mesЭкстремальная биомедицинаExtreme Medicine3033-89643033-8972Centre for Strategic Planning of the Federal Medical and Biological Agency10.47183/mes.2021.030mes-148Research ArticleОРИГИНАЛЬНОЕ ИССЛЕДОВАНИЕORIGINAL RESEARCHОценка гепатопротекторного эффекта антиоксидантов на амиодарон-индуцированную цитотоксичность в клетках гепатомы человека линии HepaRGAssessing hepatoprotective effects of antioxidants on amiodarone-induced cytotoxicity in human hepatoma HepaRG cell lineФилимоноваК. С.FilimonovaK. S.

ст. Капитолово, к. 93, п. Кузьмоловский, Ленинградская область, 188663

Kapitolovo, 93, p/o Kuzmolovsky, Leningradskaja oblast, 188663

РоговскаяН. Ю.RogovskayaN. Yu.

ст. Капитолово, к. 93, п. Кузьмоловский, Ленинградская область, 188663

Kapitolovo, 93, p/o Kuzmolovsky, Leningradskaja oblast, 188663

БельтюковП. П.BeltyukovP. P.

ст. Капитолово, к. 93, п. Кузьмоловский, Ленинградская область, 188663

Kapitolovo, 93, p/o Kuzmolovsky, Leningradskaja oblast, 188663

БабаковВ. Н.BabakovV. N.

Владимир Николаевич Бабаков

ст. Капитолово, к. 93, п. Кузьмоловский, Ленинградская область, 188663

Vladimir N. Babakov

Kapitolovo, 93, p/o Kuzmolovsky, Leningradskaja oblast, 188663

babakov@gpech.ruНаучно-исследовательский институт гигиены, профпатологии и экологии человека Федерального медико-биологического агентстваРоссияResearch Institute of Hygiene, Occupational Pathology and Human Ecology of the Federal Medical Biological AgencyRussian Federation2021261020242334654Copyright © Филимонова К.С., Роговская Н.Ю., Бельтюков П.П., Бабаков В.Н., 20242024Филимонова К.С., Роговская Н.Ю., Бельтюков П.П., Бабаков В.Н.Filimonova K.S., 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.extrememedicine.ru/jour/article/view/148

Для эффективной терапии амиодарон-индуцированной гепатотоксичности необходимы изучение механизмов токсического действия амиодарона на гепатоциты и оценка возможного влияния гепатопротекторов. Целью работы было исследовать гепатопротекторный эффект антиоксидантов на амиодарон-индуцированную цитототоксичность с использованием иммортализованной гепатомы человека линии HepaRG. Жизнеспособность клеток оценивали при действии амиодарона и в смеси с витамином Е, N-ацетилцистеином и S-аденозилметионином методом импедансометрии, а также определяли содержание некоторых биомаркеров гепатотоксичности с использованием технологии Luminex xMAP. В результате исследования установлен дозозависимый эффект токсического действия амиодарона, IC50 амиодарона для линии HepaRG составил 3,5 мкМ. Показано, что в присутствии витамина Е, N-ацетилцистеина и S-аденозилметионина снижается цитотоксический эффект и увеличивается значение IC50. Амиодарон снижает активность регуляторов клеточного цикла: киназ AKT, JNK и белка p53. В результате действия амиодарона уменьшается содержание АТФ в клетках и наблюдается выход внутриклеточных ферментов (малатдегидрогеназы 1, глутатион-S-трансферазы, сорбитолдегидрогеназы, 5'-нуклеотидазы) в кондиционную среду, что свидетельствует о клеточной гибели по типу некроза. Таким образом, витамин Е, S-аденозилметионин и N-ацетилцистеин снижают цитотоксичность амиодарона в модели амиодарон-индуцированного повреждения гепатоцитов и могут быть рассмотрены в качестве гепатопротекторов при необходимости защиты тканей печени от гепатотоксических эффектов амиодарона.

Effective therapy of amiodarone-induced hepatotoxicity requires studying the mechanisms of the toxic effects of amiodarone on hepatocytes and assessing the potential impact of hepotoprotective agents. The study was aimed to assess hepatoprotective effects of antioxidants on the amiodarone-induced hepatotoxicity with the use of immortalized human hepatoma cells of the HepaRG cell line. Cell viability was evaluated upon exposure to amiodarone and in the mixture with vitamin Е, N-acetylcysteine and S-adenosylmethionine by impedance measurement; the levels of some hepatotoxicity biomarkers were defined using the Luminex xMAP technology. As a result of the research, the dose-dependent toxic effects of amiodarone were established. The IC50 value of amiodarone in the HepaRG cell line was 3.5 μМ. It is shown that cytotoxic effects decrease and the IC50 value increases in the presence of vitamin Е, N-acetylcysteine and S-adenosylmethionine. Amiodarone reduces the activity of cell cycle regulators: AKT, JNK kinases, and p53 protein. Exposure to amiodarone results in reduced intracellular ATP levels and the release of intracellular enzymes (malate dehydrogenase 1, glutathione S-transferase, sorbitol dehydrogenase, 5'-nucleotidase) into conditioned medium, indicating the necrotic cell death. Thus, vitamin Е, S-adenosylmethionine and N-acetylcysteine reduce amiodarone cytotoxicity in the model of amiodarone-induced damage to hepatocytes and can be considered as hepatoprotective agents in case of the need to protect liver against the hepatotoxic effects of amiodarone.

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