 

  41. 41.

    Tucker, J. D., Preston, R. J. (1996) Chromosomes aberrations, micronuclei, aneuploidy, sister chromatid exchanges and cancer risk assessment. Mutat. Res. 365, 147–159.

    PubMed  Google Scholar 

  42. 42.

    Vrinda, B., Devi, P. U. (2001) Radiation protection of human lymphocyte chromosomes in vitro by orientin and vicenin. Mutat. Res. 498, 37–46.

    Google Scholar 

  43. 43.

    Wagner, H., Nörr, H., Winterhoff, H. (1994) Plant adaptogens. Phytomedicine 1, 63–76.

    CAS  PubMed  Google Scholar 

  44. 44.

    Werner, S., Kuntz, S., Beckurts, T., Heidecke, C. D., Wolff, T., Schwarz, L. R. (1997) Formation of DNA adducts by cyproterone acetate and some structural analogues in primary cultures of human hepatocytes. Mutat. Res. 395, 179–187.

    CAS  PubMed  Google Scholar 

  45. 45.

    Yamasaki, K., Sawaki, M., Noda, S., Muroi, T., Takakura, S., Mitoma, H., Sakamoto, S., Nakai, M., Yakabe, Y. (2004) Comparison of the Hershberger assay and androgen receptor binding assay of twelve chemicals. Toxicology 195, 177–186.

    CAS  PubMed  Google Scholar 

  46. 46.

    Yanpallewar, S. U., Rai, S., Kumar, M., Acharya, S. B. (2004) Evaluation of antioxidant and neuroprotective effect of Ocimum sanctum transient cerebral ischemia and long term cerebral hypoperfusion. Pharmacol. Biochem. Behav. 79, 155–164.

    CAS  PubMed  Google Scholar 

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Acknowledgements

Thanks are due to the CSIR, New Delhi for awarding the Fellowship No. 9/112(353)/2003-EMR to the author (YHS) and to the Chairman, Department of Zoology, A.M.U., Aligarh, U.P., for laboratory facilities.

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Correspondence to Y. H. Siddique.

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Siddique, Y.H., Ara, G., Beg, T. et al. Anti-Genotoxic Effect of Ocimum sanctum L. Extract against Cyproterone Acetate Induced Genotoxic Damage in Cultured Mammalian Cells. BIOLOGIA FUTURA 58, 397–409 (2007). https://doi.org/10.1556/ABiol.58.2007.4.7

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Keywords

  • Ocimum sanctum
  • cyproterone acetate
  • anti-genotoxicity
  • human lymphocytes
  • aqueous extract