Elena I. Fedoros

Введение. Разработка новых противоопухолевых препаратов, включая дендритно-клеточные вакцины (ДКВ), требует доклинических исследований (ДКИ), подтверждающих их эффективность in vivo. Это обусловливает необходимость выбора адекватных животных моделей. Цель. Оценить фармакодинамику гомологичного аналога аутологичной вакцины CaTeVac на основе дендритных клеток мыши (ГДКВ) с использованием новой in vivo модели перевиваемой веретеноклеточной саркомы 164 MSar FE (ПВС). Материалы и методы. В предварительном эксперименте сравнивали параметры опухолевого роста и морфологию аутохтонной индуцированной бензо(а)пиреном саркомы (ИБС) и культивируемой ПВС после 11-го, 40-го и 72-го пассажей на мышах линии BALB/c (n = 110, по 55 самцов и самок). В основном эксперименте мыши-самки линии BALB/с получали: 1) ГДКВ в профилактически-лечебном режиме (ПЛ-ГДКВ, n = 13; 1 р/нед. 4 нед. до и 4 нед. после инокуляции ПВС); 2) ГДКВ в лечебном режиме (Л-ГДКВ, n = 13; 1 р/нед. 4 нед после инокуляции); 3) доксорубицин (ДОКС, n = 13, однократно через 48 ч после инокуляции в/б в дозе 8 мг/кг). Контрольная группа (К, n = 12) лечения не получала. Оценивали размеры, ответ опухолей на терапию по модифицированным критериям RECIST 1.1, время до прогрессирования (ВДП) и общую выживаемость (ОВ). Результаты. По результатам предварительного эксперимента был выбран 72-й пассаж ПВС, который характеризовался одномоментным появлением опухолей, коротким латентным периодом (7-9 суток против 83 у ИБС), 100 %-ной перевиваемостью, меньшей в 2,5 раза скоростью роста по сравнению с ИБС. Оптимальных объемов опухоли достигали на 11-14-е сутки против 3 мес. у ИБС, при этом ПВС сохраняла морфологические признаки ИБС. В основном эксперименте профилактически-лечебный режим ГДКВ обеспечивал ответ опухоли на терапию (p = 0,0052 против контроля) у 53,9 % животных, увеличивал медиану ВДП (63 против 32 сут в группе контроля) и ОВ (3/13 животных выжило без прогрессирования). Эффективность лечебного режима ГДКВ и доксорубицина была ниже. Заключение. Охарактеризована и апробирована новая модель ПВС мыши 164 MSar FE (72-й пассаж), на которой показано противоопухолевое действие профилактически-лечебного режима введения ГДКВ.

The aim of this study was to examine the hepatoprotective activity of multicomponent mixtures of natural origin in the BALB/C mouse model (n = 59), with acute direct toxic liver injury (DTLI) induced by the administration of streptozotocin (STZ) (100 mg/kg) in combination with a high-fat and high-fructose diet (HFFD). The hepatoprotective activity of activated hydrolytic lignin (Bp-Cx-1), methanolic fraction of Bp-Cx-1 (Bp-Cx-M) and isoflavones from kudzu Pueraria lobata roots (IFL) were evaluated on molecular level using mass spectrometry (MS)-based omics technologies. Untargeted label-free DIA quantitation resulted in 7214 protein groups identification (FDR 1%) after filtering across 40 liver tissue extracts. All treatment groups were closer to the control samples on the liver proteomic landscape compared to the untreated DTLI group, with the best results shown for the Bp-Cx-M and IFL groups. In order to identify differences between specific groups, we applied the post hoc Dunn’s test and used Hedges’ g as the effect size metric, revealing 64 proteins that tended to return to their normal level after treatment. In-depth proteomic liver tissue analysis enabled us not only to reveal the main pathways such as inflammation and oxidative stress, which are in a good agreement with DTLI and non-alcoholic liver disease pathophysiology, but also to evaluate hepatoprotective activity of multicomponent mixtures of natural origin containing polyphenols and mostly associated with protein metabolism (e.g., PSMD7, HCFC1) and deubiquitination pathways (e.g., UCHL3). It is worth noting that the Bp-Cx-1 isolated methanol fraction (Bp-Cx-M) demonstrated a pronounced increased hepatoprotective activity compared to the parent material due to the enrichment with active components such as polyphenols. Consistent with the proteomic findings of restored ubiquitin–proteasome function, assessment by comet assay revealed that treatments with Bp-Cx-M and IFL significantly reduced DNA damage by 50% compared to the untreated DTLI group. The developed MS-based multi-omics approach may be implemented for the robust and high-throughput screening method during assessment of new hepatoprotective agents of synthetic or natural origin.

BACKGROUND: Experimental models of non-lethal, whole-body gamma irradiation are widely used to study long-term effects of radiation exposure, such as induced carcinogenesis and accelerated aging. The sensitivity of mice to ionizing radiation, which induces lymphoma and reduces lifespan, is known to decrease as the age at the onset of exposure increases. However, the nature and trends in the radiation response of experimental animals remain unclear. AIM: The study aimed to evaluate the factors affecting the lifespan and carcinogenesis parameters of mice subjected to a four-fold total weekly gamma irradiation at a dose of 1.7 Gy per week (total dose: 6.8 Gy). METHODS: The experiments were conducted on 111 male and 70 female CB6F2 mice, with a mean age of 62.7 ± 13.7 days at the start of the study. Body weight, time of death, and carcinogenesis parameters were assessed. An additional group of males (n = 21) was irradiated on days 56–57 of life. Peripheral blood cell composition was analyzed, and a DNA comet assay was performed. RESULTS: Thymic lymphomas developed in 59% of males after 348.4 ± 135.5 days and in 48% of females after 349.5 ± 107.7 days. The mean lifespan was 352.8 ± 137.4 days for males and 349.3 ± 115.0 days for females. Carcinogenesis parameters did not depend on the age at the exposure onset. A positive correlation was found between lifespan and age at first irradiation (r = 0.3020, p = 0.0013 for males, r = 0.3522, p 0.0001 for females), but not with initial body weight. Irradiation of animals at 56–57 days of age at 1.7 Gy resulted in a 1.6-fold increase in the percentage of DNA in the comet tail after the first exposure and a 2.3-fold increase after the fourth, compared to pre-irradiation levels. The granulocyte-to-lymphocyte ratio in peripheral blood increased 1.6-fold after the first exposure and 2.26-fold after the fourth. The values obtained after the first irradiation were inversely correlated with lifespan (r = −0.5201, р = 0.0157). CONCLUSION: In a model of fractionated four-fold gamma-irradiation at a total dose of 6.8 Gy, two critical factors associated with the lifespan of mice are the age at the first session and the resulting peripheral blood granulocyte-to-lymphocyte ratio. In this model, body weight at the time of the first irradiation, the level of DNA damage as assessed by the comet assay, and the development of lymphomas were not associated with the animals' lifespan.

The aim of this study was to examine the hepatoprotective activity of multicomponent mixtures of natural origin in the BALB/C mouse model (59 female mice), with non-alcoholic fatty liver disease (NAFLD) induced by the administration of streptozotocin (STZ) in combination with a high-fat, high-fructose diet. The hepatoprotective activity of activated hydrolytic lignin (BP-Cx-1), metanolic fraction of BP-Cx-1 (BP-Cx-M) and isoflavones from kudzu Pueraria lobata roots (IFL) was evaluated using mass spectrometry (MS)-based omics technologies. Untargeted label-free DIA quantitation resulted in 8088 protein groups identification (FDR 1%) in 40 liver tissue extracts. All treatment NAFLD groups were closer to the control samples on the liver proteomic landscape with the best results shown for BP-Cx-1-m and ISF groups. This corroborated well with metabolomic fingerprinting by FTICR MS. In order to identify differences between specific groups, we applied the post-hoc Dunn’s Test and used Hedges' g as the Effect Size metric and 64 proteins that tended to return to their normal level after treatment were revealed. Pathophysiologically specific proteomics changes occurring during NAFLD were mostly associated with metabolism of the proteins (PSMD7, HCFC1) and deubiquitination pathways (UCHL3). It is worth noting that BP-Cx-1 isolated methanol fraction (Bp-Cx-m) demonstrated the pronounced increased hepatoprotective activity due to the enrichment with active components such as polyphenols. Additionally, it was shown that treatments with Bp-Cx-M and IFL significantly reduced DNA damage by 50% compared to the NAFLD untreated group. Finally, MS-based multiomics approach demonstrated its power as an advanced screening method for the assessment of the hepatoprotective activity and molecular mechanisms of action of complex multicomponent mixtures of natural origin.

Exposure to ionizing radiation during antitumor radiotherapy can cause undesirable side effects. This is damage to healthy tissues and organs. The use of radioprotective drugs can provide protection for normal cells, but there are risks of their negative impact on the antitumor effect of radiotherapy. The aim of the work is to study the possibility of using promising biotechnological radioprotective drugs to reduce the side effects of antitumor radiotherapy.Material and methods. The study was performed on model of Ehrlich’s carcinoma in BALB/c mice with experimental radiation treatment (4 Gy X-ray). Biotechnological products, developed by Russian specialists, were administered to mice before irradiation on the 12th day after the tumor was transplanted. Chitin-glucan complex extracted from Pleurotus ostreatus was administered at a dose of 500 mg/kg intragastrically, recombinant flagellin at a dose of 1 mg/kg intraperitoneally. The volume of tumor nodules was assessed at different time points during the experiment, the degree of tumor growth inhibition was calculated, and the spread of the tumor into nearby tissues was recorded.Results and discussion. The absence of stimulation of tumor growth during the use of drugs has been established. The drugs did not reduce the inhibition of Ehrlich tumor growth in mice caused by experimental X-ray treatment. In some periods of the experiment, mice with combination therapy showed a tendency to increase the effectiveness of radiation treatment by 4–6 %. The use of recombinant flagellin led to a decrease in the number of animals with a total spread of Ehrlich’s carcinoma. When using the mushroom chitin-glucan complex, there was a tendency to decrease the value of the integral dissemination score compared to control.Conclusions. It was shown that the chitin-glucan complex from the Pleurotus ostreatus and the deimmunized recombinant flagellin did not have a negative effect on the antitumor effect of radiation exposure. The possibility of their use as a means of accompanying radiotherapy of tumors has been experimentally confirmed.

Введение. Экспериментальные исследования на млекопитающих указывают на возможный риск сохранения нарушений, вызванных облучением родителей, у потомков не только первого, но и второго поколения. В связи с этим важной задачей является поиск средств для смягчения данного эффекта. Цель. Оценка трансгенерационного влияния полифенольного препарата BP-C2 на формирование опухолей у рыб Danio rerio (зебрафиш) поколения F2, ведущих происхождение от облученных самцов. Материалы и методы. Поколение F2 было получено при скрещивании интактной самки с самцами генерации F1, ведущими свое происхождение от самцов, подвергнутых рентгеновскому облучению в дозе 5 Гр и обработанных или необработанных в растворе препарата BP-C2, а также от контрольных интактных самцов. Рыбы генерации F2 в возрасте 10 мес. были подвергнуты воздействию N-диэтилнитрозоамина (100 ppm) в течение 20 дней. Эвтаназию проводили через 6 мес. после завершения экспозиции с канцерогеном; после гистологической обработки оценивали частоту опухолей печени различного типа. Результаты. У рыб генерации F2, ведущих происхождение от облученных рыб, обнаружено увеличение частоты возникновения гепатоцеллюлярных карцином в 2,75 раза (р = 0,0213), опухолей в целом в 1,86 раз (р = 0,0329), а также злокачественных опухолей в 2,17 раз (р = 0,0128) по сравнению с потомками интактных рыб. Использование BP-C2 после облучения самцов F0 приводило к снижению количества злокачественных опухолей до уровня контроля (р = 0,7300) и к изменению структуры опухолевой патологии: обнаружено уменьшение частоты гепатоцеллюлярных карцином, а также тенденция к увеличению частоты аденом печени и холангиокарцином по сравнению с другими группами. Заключение. Обнаружено, что трансгенерационные последствия радиационного воздействия у рыб Danio rerio сохраняются у потомков облученных самцов во втором поколении. При этом использование полифенольной композиции BP-C2 сразу после облучения рыб уменьшает тяжесть индуцированного диэтилнитрозоамином канцерогенеза печени у потомков.

Введение. Изучение феномена трансгенерационных эффектов ионизирующего излучения насчитывает не один десяток лет. Одним из возможных последствий облучения родителей может быть изменение частоты и спектра развития злокачественных новообразований у потомства, при этом данный вопрос освещается в небольшом числе публикаций экспериментальных и эпидемиологических исследований. Целью данного обзора было определить влияние преконцептивного облучения отцов и матерей на риск развития опухолевой патологии у их потомства. Материалы и методы. В базах данных PubMed и eLIBRARY.RU проведен поиск без ограничений по дате с использованием терминов: «трансгенерационный канцерогенез», «радиационный канцерогенез» в eLIBRARY.RU и transgeneration AND irradiation в PubMed; всего были найдены 236 публикаций. После первоначального ознакомления с названием и аннотацией исключены 161. Для детального анализа отобраны 75 работ, из которых исключены 63 как не соответсвующие критериям, включая одну дублирующую результаты и 22 обзорные публикации. Дополнительный анализ списков литературы в обзорных работах позволил идентифицировать еще 18 публикаций. Всего в систематический обзор включены 30 публикаций. Результаты. В основном, исследования заболеваемости у потомства облученных людей не выявили каких-либо значимых рисков для здоровья. В то же время эксперименты, выполненные на мышах, свидетельствуют в пользу наличия увеличенного риска, как спонтанного, так и индуцированного дополнительным воздействием канцерогенеза у потомства облученных животных. Выводы. В целом совокупность эпидемиологических данных у людей и экспериментальных у мышей не позволяет исключить того, что воздействие облучения на зародышевую линию вызывает слабые прямые онкогенные эффекты, а также сопровождается гиперчувствительностью потомства к последующим канцерогенным воздействиям в постнатальном периоде.

The aim of this study was to examine the hepatoprotective activity of multicomponent mixtures of natural origin in the BALB/C mouse model, with subacute liver failure (SALF) induced by the administration of toxin carbon tetrachloride (CCl₄). The hepatoprotective activity of activated hydrolytic lignin (BP-Cx-1), humic acid peloids (HA), and isoflavones from kudzu Pueraria lobata roots (IFL) was evaluated using mass spectrometry (MS)-based omics technologies. Our MS-based approach revealed new insights into the molecular mechanisms of the hepatoprotective activity of multicomponent mixtures of natural origin. Significant differences were observed in the proteome and metabolome profiles of the urine and liver of BALB/c mice with SALF between a control group with CCl₄ administration, intact controls, and groups receiving potential hepatoprotectors of natural origin (BP-Cx-1, HA, IFL). Proteomic and metabolomics analyses demonstrated that among the hepatoprotectors, IFL possessed the highest hepatoprotective potential, which correlated well with the relative effectiveness of the drugs recorded during in vitro studies. These results correlate with the relative effectiveness of the drugs recorded in previous in vitro and in vivo studies. The leading IFL activity may be attributed to a higher content of active polyphenolic components compared to heterogeneous HA and BP-Cx-1. Enrichment with active components by fractionation is a direction that can be explored for developing hepatoprotective agents based on natural complex polyphenols.

The aim of this study was to examine the hepatoprotective activity of multicomponent mixtures of natural origin in the BALB/C mice model with subacute liver failure (SALF) induced by the administration of toxin carbon tetrachloride (CCl4). The hepatoprotective activity of activated hydrolytic lignin (BP-Cx-1), humic acid peloids (HA) and isoflavones from kudzu, Pueraria lobata (IFL) roots was evaluated using mass spectrometry based omics technologies. MASS SPECTROMETRY BASED multi-OMICS analysis revealed new insight on the molecular mechanism of the hepatoprotective activity of multicomponent mixtures of natural origin. The significant differences were observed in proteome and metabolome profiles of urine and liver of BALB/c mice with SALF between control group with CCl4 administration, intact control, and groups receiving potential hepatoprotectors of natural origin (BP-Cx-1, HA, IFL). Proteomic and metabolomics analysis demonstrated that among samples IFL possessed the highest hepatoprotective potential, which correlated well with the relative effectiveness of the drugs recorded during in vitro studies. The results correlate with the relative effectiveness of the drugs recorded in previous in vitro and in vivo studies. Leading IFL activity may be attributed to a higher content of active polyphenolic components compared to heterogeneous HA and BP-Cx-1. Enrichment with active components by fractionation is a direction, which can be explored for developing hetepoprotective agent based on natural complex polyphenols

A model for accelerated aging in mice was developed: CB6F2 mice aged 39-45 days were exposed to fractionated 4-fold relatively uniform γ-radiation (13