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Vol. 18, Art. 26 (pp. 382-394)    |    2017       

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Acute and subchronic mutagenic and cytotoxic studies of nitrosodimethylamine on retyculocytes and lymphocytes in white rat blood using micronucleus test

Tomilin N.V., Filko O.A., Khrabrova A.V., Solovyeva N.E., Sizova K.V., Utsal V.A., Krasnov K.A.

Institute of Toxicology of Federal Medico-Biological Agency

Brief summary

The objects of this study were lymphocytes and reticulocytes of white rat blood. The aim of this work is to compare the mutagenic and cytotoxic effects of N-nitrosodimethylamine (NDMA) using two kinds of the micronucleus test: on reticulocytes and on lymphocytes using the cytokinesis-block micronucleus (CBMN) assay. The mutagenicity and cytotoxicity were explored after the acute intraperitoneal administration of NDMA in doses of 7.5, 15 and 30 mg/kg at 3 and 24 hours exposure and after daily administration of NDMA in doses of 3.5 and 7 mg/kg for 7 and 14 days. In studies where a micronucleus test on reticulocytes after acute and subchronic administration of NDMA was used, mutagenicity was not detected. The cytotoxic effect on reticulocytes was shown after administering NDMA daily for 7 and 14 days. The decline of the relative content of reticulocytes after the subchronic administration suggests the masking of the mutagenic effect of NDMA by its cytotoxicity. In case of CBMN assay, the mutagenicity of NDMA was detected in 24 hours after administration in doses of 15 and 30 mg/kg, and the cytotoxic effect was detected after an injection of 30 mg/kg. In experiments after subchronic administration of NDMA, mutagenic and cytotoxic effects were revealed after daily injections for 7 days at a dose of 7 mg/kg, and after 14 days the mutagenicity acquired a dose-dependent character after administration in doses of 3.5 and 7 mg/kg. The cytotoxic effect was shown only at a dose of 7 mg/kg. Thus, the obtained results proved the possibility of using lymphocytes in the monitoring studies of the mutagenic action of NDMA. Also proven were the high sensitivity and reliability of the CBMN assay method and its independence from the cytotoxic effect of testing the mutagenicity of NDMA using the micronucleus test on lymphocytes in a cytokinesis-block culture.


Key words

Nitrosodimethylamine, mutagenicity, cytotoxicity, lymphocytes, reticulocytes, micronucleus test.

Reference list

1. N-Nitrosodimethylamine in drinking-water // Background document for development of WHO guidelines for drinking-water quality. - Geneva, Switzerland, 2008. - 29 p.


2. Hakura A., Suzuki S., Sawada S., Sugihara T., Hori Y.J., Uchida K., Kerns W.D., Sagami F., Motooka S., Satoh T. Use of human liver S9 in the Ames test: assay of three procarcinogens using human S9 derived from multiple donors // Regulatory toxicology and pharmacology - 2003. -Vol. 37 - P. 20-27.


3. Baranek D. T. Distribution of methyl and ethyl adducts following alkylation with monofunctional alkylating agents // Mutat. Res. - 1990. - Vol. 231 (1). - P. 11-30.


4. Bowen D. E., Whitwell J. H., Lillford L., Henderson D., Kidd D., Mc Garry S., Pearce?G., Beevers C., Kirkland D. J. Evaluation of a multi-endpoint assay in rats, combining the bone-marrow micronucleus test, the Comet assay and the flow-cytometric peripheral blood micronucleus test. Mutat. Res. - 2011. - Vol. 722. - P. 1-19.


5. Fenech M. The in vitro micronucleus technique // Mutat. Res. - 2000. - Vol. 455 (1-2).?- P. 81-95.


6. Kligerman A. D., Erexson G. L., Wilmer J. L. Development of rodent peripheral blood lymphocyte culture systems to detect cytogenetic damage in vivo // Basic Life Sci. - 1984. - Vol. 29 Pt. B. - P. 569-584.


7. Hayashi M., Morita T., Kodama Y., Sofuni T., Ishidate M., Jr. The micronucleus assay with mouse peripheral blood reticulocytes using acridine orange-coated slides // Mutat. Res. - 1990. - Vol. 245 (4). - P. 245-249.