Evaluation of Sprague Dawley rats as biomodels to detect damage on DNA in leukocytes of peripheral blood and hepatic cells, by means of the comet assay.

Authors

  • DF Arencibia Arrebola Instituto Finlay, Vicepresidencia de Investigaciones, Calle 17 e/ 198 y 200, Atabey, Municipio Playa, Apartado Postal 16017, Ciudad de La Habana, Cuba.
  • LA Rosario Fernández Instituto de Farmacia y Alimentos (IFAL, U.H), Calle 222 e/ 25 y 27, Reparto La Coronela, Municipio La Lisa, Ciudad de La Habana, Cuba

Keywords:

Spontaneous, induced, comet assay, strand breaks, alkali-labile sites formation on DNA, Sprague Dawley rats, cyclophosphamide

Abstract

The induction of strand breaks (SB) or alkali-labile sites formation on DNA have been parametersbroadly used for the genotoxicity detection and their implications have been demonstrated indegenerative illnesses, the cancer, and recently linked to the oxidative stress. This article had asobjective to carry out a comparison to the spontaneous basal frequency and induced withcyclophosphamide, the induction of strand breaks or alkali-labile sites formation on DNA ofleukocytes of peripheral blood and hepatic cells of Sprague Dawley rats of both sexes, by means of theComet Assay. We administered 10 rats/sex/group for 14 days, using a negative control group (notadministered), two substance-vehicle controls and positive control cyclophosphamide administeredwith 50 mg/kg, intraperitoneal route. The last time of administered was performed the alkalineelectrophoresis them gel individual cells from leukocytes of peripheral blood and hepatic cells todemonstrate the possible damage to the DNA. These rat line constitute a good in vivo experimental model to demonstrate the primary damage in the DNA given the low spontaneous frequency of theanalyzed indicators. Also under our experimental conditions it is more feasible the biomodels utility inthe determinations of damage in leukocytes of peripheral blood in both sexes.

Downloads

Download data is not yet available.

References

Berwick M, Vineis P. Markers of DNA Repair and Suscentibility to Cancer in Humans: an Epidemiologic Review. J Natl Cancer Inst 2000; 92(11):874-897.

Collins AR. The Comet Assay for ADN Damage and Repair. Principles. Mol. Biotech 2004; 26:249-261.

Lee R, Steinert S. Use of the single gel electrophoresis/comet assay for detecting DNA damage in acuatic (marine and freshwater) animals. Mutat. Res 2003; 544:43-64.

Tice RR, Agurell E, Anderson D, Burlinson B. Single cell gel/Comet assay: Guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 2000; 35:206-221.

Nadin S, Vargas-Roig LM, Ciocca DR. A Silver Staining Method for Single-cell Gell Assay. J. Histochem. Cytochem. 2001; 49(9):1183-1186.

Duez P, Dehon G, Kumps A, Dubois J. Statistics of the Comet assay: a key to discriminate between genotoxic effects. Mutagenesis 2003; 18(2):159-166.

Frieauff W, Hartmann A, Suter W. Automatic analysis of slides processed in the Comet assay. Mutagenesis 2001; 16(2):133-137.

Hartmann A, Agurell E, Beevers C, Brendler-Schwaab S, Burlinson B. Recommendations for conducting the in vivo alkaline Comet assay. Mutagenesis 2003; 18(1):45-51.

Arruzazabala ML, Mas R, Molina V. Effect of D-004, a lipid extract from the Cuban royal palm fruit on atypical prostate hyperplasia induced by phenylephrine in rats. Drugs in R&D 2006; 7:233-241.

Carbajal D, Molina V, Más R, Arruzazabala ML. Therapeutic effect of D-004, a lipid extract from Roystonea regia fruits, on prostate hyperplasia induced in rats. Drugs Exp Clin Res 2005; 31:193-198.

Hipler U, Gorning M, Hipler B, Romer W. Stimulation and scabestrogedn-inducent inhibition of reactive oxygen species generated by rat sertoli cells. Arch Androl 2000; 44:147-154.

Shayne CG. Animal Models in toxicology. In: Published by Shayne C. Gad and Taylor & Francis Group. Toxicology: Chapter 2. The Mouse. 2nd ed. New York: (U.S.A), LLC edition, 2007; p.24-72.

Mitchell AD. Product Safety Evaluation Handbook. In: Marcel Dekker, Inc edition Gad S.C. Genetic Toxicology Testing. U.S.A, 1999; p.167-168.

Wyrobek AJ, Bruce WR. Chemical Mutagens. In: United Kingdom edition published. Principles and Methods for their detection. 2nd ed (vol 2).England, 1978; p.135-136.

González JE, Gámez R, Rodeiro I, García H. Evaluación del efecto genotóxico del D-003 en ratas Sprague Dawley empleando la electroforesis alcalina de células individuales en gel

(Ensayo Cometa). Revista CENIC 2004; 35(2):125-127.

Marrero G, Gutiérrez A, Pardo B, Gámez R, Curveco D. Evaluación del efecto genotóxico del D-004 en ratones NMRI empleando la electroforesis alcalina de células individuales en gel (Ensayo cometa). Revista CENIC 2007; 38(3):200-203.

Arencibia DF, Rosario LA. Algunas consideraciones sobre el desarrollo de la técnica para el ensayo cometa in vivo en leucocitos de sangre periférica y células del hígado. retel (revista de

toxicología en línea) 2010; 26(1):1-12.

Lopes L, Albano F, Laranja GA. Toxicological evaluation by in vitro and in vivo assays of an aqueous extract prepared from Echinodorus macrophyllus leaves. Toxicol Lett 2000; 116:189-

García O, Mandina T. DNA damage evaluated by the comet assay in lymphocytes of children with 137Cs internal contamination caused by the Chernobyl accident. Mut Res 2005; 565(2):191-197.

Rodríguez G, Cancino L, Prieto EA, Espinosa J. El Tinidazol induce roturas de simple cadena en leucocitos de ratón. Anu Toxicol 2001; 1(1):57-64.

Ueno S, Kashimoto T, Susa N, Natsume H, Toya M, Ito N. Assessment of DNA damage in multiple organs of mice after whole body X-irradiation using the comet assay. Mut. Res. 2007; 634(1-2):135-145.

Mani S, Buzaid AC, Cadman EC. Pharmacology of antineoplastic agents, multidrug resistance, and the feature. En: Hoffman R, et al eds. Hematology: basic principles and practice. 2 ed. New York: Churchill Livingstone; 1995.p.915-940.

Alvarez C, Rodríguez R, Wong P, Arévalo J, Canga M, Torres R, Chang L. Nueva Alternativa en el Manejo del Craneofaringioma Sólido Quístico: Bleomicina Intracavitaria más Resección Neuroendoscópica. Anales de la Facultad de Medicina 1999; 60(4):293-297.

Cancino L, Leiva A, Garrido G, Cossío M, Prieto E. VIMANG: los efectos antigenotóxico y modulador de las enzimas glutatión peroxidasa y glutatión-S-transferasa. Rev Cubana Invest

Biomed 2001; 20(1):48-53.

Hartman A, Herkommer K, Glück M, Speit G. DNA-Damaging effects of cyclophosphamide on human blood cells in vivo and in vitro studied with the single-cell gel test (Comet Assay). Environ Mol Mutagen 1995; 25:180-187.

Undeger L, Basaran N, Kars A, Guc D. Assesment of DNA damage in nurses handling antineoplastic drugs by the alkaline COMET assay. Mutat. Res 1999; 439(2):277-285.

Carmignani SS, Raymund GG. Safe handling of cytotoxic drugs in the physician’s office: a procedure manual model. Oncol Nurs Forum 1997; 24 (suppl 1):41-48.

Prieto G, Errecalde C, Trotti N. Farmacología clínica de los antineoplásicos. Monog Med Vet 1999; 19(2):1-8.

Downloads

Published

2010-03-20

How to Cite

1.
Arencibia Arrebola D, Rosario Fernández L. Evaluation of Sprague Dawley rats as biomodels to detect damage on DNA in leukocytes of peripheral blood and hepatic cells, by means of the comet assay. Ars Pharm [Internet]. 2010 Mar. 20 [cited 2024 Jul. 22];51(1):49-56. Available from: https://revistaseug.ugr.es/index.php/ars/article/view/4845

Issue

Vol. 51 No. 1 (2010)

Section

Original Articles

License

The articles, which are published in this journal, are subject to the following terms in relation to the rights of patrimonial or exploitation:

  1. The authors will keep their copyright and guarantee to the journal the right of first publication of their work, which will be distributed with a Creative Commons BY-NC-SA 4.0 license that allows third parties to reuse the work whenever its author, quote the original source and do not make commercial use of it.

b. The authors may adopt other non-exclusive licensing agreements for the distribution of the published version of the work (e.g., deposit it in an institutional telematic file or publish it in a monographic volume) provided that the original source of its publication is indicated.

c. Authors are allowed and advised to disseminate their work through the Internet (e.g. in institutional repositories or on their website) before and during the submission process, which can produce interesting exchanges and increase citations of the published work. (See The effect of open access).