The Level of Reactive Carbonyl Derivatives of Proteins, Malondialdehyde, and Catalase Activity in the Brain of Rats after Therapy Following Chronic Unpredictable Moderate Stress

BACKGROUND: Understanding the mechanisms of the behavioral disorders’ emergence under the influence of chronic stress is the most important aspect of the subsequent development of a strategy for its therapy and prevention. Changes in the oxidative metabolism processes can be decisive in the development of the pathogenetic cascade in the brain. Information about these processes can be obtained by studying protein carbonylation, lipid peroxidation, and catalase activity (CA). The complexity of the therapeutic impact in various behavioral disorders implies the search for new pharmacological substances and the study of the previously known drugs’ effects based on the available scientific data. AIM: The aim of the study was to study the reactive carbonyl derivatives of proteins (RCDP), malondialdehyde (MDA), and CA in the brain of rats after therapy following chronic unpredictable moderate stress (CUMS). METHODS: Forty male outbred rats weighing 450–500 g were used in this study. For 21 days, all animals were exposed to the diverse stress factors for developing the CUMS. The animals were divided into four groups of 10 rats, each using randomized selection. The rats of one group were euthanized by decapitation with subsequent brain harvesting (Group 4). Remaining three groups of rats were treated with placebo (Group 1), harmine hydrochloride (Group 2), and amitriptyline (Group 3) for 21 days. Upon completion of therapy, all rats were also euthanized by decapitation with subsequent brain harvesting. The levels of RCDP, MDA, and CA were studied in their brain, and after that, we compared the multiple studied indicators in four groups. RESULTS: The results of the rat brain examinations in four groups showed that RCDP level in Group 2 was significantly lower than in Group 4 (p = 0.000). Similarly, in Group 1, it was lower than in Group 4 (p = 0.021), plus, it did not differ statistically from the harmine hydrochloride group (p = 1,000). Indicators of Groups 3 and 4 did not have any significant differences in RCDP level, too, (p = 0.799); however, the RCDP level in Group 2 was significantly lower than in Group 3 (p = 0.040). MDA indicators did not show significant differences; however, a tendency for lower values was revealed in Group 1 (p = 0.233) and Group 2 (p = 0.151). CA in Group 4 was lower than that in Group 1 (p = 0.000), Group 2 (p = 0.001), and Group 3 (p = 0.003) contemporaneously, while all treatment groups were comparable (p = 1.000). CONCLUSION: The result of exposure to chronic stress can be reproduced with the best quality in the CUMS model. The neurobiological foundations of the model make it possible to assess biochemical markers of oxidative metabolism and evaluate the possibilities of pharmacological correction of stress-induced behavioral disorders. To assess the mechanisms of autoregulation of oxidative metabolism, this study included a placebo group (Group 1), the level of RCDP in which was significantly higher in comparison with Group 3 and Group 4 and slightly lower than in Group 2. In this study, harmine hydrochloride demonstrated activity exceeding amitriptyline, particularly limiting the process of protein carbonylation, not noted for amitriptyline. According to the results of the RCDP assessment in the CUMS model, the process of protein carbonylation can be considered to be one of the significant factors in the deactivation of neurotransmitters. The CA levels determined in all groups allowed us to consider this marker as the most sensitive to the effects of stress, which possibly has an inhibitory effect on catalase, as its activity in all groups after therapy was more than two-fold higher than in animals right after CUMS. We can assume that CA plays an important role in starting the processes of autoregulation of oxidative metabolism. The study was carried out as a part of the implementation of the scientific and technical program No. BR05236584 “Development of new herbal preparations and their pharmacological and clinical studies” (O.0820). (2018–2020) in the priority area, “Life and Health Sciences.

Composition of essential oil of leaves and fruits of green strawberry (Fragaria viridis Weston) growing wild in Northern Kazakhstan

Fragaria viridis Weston essential oils from leaves and fruits were obtained by hydrodistillation. The composition of the essential oil from leaves and fruits was analyzed by GC-MS. 39 components were identified in leaves oil representing 67.3-80.7% of the oil composition. The main components of the essential oil from leaves of F. viridis Weston were β-linalool (0.8-8.9%), n-nonanal (0.5-8.6%), tetradecanal (2.1-5.9%), nerolidol (2.1-4.8%), an unidentified component (1.9-6.6%), α-bisabolol (0.8-6.7%), phytol (18.4-47.4%), an unidentified component (0.9-8.2%) depending on the growth conditions. The fruit oil was composed of 34 compounds representing 42.0-70.7% of the total composition of the oil. The main components of the essential oil from fruits of F. viridis Weston were m/p-xylene (2.4-14.0%), isoledene (4.7-8.5%), methyleugenol (3.3-8.4%), α-cedrene (2.5-3.9%), an unidentified component (3.4-9.1%), α-muurolene (6.8-11.3%), nerolidol (1.1-4.8%), α-cedrol (1.7-8.0%), α-bisabolol (2.3-5.0%), an unidentified component (0-25.6%) depending on the growth conditions. This is the first report of the chemical composition of the essential oils obtained from the leaves and fruits of green strawberry (Fragaria viridis Weston)

Фенольные соединения этанольных извлечений Lemna minor L., Lemna trisulca L. и Lemna polyrrhiza L. Schleid. и их иммуномодулирующая активность

The purpose of the study is to determine the composition of the phenolic compounds of ethanol extracts isolated from three species of duckweed: Lemna minor L., Lemna trisulca L. and Lemna polyrrhiza L. (the synonym is Spirodella polyrrhiza Schleid.) and to study its effect on immune system activation.Materials and methods. The objects of the study are: air-dried grass samples of Lemna minor L., Lemna trisuica L. and Lemna polyrrhiza L. collected during their growing season in 2010–2011 in low-flow and stagnant ponds of Kozhevnikovsky and Tomsk districts of Tomsk region. Isolation of polyphenolic complexes (PFC) was carried out by extraction of air-dried raw material with ethyl alcohol. In qualitative and quantitative analysis of the samples studied the method of reversed-phase high-performance liquid chromatography on an Agilent 1100 Series instrument (USA) was used in isocratic mode. In the experiments, 200 male C57BL / 6 and BALB / C mice aged 8–12 weeks were used to determine immunomodulatory activity. Cell proliferation was assessed by a colorimetric method. The absorption of the obtained solutions was measured with a multi-channel spectrophotometer at the wavelength of 540 nm. The determination of antibody-forming cells in the spleen was performed by local hemolysis. The titer of antibodies in serum was evaluated in the hemagglutination reaction. The local hypersensitivity reaction of immediate type was assessed by the author’s modification.Results. For the first time the study of the qualitative composition and quantitative content of PFC of Lemna minor L. (LM) , Lemna trisulca L. (LT) trisulkas, and Lemna polyrrhiza L. (LP) : (4,7 ± 0,4)%, (3,3 ± 0,3)%, (12,8 ± 0,7)% was carried out. The highest content of phenolic acids (10,76%) and the sum of flavonoids, isoflavonoids and coumarins (14,7%) were found in the LP sample. The content of chlorogenic and 3,5-dihydroxybenzoic acids was 2–9 times higher in LP than in other species of duckweed. In LM and LT with concentrations of 5 μg/ml and LP in the range of 0,5–160 μg/ml did not have a toxic effect on antigen presenting cells. When incubated with LT (20 μg/ml), the proliferation of macrophages was reduced by 1,2 times, and when incubated with LM and LT (80 μg/ml), by 1,2 and 1,8 times, respectively. PFC duckweed (LP) did not have a similar effect. LM and LP had a stimulating effect on the production of nitrites in the concentrations of 5 and 20 μg/ml, increasing it by 1,3–1,6 times. Course introduction of LT and LP led to a significant decrease in the number of antibody-forming cells by 1,5 and 2,3 times and a decrease in the local hypersensitivity reaction by 1,9 and 1,5 times, respectively.Цель исследования. Определение состава фенольных соединений этанольных извлечений, выделенных из трех видов ряски: ряски малой (Lemna minor L.), ряски тройчатой (Lemna trisulca L.) и ряски многокорневой (Lemna polyrrhiza L., синоним Spirodella polyrrhiza L.) Schleid.), и изучение его влияния на активацию системы иммунитета.Материал и методы. Объект исследования: воздушно-сухие образцы травы ряски малой (Lemna minor L.), ряски тройчатой (Lemna trisuica L.) и ряски многокорневой (Lemna polyrrhiza L.), собранные в период их вегетации в 2010–2011 гг. в малопроточных и стоячих водоемах Кожевниковского и Томского районов Томской области. Выделение полифенольных комплексов (ПФК) проводили экстракцией воздушно-сухого сырья спиртом этиловым. При качественном и количественном анализе исследуемых образцов применяли метод обращенно-фазовой высокоэффективной жидкостной хроматографии на приборе Agilent 1100 Series (США) в изократическом режиме. В экспериментах для определения иммуномодулирующей активности использовали 200 самцов мышей линий С57ВL/6 и BALB/C в возрасте 8–12 нед. Пролиферацию клеток оценивали колориметрическим методом. Абсорбцию полученных растворов замеряли при помощи многоканального спектрофотометра при длине волны 540 нм. Определение антителообразующих клеток в селезенке проводили методом локального гемолиза. Титр антител в сыворотке крови оценивали в реакции гемагглютинации. Локальную реакцию гиперчувствительности немедленного типа оценивали по методике в авторской модификации.Результаты. Впервые проведено исследование качественного состава и количественного содержания ПФК ряски малой Lemna minor L. (LM), ряски трисульки Lemna trisulca L. (LT) и ряски многокорневой Lemna polyrrhiza L.(LP): (4,7 ± 0,4)%, (3,3 ± 0,3)%, (12,8 ± 0,7)% соответственно. Наибольшее содержание фенолокислот (10,76%) и суммы флавоноидов, изофлавоноидов и кумаринов (14,7%) обнаружено в образце LP. Содержание хлорогеновой и 3,5-дигидроксибензойной кислот в 2–9 раз больше в LP, чем других видах ряски. LM, LT в концентрации 5 мкг/мл и LP в диапазоне 0,5–160 мкг/мл не оказывают токсического действия на антигенпрезентирующие клетки. При инкубации с LT (20 мкг/мл) пролиферация макрофагов снижается в 1,2 раза, а при инкубации с LM и LT (80 мкг/мл) в 1,2 и 1,8 раза соответственно. ПФК ряски многокорневой (LP) не оказывают подобного эффекта. LM и LP оказывают стимулирующее действие на продукцию нитритов в концентрациях 5 и 20 мкг/мл, усиливая ее в 1,3–1,6 раза. Курсовое введение LT и LP приводит к достоверному снижению числа антителобразующих клеток в 1,5 и 2,3 раза и уменьшению величины локальной реакции гиперчувствительности в 1,9 и 1,5 раза соответственно

(1S*,3R*,5S*,7S*)-4,4,8,8-Tetrachloro-1-isopropyl-5-methyltricyclo[5.1.0.03,5]octane

The title compound, C12H16Cl4, is a derivative of the natural product 1-isopropyl-4-methylcyclohexa-1,4-diene, and represents a diastereomer with two trans-fused cyclopropane rings. Both enantiomers are present in the non-centrosymmetric polar space group Pna21. The central cyclohexane ring is planar within 0.02 (1) Å. The C atoms of dichloromethylene groups deviate from this plane by 1.19 (1) and −1.26 (1) Å, whereas the isopropyl and methyl groups are oriented more equatorially, deviating by 0.71 (1) and −0.87 (1) Å, respectively

ЦИТОТОКСИЧЕСКИЕ И ГИПОЛИПИДЕМИЧЕСКИЕ СВОЙСТВА АРГЛАБИНА НА КУЛЬТУРЕ КЛЕТОК ГЕПАТОМЫ КРЫС

Investigated toxicity and lipid-lowering activity (in vitro) of sesquiterpene γ-lactone arglabin as a potential lipid-lowering drugs.Material and methods. Was studied the effect of Arglabin and gemfibrozil (comparison drug) on the lipid level of the rat hepatoma tissue culture (HTC) with fluorescent method with vital dye Nile Red and staining cells with the dye Oil Red O. The cell viability was assessed by using MTT-test and staining with trypan blue.Results. Cultivation of HTC cell cultures with arglabin and gemfibrozil in concentrations up to 50 μm and 0.5 mm, respectively, resulted in dose-dependent decrease in the fluorescence intensity of Nile Red, which is associated with a reduction in lipid level in the cells. In these concentrations the drugs did not exert cytotoxic effect, and HTC viability did not change compared with the corresponding rate in the control culture.Conclusion. Arglabin and gemfibrozil does not affect the viability of hepatoma cells in rats at a concentration up to 50 μm and 0.5 mm, respectively. Arglabin and gemfibrozil dose-dependently reduce the content of lipids in the HTC culture, as evidenced by the decrease in Nile Red fluorescence and decrease lipid droplets stained with Oil Red O in the cytosol.Цель исследования. Оценка in vitro токсичности и гиполипидемической активности сесквитерпенового γ-лактона арглабина в качестве потенциального гиполипидемического средства.Материал и методы. В работе изучали влияние арглабина и гемфиброзила (препарат сравнения) на содержание липидов в клеточной культуре крысиной гепатомы (HTC) флуоресцентным методом с витальным красителем Nile Red и окрашиванием клеток с красителем Oil Red O. Жизнеспособность клеток оценивали с помощью МТТ-теста и окрашиванием с трипановым синим.Результаты. Культивирование клеточной культуры HTC с арглабином и гемфиброзилом в концентрациях до 50 мкМ и 0,5 мМ соответственно приводило к дозозависимому уменьшению интенсивности флуоресценции Nile Red, что связано со снижением содержания липидов в клетках. В этих концентрациях препараты не оказывали цитотоксического действия, и жизнеспособность клеток НТС не изменялась по сравнению с соответствующим показателем контрольной культуры.Заключение. Арглабин и гемфиброзил не влияют на жизнеспособность клеток гепатомы крыс НТС при концентрации до 50 мкМ и до 0,5 мМ соответственно. Арглабин и гемфиброзил дозозависимо снижают содержание липидов в культуре НТС, что подтверждается уменьшением флуоресценции Nile Red и снижением содержания окрашенных Oil Red O липидных капель в цитозоле

Inhibition of T Cell Receptor Activation by Semi-Synthetic Sesquiterpene Lactone Derivatives and Molecular Modeling of Their Interaction with Glutathione and Tyrosine Kinase ZAP-70

A variety of natural compounds have been shown to modulate T cell receptor (TCR) activation, including natural sesquiterpene lactones (SLs). In the present studies, we evaluated the biological activity of 11 novel semi-synthetic SLs to determine their ability to modulate TCR activation. Of these compounds, α -epoxyarglabin, cytisinyl epoxyarglabin, 1 β ,10 α -epoxyargolide, and chloroacetate grosheimin inhibited anti-CD3-induced Ca2+ mobilization and extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation in Jurkat T cells. We also found that the active SLs depleted intracellular glutathione (GSH) in Jurkat T cells, supporting their reactivity towards thiol groups. Because the zeta-chain associated tyrosine kinase 70 kDa (ZAP-70) is essential for TCR signaling and contains a tandem SH2 region that is highly enriched with multiple cysteines, we performed molecular docking of natural SLs and their semi-synthetic derivatives into the ZAP-70 binding site. The docking showed that the distance between the carbon atom of the exocyclic methylene group and the sulfur atom in Cys39 of the ZAP-70 tandem SH2 module was 3.04–5.3 Å for active compounds. Furthermore, the natural SLs and their derivatives could be differentiated by their ability to react with the Cys39 SH-group. We suggest that natural and/or semi-synthetic SLs with an α -methylene- γ -lactone moiety can specifically target GSH and the kinase site of ZAP-70 and inhibit the initial phases of TCR activation

Phenolic compounds of ethanol extracts of Lemna minor L., Lemna trisulca L. and Lemna polyrrhiza L. Schleid and their immunomodulating activity

The purpose of the study is to determine the composition of the phenolic compounds of ethanol extracts isolated from three species of duckweed: Lemna minor L., Lemna trisulca L. and Lemna polyrrhiza L. (the synonym is Spirodella polyrrhiza Schleid.) and to study its effect on immune system activation.Materials and methods. The objects of the study are: air-dried grass samples of Lemna minor L., Lemna trisuica L. and Lemna polyrrhiza L. collected during their growing season in 2010–2011 in low-flow and stagnant ponds of Kozhevnikovsky and Tomsk districts of Tomsk region. Isolation of polyphenolic complexes (PFC) was carried out by extraction of air-dried raw material with ethyl alcohol. In qualitative and quantitative analysis of the samples studied the method of reversed-phase high-performance liquid chromatography on an Agilent 1100 Series instrument (USA) was used in isocratic mode. In the experiments, 200 male C57BL / 6 and BALB / C mice aged 8–12 weeks were used to determine immunomodulatory activity. Cell proliferation was assessed by a colorimetric method. The absorption of the obtained solutions was measured with a multi-channel spectrophotometer at the wavelength of 540 nm. The determination of antibody-forming cells in the spleen was performed by local hemolysis. The titer of antibodies in serum was evaluated in the hemagglutination reaction. The local hypersensitivity reaction of immediate type was assessed by the author’s modification.Results. For the first time the study of the qualitative composition and quantitative content of PFC of Lemna minor L. (LM) , Lemna trisulca L. (LT) trisulkas, and Lemna polyrrhiza L. (LP) : (4,7 ± 0,4)%, (3,3 ± 0,3)%, (12,8 ± 0,7)% was carried out. The highest content of phenolic acids (10,76%) and the sum of flavonoids, isoflavonoids and coumarins (14,7%) were found in the LP sample. The content of chlorogenic and 3,5-dihydroxybenzoic acids was 2–9 times higher in LP than in other species of duckweed. In LM and LT with concentrations of 5 μg/ml and LP in the range of 0,5–160 μg/ml did not have a toxic effect on antigen presenting cells. When incubated with LT (20 μg/ml), the proliferation of macrophages was reduced by 1,2 times, and when incubated with LM and LT (80 μg/ml), by 1,2 and 1,8 times, respectively. PFC duckweed (LP) did not have a similar effect. LM and LP had a stimulating effect on the production of nitrites in the concentrations of 5 and 20 μg/ml, increasing it by 1,3–1,6 times. Course introduction of LT and LP led to a significant decrease in the number of antibody-forming cells by 1,5 and 2,3 times and a decrease in the local hypersensitivity reaction by 1,9 and 1,5 times, respectively