pharmjournalРазработка и регистрация лекарственных средствDrug development & registration2305-20662658-5049LLC «CPHA»10.33380/2305-2066-2025-14-1-1994pharmjournal-2011Research ArticleДОКЛИНИЧЕСКИЕ И КЛИНИЧЕСКИЕ ИССЛЕДОВАНИЯPRECLINICAL AND CLINICAL STUDIESЭкспериментальный неалкогольный стеатогепатит у мышей ассоциирован с поведенческими, когнитивными и периферическими нейрональными нарушениямиExperimental murine non-alcoholic steatohepatitis is associated with behavioural, cognitive, and peripheral neuronal dysfunctionhttps://orcid.org/0000-0002-4690-1811ПриходькоВ. А.PrikhodkoV. A.

197022, г. Санкт-Петербург, ул. Профессора Попова, д. 14, литера А

14A, Prof. Popova str., Saint-Petersburg, 197022

veronika.prihodko@pharminnotech.comhttps://orcid.org/0009-0003-3505-7904ОрляхинаД. А.OrlyakhinaD. A.

197022, г. Санкт-Петербург, ул. Профессора Попова, д. 14, литера А

14A, Prof. Popova str., Saint-Petersburg, 197022

https://orcid.org/0009-0004-6299-7608ПетроваВ. Д.PetrovaV. D.

197022, г. Санкт-Петербург, ул. Профессора Попова, д. 14, литера А

14A, Prof. Popova str., Saint-Petersburg, 197022

https://orcid.org/0000-0002-7972-1286КаревВ. Е.KarevV. E.

197022, г. Санкт-Петербург, ул. Профессора Попова, д. 9

9, Professora Popova str., Saint Petersburg, 197022

https://orcid.org/0000-0001-9273-6864ИвкинД. Ю.IvkinD. Yu.

197022, г. Санкт-Петербург, ул. Профессора Попова, д. 14, литера А

14A, Prof. Popova str., Saint-Petersburg, 197022

https://orcid.org/0000-0002-9216-8673НапалковаС. М.NapalkovaS. M.

197022, г. Санкт-Петербург, ул. Профессора Попова, д. 14, литера А

14A, Prof. Popova str., Saint-Petersburg, 197022

https://orcid.org/0000-0003-4294-5531ОковитыйС. В.OkovityiS. V.

197022, г. Санкт-Петербург, ул. Профессора Попова, д. 14, литера А

14A, Prof. Popova str., Saint-Petersburg, 197022

Федеральное государственное бюджетное образовательное учреждение высшего образования «Санкт-Петербургский государственный химико-фармацевтический университет» Министерства здравоохранения Российской Федерации (ФГБОУ ВО СПХФУ Минздрава России)Saint-Petersburg State Chemical and Pharmaceutical UniversityФедеральное государственное бюджетное учреждение «Детский научно-клинический центр инфекционных болезней Федерального медико-биологического агентства» (ФГБУ ДНКЦИБ ФМБА)Pediatric Research and Clinical Center for Infectious Diseases of the Federal Medical Biological Agency202526122024141319331Copyright © Приходько В.А., Орляхина Д.А., Петрова В.Д., Карев В.Е., Ивкин Д.Ю., Напалкова С.М., Оковитый С.В., 20252025Приходько В.А., Орляхина Д.А., Петрова В.Д., Карев В.Е., Ивкин Д.Ю., Напалкова С.М., Оковитый С.В.Prikhodko V.A., Orlyakhina D.A., Petrova V.D., Karev V.E., Ivkin D.Y., Napalkova S.M., Okovityi S.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.pharmjournal.ru/jour/article/view/2011Введение

Введение. Неалкогольная жировая болезнь печени (НАЖБП) и неалкогольный стеатогепатит (НАСГ) – на сегодняшний день ведущая причина хронической патологии печени во всем мире – ассоциированы с широким спектром психоневрологических осложнений и заболеваний. Тем не менее в настоящий момент аспекты причинно-следственной связи между патологией печени и нервной системы изучены не до конца, что обусловливает необходимость разработки соответствующих клинически релевантных и валидных животных моделей.

Цель

Цель. Целью настоящей работы стала оценка кратко- и долгосрочных психоневрологических и периферических нейрональных нарушений, осложняющих течение различных стадий НАЖБП/НАСГ у мышей.

Материалы и методы

Материалы и методы. 68 взрослых мышей-самцов линии C57Bl/6 рандомизировали на группы «Контроль» и «НАСГ». НАСГ моделировали в течение 3 (эксперимент 1) или 6 (эксперимент 2) мес. при помощи комбинированной модели, включающей высокожировую диету и введение низких доз тетрахлорметана. Контрольные животные получали стандартный комбикорм, питьевую воду и физиологический раствор в эквивалентных объемах. Поведение животных оценивали через 1, 2, 3 и 6 мес индукции НАСГ в тестах «Открытое поле» (ОП), «Приподнятый крестообразный лабиринт» (ПКЛ) и «Черно-белая камера» (ЧБК). Визуально-пространственную память оценивали с помощью тестов «Спонтанное чередование в Т-лабиринте» и «Распознавание нового объекта» через 1, 2 и 3 мес. индукции НАСГ, а также с помощью теста «Лабиринт Барнс» через 6 мес. индукции НАСГ. Через 3 мес. моделирования НАСГ выполняли игольчатую электронейромиографию (ЭНМГ) икроножной мышцы и двуглавой мышцы плеча при электрической стимуляции седалищного и мышечнокожного нервов соответственно. Патологию печени верифицировали гистоморфологически. Статистическую обработку данных проводили в программе Prism 10.2.3 и среде R 4.2.3 с RStudio 2024.09.0.

Результаты и обсуждение

Результаты и обсуждение. Экспериментальное моделирование сопровождалось снижением общей выживаемости животных (p < 0,05, p < 0,01) и признаками выраженного поражения печени, включая холестатический гепатит, средне- и крупнокапельный стеатоз, очаги некроза и фиброз различной степени тяжести (p < 0,05, p < 0,01). Мыши с НАСГ демонстрировали признаки повышения тревожности в тестах ОП, ПКЛ и ЧБК (p < 0,05, p < 0,01), которые были наиболее выражены на самых начальных (1 мес.) и более поздних (6 мес.) стадиях болезни. НАСГ был также ассоциирован со значительным снижением частоты спонтанного чередования в 3 мес. (p < 0,01), отрицательной дискриминацией объектов в 2 мес. (p < 0,05) и ухудшением ретенции памятного следа в «Лабиринте Барнс» (p < 0,05, p < 0,01) по сравнению с контролем. При проведении ЭНМГ в икроножной мышце у мышей с НАСГ наблюдали значительное снижение максимальной амплитуды М-ответа (p < 0,01) и пороговой силы тока (p < 0,05), в двуглавой мышце плеча – увеличение латентности максимального ответа (p < 0,05).

Заключение

Заключение. Экспериментальный алиментарно-токсический НАСГ у мышей-самцов линии C57Bl/6 ассоциирован с повышением тревожности, нарушением формирования и ретенции визуально-пространственного памятного следа, а также признаками аксонально-демиелинизирующей периферической моторной нейропатии.

Introduction

Introduction. Non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH), the leading causes of chronic liver disease worldwide, are associated with a wide range of psychoneurological complications and conditions. However, the causal relationship between liver and nervous system disease remains poorly understood, which warrants the development of clinically relevant and valid animal models thereof.

Aim

Aim. The objective of this work was to characterize the short- and long-term psychoneurological and peripheral neuronal deficits that complicate different stages of NAFLD/NASH in mice.

Materials and methods

Materials and methods. 68 adult male C57Bl/6 mice were randomized into Control or NASH groups. NASH was induced over 3 (Experiment 1) or 6 (Experiment 2) mo using a combined model including a high-fat diet and low doses of carbon tetrachloride. Control group received standard chow, drinking water, and equivolume normal saline. Animal behaviour was assessed by the Open field (OF), Elevated plus maze (EPM), and Light/dark box (LDB) tests at 1, 2, 3, and 6 mo of NASH induction. Visuospatial memory was assessed by the Spontaneous alternation in the T-maze and Novel object recognition tests at 1, 2, and 3 mo of NASH modelling, and using the Barnes maze at 6 mo of NASH induction. Following 3 mo of NASH induction, needle electroneuromyography (ENMG) was performed on the gastrocnemius and biceps muscles with the electrical stimulation of the sciatic and musculocutaneous nerves, respectively. Liver pathology was confirmed by histomorphology. Statistical analysis was performed using Prism 10.2.3 and R 4.2.3 with RStudio 2024.09.0.

Results and discussion

Results and discussion. Experimental modelling was associated with poorer overall survival (p < 0.05, p < 0.01) and substantial evidence of liver injury, i.e. cholestatic hepatitis, medio- and macrovesicular steatosis, focal necrosis and fibrosis of varying severity (p < 0.05, p < 0.01). Mice with NASH exhibited markers of elevated anxiety in the OF, EPM, and LDB tests (p < 0.05, p < 0.01), which were mostly specific to the very onset of liver disease (1 mo) as well as its later stages (6 mo). NASH was also associated with a significant decrease in spontaneous alternation at 3 mo (p < 0.01), negative object disrimination at 2 mo (p < 0.05), and poorer memory retention in the Barnes maze (p < 0.05, p < 0.01) compared with Control. ENMG data analysis revealed significantly lower peak M-wave amplitudes (p < 0.01) and threshold currents (p < 0.05) in the gastrocnemius, and increased peak latency in the biceps in the NASH group (p < 0.05).

Сonclusion

Сonclusion. Experimental alimentary/toxic NASH in male C57Bl/6 mice is associated with increased anxiety-like behaviour, visuospatial memory acquisition and retention impairment, and evidence of axonal and demyelinating peripheral motor neuropathy.

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