mesЭкстремальная биомедицинаExtreme Medicine3033-89643033-8972Centre for Strategic Planning of the Federal Medical and Biological Agency10.47183/mes.2025-251mes-251Research ArticleКОСМИЧЕСКАЯ МЕДИЦИНАSPACE MEDICINEПризнаки развития окислительного стресса при анализе протеома крови женщин-добровольцев в эксперименте с 5-суточной «сухой» иммерсиейOxidative stress signs in blood proteome analysis of female volunteers in five-day dry immersion testhttps://orcid.org/0000-0002-9646-7275КаширинаД. Н.KashirinaD. N.

Каширина Дарья Николаевна, канд. биол. наук

Москва

Daria N. Kashirina, Cand. Sci. (Biol.)

Moscow

daryakudryavtseva@mail.ruhttps://orcid.org/0000-0002-2071-0443ПастушковаЛ. Х.PastushkovaL. Kh.

Пастушкова Людмила Ханифовна, д-р биол. наук

Москва

Ludmila Kh. Pastushkova, Dr. Sci. (Biol.)

Moscow

lpastushkova@mail.ruhttps://orcid.org/0000-0003-1128-1795БржозовскийА. Г.BrzhozovskiyA. G.

Бржозовский Александр Геннадьевич, канд. биол. наук  

Москва

Alexander G. Brzhozovskiy, Cand. Sci. (Biol.)

Moscow

agb.imbp@gmail.comhttps://orcid.org/0000-0002-2238-3458КононихинА. С.KononikhinA. S.

Кононихин Алексей Сергеевич, канд. физ.-мат. наук

Москва

Alexey S. Kononikhin, Cand. Sci. (Phys. and Math.)

Moscow

konoleha@yandex.ruНиколаевЕ. Н.NikolaevE. N.

Николаев Евгений Николаевич, д-р физ.-мат. наук  

Москва

Evgeniy N. Nikolaev, Dr. Sci. (Phys. and Math.)

Moscow

ennikolaev@rambler.ruhttps://orcid.org/0000-0001-6783-4200ЛаринаИ. М.LarinaI. M.

Ларина Ирина Михайловна, д-р мед. наук

Москва

Irina M. Larina, Dr. Sci. (Med.)

Moscow

irina.larina@gmail.comИнститут медико-биологических проблем РАНРоссияInstitute of Biomedical Problems of the Russian Academy of SciencesRussian FederationИнститут медико-биологических проблем РАН; Сколковский институт науки и технологийРоссияInstitute of Biomedical Problems of the Russian Academy of Sciences; Skolkovo Institute of Science and TechnologyRussian FederationСколковский институт науки и технологийРоссияSkolkovo Institute of Science and TechnologyRussian Federation202526062025272205212Copyright © Каширина Д.Н., Пастушкова Л.Х., Бржозовский А.Г., Кононихин А.С., Николаев Е.Н., Ларина И.М., 20252025Каширина Д.Н., Пастушкова Л.Х., Бржозовский А.Г., Кононихин А.С., Николаев Е.Н., Ларина И.М.Kashirina D.N., Pastushkova L.K., Brzhozovskiy A.G., Kononikhin A.S., Nikolaev E.N., Larina I.M.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/251Введение

Введение. «Сухая» иммерсия, являясь моделью для воспроизведения физиологических эффектов невесомости, позволяет оценить изменения функций сердечно-сосудистой, опорно-двигательной и других систем организма. Исследования протеома крови на основе масс-спектрометрии могут способствовать выявлению механизмов физиологической адаптации к факторам космического полета, моделируемых в «сухой» иммерсии. Цель. Расширение научных представлений о молекулярных участниках острого периода адаптации физиологических систем к условиям моделируемой микрогравитации по данным протеомного анализа сухих пятен крови участниц 5-суточной «сухой» иммерсии.

Материалы и методы

Материалы и методы. В исследовании приняли участие 8 здоровых женщин-добровольцев (средний возраст 30,0 ± 4,8 года). Белки крови анализировали по методу «сухих пятен крови»; использовали капиллярную кровь, высушенную на специальном бумажном фильтре (Perkin Elmer). Ферментное расщепление белков проводили с использованием трипсина (Thermo Scientific, США). Смеси триптических пептидов анализировали методом жидкостной хромато-масс-спектрометрии на хроматографе нано-ВЭЖХ Dionex Ultimate3000, совмещенном с масс-спектрометром TimsTOF Pro. Масс-спектрометрический анализ проводили с использованием метода параллельного накопления при последовательной фрагментации (PASEF). Полученные данные LC-MS/MS были полуколичественно проанализированы с помощью DIA-NN 1.8.1. Статистический анализ проводили в программе Statistica 12.

Результаты

Результаты. Обнаружено, что молекулярный ответ на условия иммерсии связан с повышением уровней белков антиоксидантной защиты, активацией процессов катаболизма и пентозофосфатного пути. Уменьшались уровни отрицательных регуляторов эндопептидаз, белков гомеостаза железа. Выявленные повышенные уровни активаторов НАДФH-оксидазы свидетельствуют об активации НАДФН-оксидазы в условиях эксперимента. Эти результаты могут указывать на развитие окислительного стресса во время иммерсии.

Выводы

Выводы. Выявленные молекулярные участники ответа женского организма на условия иммерсии могут предоставить информацию о сигнальных путях и механизмах, задействованных в ответе на гипокинезию, и в последующем будут способствовать разработке фармакологических мер поддержки здоровья женщин-космонавтов. Эти результаты также могут быть полезны для понимания процессов, приводящих к неблагоприятным последствиям у людей с низким уровнем двигательной активности.

Introduction

Introduction. Dry immersion is a model for reproducing the physiological effects of weightlessness. Such tests allow assessment of changes in the functions of the cardiovascular, musculoskeletal, and other body systems. Mass spectrometry instruments can be used for blood proteome analysis in order to establish mechanisms of physiological adaptation to spaceflight factors simulated in dry immersion tests.

Objective

Objective. To clarify molecular participants in the acute period of adaptation of physiological systems to the conditions of simulated microgravity according to proteome analysis of dry blood spots of participants in a five-day dry immersion test.

Materials and methods

Materials and methods. The study involved eight healthy female volunteers (average age 30 ± 4.8 years). Blood proteins were analyzed using the method of dried blood spots; capillary blood dried on a special paper filter (Perkin Elmer) was used. The enzyme cleavage of proteins was performed using trypsin (Thermo Scientific, USA). Mixtures of tryptic peptides were analyzed by liquid chromatography-mass spectrometry (LC–MS) on a Dionex Ultimate 3000 nano-HPLC combined with a TimsTOF Pro mass spectrometer. Mass spectrometric analysis was performed using the method of parallel accumulation withsequential fragmentation (PASEF). The obtained LC–MS/MS data were semi-quantitatively analyzed using DIA-NN 1.8.1. Statistical analysis was performed in the Statistica 12 software package.

Results

Results. The molecular response to immersion conditions was found to lead to increased levels of antioxidant defense proteins, activation of catabolism processes and the pentose phosphate pathway. The levels of negative regulators of endopeptidases and iron homeostasis proteins decreased. The revealed elevated levels of NADPH oxidase activators indicate activation of NADPH oxidase under experimental conditions. These results may indicate the development of oxidative stress during immersion.

Conclusions

Conclusions. The identified molecular participants in the female body’s response to immersion conditions can provide information about the signaling pathways and mechanisms involved in the response to hypokinesia, and in the future will contribute to the development of pharmacological measures to support the health of female astronauts. These results may also be useful for understanding the processes leading to adverse effects in people with low levels of physical activity.

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