Assessment of the effect of cell transplantation on DNA repair in rat hepatocytes exposed to carbon tetrachloride (CCl4) by DNA comet assay

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Abstract

Introduction. The effect of carbon tetrachloride (freon-10, asordin, hladon-10) is an organochlorine compound with the chemical formula CCl4 and the subsequent transplantation of fetal liver cells (FLC) on DNA degradation and repair in rat hepatocytes by means of alkaline single-cell gel electrophoresis (DNA comet assay) was assessed.

Material and methods. Acute toxic damage to the rat liver was simulated by a single oral administration to female Wistar rats of CCl4 in an oil solution at a dose of 3000 mg/kg. As a protective agent, a suspension of FLC of E19 rat fetuses was used. Quantitative assessment of the degree of damage to the nuclear DNA of liver cells was performed by DNA comet assay on days 1, 3, 5, 7 and 16 of the experiment.

Results. Intravenous injections of fetal liver cells 6 h after exposure to CCl4 induces DNA repair processes in rat hepatocytes in 5–7 days and led to a decrease in the intensity of nuclear DNA damage. The trend toward a decrease in the number of undamaged hepatocytes continued on the 16th day of the experiment, and, therewith, the enhancement of reparative processes after FLC injection revealed itself in in a significant decrease in the number of hepatocytes with a high intensity of nuclear DNA damage. 

Limitations. To prevent unwanted death of animals in the group, studies were limited to a dose of 3000 mg/kg of CCl4 in oil solution.

Conclusion. The method of alkaline single-cell gel electrophoresis (DNA comet assay) allowed quantitative assessment of the degrees of genome damage and repair. The obtained positive results suggest that FLC exert a protective effect of the structure of the DNA of rat liver role after acute exposure to CCl4. 

Compliance with ethical standards. The study was performed in compliance with the rules of bioethics approved by the European Convention for the Protection of Vertebrate Animals Used for Experimental and Other Purposes. 

Author contribution. All co-authors made an equal contribution to the research and preparation of the article for publication.

Conflict of interests. The authors declare no conflict of interest. 

Acknowledgements. The study was carried out as part of the implementation of the state program of the Russian Federation “Development of healthcare”.

Received: august 9, 2022 / Accepted: January 2023 / Published: February 28, 2023

About the authors

Liliia V. Shabasheva

Research Institute of Hygiene, Occupational Pathology and Human Ecology FMBA of Russia

Author for correspondence.
Email: shabash69@gmail.com

Senior Researcher, Laboratory of General Toxicology and Hygienic Regulation, Research Institute of Hygiene, Occupational Pathology and Human Ecology FMBA of Russia, 188663, Leningrad Region.

e-mail: shabash69@gmail.com

 

Russian Federation

Galina Arkadevna Protasova

Research Institute of Hygiene, Occupational Pathology and Human Ecology FMBA of Russia

Email: galasr@mail.ru
ORCID iD: 0000-0002-5754-3204

Кандидат медицинских наук, ведущий научный сотрудник, ФГУП НИИ гигиены профпатологии и экологии человека ФМБА России, 188663, Санкт-Петербург, Российская Федерация.

e-mail: GalaSR@mail.ru

Russian Federation

Vadim Borisovich Popov

Research Institute of Hygiene, Occupational Pathology and Human Ecology FMBA of Russia

Email: vpopovlr@mail.ru
ORCID iD: 0000-0001-8239-0345

Доктор биологических наук, ведущий научный сотрудник, ФГУП НИИ гигиены профпатологии и экологии человека ФМБА России, 188663, Санкт-Петербург, Российская Федерация.

e-mail: vpopovlr@mail.ru

Russian Federation

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