toxreviewТоксикологический вестникToxicological Review0869-79223034-4611Federal Scientific Center of Hygiene named after F.F. Erisman10.47470/0869-7922-2022-30-3-158-166toxreview-611Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESРоль цитохромов CYP1A и CYP3A в генотоксическом действии бенз(а)пиренаThe role of cytochromes CYP1A and CYP3A in the genotoxic effect of benzo(a)pyrenehttps://orcid.org/0000-0001-9664-2072МалыгинаДарья АлександровнаMalyginaDaria Alexandrovna

Мл. науч. сотр. лаборатории молекулярной токсикологии и экспериментальной терапии ФГУП «НИИ ГПЭЧ» ФМБА России, 188663, г.п. Кузьмоловский, Ленинградская область, Российская Федерация.

e-mail: malygina.darja@yandex.ru

malygina.darja@yandex.ruРоговскаяНадежда ЮрьевнаRogovskayaNadezhda Yurievna

Науч. сотр. лаборатории молекулярной токсикологии и экспериментальной терапии ФГУП «НИИ ГПЭЧ» ФМБА России, 188663, г.п. Кузьмоловский, Ленинградская область, Российская Федерация.

e-mail: nadin-r@mail.ru

nadin-r@mail.ruhttps://orcid.org/0000-0003-4050-6172БельтюковПетр ПетровичBeltyukovPetr Petrovich

Кандидат мед. наук, зав. лабораторией молекулярной токсикологии и экспериментальной терапии ФГУП «НИИ ГПЭЧ» ФМБА России, 188663, г.п. Кузьмоловский, Ленинградская область, Российская Федерация.

e-mail: beltiukov@gpech.ru

beltiukov@gpech.ruБабаковВладимир НиколаевичBabakovVladimir Nikolaevich

Кандидат биол. наук, ведущий научный сотрудник ФГУП «НИИ ГПЭЧ ФМБА России», 188663, г.п. Кузьмоловский, Ленинградская область, Российская Федерация. 

e-mail: babakov@rihophe.ru

Ph.D., leading researcher, FSUE "Research Institute of Hygiene, Occupational Pathology and Human Ecology", FMBA of Russia, g.p. Kuzmolovskii, 188663, Leningrad Region, Russian Federation. 

e-mail: babakov@rihophe.ru

babakov@rihophe.ruФГУП «НИИ гигиены, профпатологии и экологии человека ФМБА России»РоссияResearch Institute of Hygiene, Occupational Pathology and Human Ecology FMBA of RussiaRussian Federation202220062022303158166Copyright © Малыгина Д.А., Роговская Н.Ю., Бельтюков П.П., Бабаков В.Н., 20222022Малыгина Д.А., Роговская Н.Ю., Бельтюков П.П., Бабаков В.Н.Malygina D.A., Rogovskaya N.Y., Beltyukov P.P., Babakov V.N.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.toxreview.ru/jour/article/view/611Введение

Введение. Метаболиты бенз(а)пирена — генотоксичные соединения, накопление которых способствует канцерогенезу. Основным механизмом их образования является окисление бенз(а)пирена с участием цитохромов P450 (CYP). Ингибиторы основных цитохромов могут приводить к снижению скорости образования метаболитов и, как следствие, к уменьшению генотоксических эффектов метаболитов бенз(а)пирена. Индукторы цитохромов, напротив, способствуют усилению генотоксичности.

Цель исследования — разработка клеточной модели на основе линии HepaRG для исследования роли активности цитохромов в генотоксическом действии бенз(а)пирена.

Материал и методы

Материал и методы. Для оценки влияния ингибиторов цитохромов CYP3A и CYP1A на генотоксические эффекты бенз(а)пирена в клетках HepaRG было определено содержание активных форм белков системы репарации ДНК, фосфорилированных форм белков сигнальных каскадов методом иммуноанализа по технологии Luminex xMAP. Оценку цитотоксичности бенз(а)пирена осуществляли с помощью мониторинга клеточного индекса.

Результаты

Результаты. Ингибиторы цитохромов CYP3A и CYP1A, кетоконазол и α-нафтофлавон снижают токсические эффекты бенз(а)пирена и активацию системы репарации ДНК, оказывают разнонаправленное действие на фосфорилирование различных тирозинкиназ в сигнальных путях.

Заключение

Заключение. Клетки гепатомы человека HepaRG представляют собой подходящую клеточную модель как для оценки вклада цитохромов в метаболизм ксенобиотиков, так и для изучения защиты клеток от генотоксического действия бенз(а)пирена ингибиторами цитохромов.

Ограничения исследования

Ограничения исследования. Исследование выполнено на культуре клеток, для экстраполяции данных на организм требуется учитывать данные токсикодинамики и токсикокинетики.

Introduction

Introduction. Benzo(a)pyrene metabolites are genotoxic compounds, the accumulation of which contributes to carcinogenesis. The main mechanism of metabolite formation is the benzo(a)pyrene oxidation by cytochromes P450 (CYP). Inhibitors of the main cytochromes can reduce the rate of metabolite formation and, as a result, to decrease the genotoxic effects of benzo(a)pyrene metabolites. In contrast, inducers of cytochromes contribute to the enhancement of genotoxicity.

Objective

Objective. The aim of the work was to develop a cell model based on the HepaRG cells to study the role of cytochromes activity in the genotoxic effect of benzo(a)pyrene.

Material and methods

Material and methods. To assess the effect of inhibitors of cytochromes CYP3A and CYP1A on the genotoxic effect of benzo(a)pyrene in HepaRG cells, the content of active forms of proteins of the DNA damage detection and repair system, phosphorylated forms of signaling cascade proteins was determined by immunoassay using Luminex xMAP technology. The cytotoxicity of benzo(a)pyrene was assessed by real-time cell analysis on xCelligence analyzer. 

Results

Results. Inhibitors of CYP3A and CYP1A cytochromes, ketoconazole and α-naphthoflavone demonstrate the ability to diminish the toxic effects of benz(a)pyrene, reduce the activation of the DNA repair system, and have a multidirectional effect on the different tyrosine kinases phosphorylation in signaling pathways.

Conclusion

Conclusion. HepaRG human hepatoma cells are a suitable cell model both to assess the contribution of cytochromes to the metabolism of xenobiotics and to study of the cell protection from the genotoxic effect of benzo (a) pyrene by cytochrome inhibitors.

Limitations

Limitations. The study was performed on a cell culture. To extrapolate the data to the organism, it is necessary to take into account the data of toxicodynamics and toxicokinetics.

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The authors declare that there are no conflicts of interest present.