Russian Journal of PhysiologyRussian Journal of PhysiologyРоссийский физиологический журнал им. И.М. Сеченова0869-81392658-655XThe Russian Academy of Sciences65174510.31857/S0869813924090062AJZMRGEXPERIMENTAL ARTICLESЭКСПЕРИМЕНТАЛЬНЫЕ СТАТЬИResearch ArticleExpression of apoptosis, autophagy and necroptosis effectors in cells of rat hippocampus after excessive F- consumptionЭкспрессия эффекторов апоптоза, аутофагии и некроптоза в клетках гиппокампа крыс после избыточного потребления F-NadeiO. V.НадейО. В.
Russian Federation
nagalak@mail.ruAgalakovaN. I.АгалаковаН. И.
Russian Federation
nagalak@mail.ruSechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of SciencesИнститут эволюционной физиологии и биохимии им. И.М. Сеченова РАН1509202411091362137602022025Copyright ©; 2024, Russian Academy of SciencesCopyright ©; 2024, Российская академия наук2024Russian Academy of SciencesРоссийская академия наукhttps://journals.eco-vector.com/0869-8139/article/view/651745

The work examined the expression of apoptosis, autophagy and necroptosis markers in hippocampal cells of rats after long-term consumption of excessive F- doses at the transcriptional and translational levels. Male Wistar rats were divided into 4 groups receiving 0.4 (control), 5, 20 and 50 mg/l F- (as NaF) for 12 months. The changes in contents of effectors of mitochondrial (Bcl-2, Bax, Caspase-9, Caspase-3) and receptor (Caspase-8, Fas) pathways of apoptosis, mediators (Ulk-1, Beclin-1) and modulators (AMPK, Ark, mTOR) of autophagy, as well as that of necroptosis (RIP and MLKL) were assessed by immunoblotting, the gene expression (Bcl2, Bax, Casp3, Ulk1, Beclin1, Prkaa1, Akt, and mTor) – by real-time PCR. In the hippocampus of F – exposed animals, the expression ratio of Bcl2/Bax genes and Bcl-2/Bax proteins decreased, caspase-9 and caspase-3 were activated, but the level of caspase-8 and membrane Fas receptor remained stable. Long-term F- consumption had no effect on the content of autophagy initiator Ulk-1 and protein kinases AMPK, Akt and mTOR, but resulted in inhibition of key autophagy mediator Beclin-1. The expression level of necroptosis RIP and MLKL effectors in the hippocampal cells of rats received excessive F- did not change as well. Thus, long-term F- exposure was accompanied by activation of apoptosis, mainly through the mitochondrial pathway, at the background of autophagy suppression.

В работе исследовали экспрессию маркеров апоптоза, аутофагии и некроптоза в клетках гиппокампа крыс после длительного потребления избыточных доз F- на уровне транскрипции и трансляции. Самцы крыс Wistar были разделены на 4 группы, получавшие 0.4 (контроль), 5, 20 и 50 мг/л F- (в виде NaF) в течение 12 месяцев. Изменения содержания эффекторов митохондриального (Bcl-2, Bax, каспазы-9, каспазы-3) и рецепторного (каспазы-8, Fas) путей апоптоза, посредников (Ulk-1, Beclin-1) и модуляторов (AMPK, Akt, mTOR) аутофагии, а также некроптоза (RIP и MLKL) в клетках оценивали методом иммуноблоттинга, экспрессию генов (Bcl2, Bax, Casp3, Ulk1, Beclin1, Prkaa1, Akt и mTor) – методом ПЦР в реальном времени. В гиппокампе животных, подвергавшихся действию F-, снижалось соотношение экспрессии генов Bcl2/Bax и белков Bcl-2/Bax, активировались каспаза-9 и каспаза-3, однако уровень каспазы-8 и мембранного рецептора Fas оставался стабильным. Длительное потребление F- не оказало влияния на содержание инициаторного белка аутофагии Ulk-1 и протеинкиназ AMPK, Akt и mTOR, но привело к ингибированию ключевого посредника аутофагии Beclin-1. Уровни экспрессии эффекторов некроптоза RIP и MLKL также не изменялись в клетках гиппокампа крыс, получавших избыток F-. Таким образом, длительное воздействие F- сопровождалось активацией апоптоза, преимущественно по митохондриальному пути, на фоне подавления аутофагии.

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