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Vol 23, No 2 (2020)

Articles

Variability of the component composition of Vitexagnus-castus L. essential oil

Shevchuk O.M., Feskov S.A., Logvinenko L.A., Fedotova I.A.

Abstract

Vitex agnus-castus L. (chaste tree) is a valuable medicinal and essential oil plant (Lamiaceae), cultivated as an ornamental and medicinal plant in subtropical regions around the world. Under the natural conditions of the Crimea, chaste tree grows singly or in groups at the mouths of beams on the coast, is listed in Red Book of the Crimea with environmental status - a rare species. In the collections of the Nikita Botanical Gardens there are three forms of Vitex introduced from different places of growth and differing in morphological and biological characteristics. Vitex leaves contain essential oil, characterized by antimicrobial, antifungal, anti-inflammatory effects, as well as high antibacterial activity. The aim of our research was to investigate the component composition of the essential oil of the leaves of these forms of chaste tree to determine their chemotypes. Material and Methods. The mass fraction of essential oil was determined during the budding phase of plants (early July) by hydrodistillation on Ginsberg apparatuses. The composition of the volatiles was determined using a hardware-software complex based on a Chromatek-Crystal 5000.2 chromatograph. Results. As a result of the studies, it was found that the form with a light purple color of the corolla, introduced from the natural flora of the Crimea, in the conditions of culture is characterized as long-growing with a high content of essential oil in the leaves (0.79% of the absolute dry weight) with a sharp pepper-clove spicy odor due to the high proportion of sesquiterpenes (β-caryophylllene, β-farnesene, bicycloher-macrene), the amount of which is 43%. The foreign forms of chaste tree with a pink and purple color of the corolla are distinguished by an early onset of flowering, a lower content of essential oil (0.47-0.6%) with a strong balsamic peppermint smell and a high content of 1,8-cineole in it (35, 2-37.7%). Conclusions. A comparative analysis of the obtained and existing literature data indicates the presence of several chemotypes of essential oil in chaste tree: 1,8-cineole and pinene. The essential oil of the studied forms of chaste tree from the natural habitats of the Crimea belongs to the cineole-farnesene subtype, and foreign introduced ones - to the cineole-sabinene.
Problems of Biological Medical and Pharmaceutical Chemistry. 2020;23(2):3-11
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Collection of cell cultures as an alternative source of biologically active substances

Tsybulko N.S., Myasnikova S.B.

Abstract

VILAR is the holder of a collection of 16 strains of cultures of nine medicinal plants species: Macleaya cordata Willd, Thalictrum minus L., Stephania glabra Roxb., Miers, Podophyllum peltatum L., Panax ginseng C. A.Mey, Rodiola rosea L., Macrotomia eychroma Royle Pauls, Ungernia victoris Vved.ex Artjuschenko, Hedysarum neglectum. The presented cultures contain a wide range of biologically active substances: alkaloids (berberine, sanguinarine, helerubin, stefarin, galantamine); saponins, phenylpropanoids, lignans (podophyllotoxin), glycosides (mangiferin), shikonin. During cultivation, cell strains are maintained in the form of a suspension culture in a nutrient medium according to the Murasige-Skoog prescriptions with the addition of the necessary vitamins and growth regulators and in the form of callus on agar. The viability of the crops is maintained on a hanging rocking chair with a frequency of 100 rpm in a dark room at a temperature of 26° C and in thermostats TC80M2. To overcome the instability of the processes of growth and synthesis of target secondary metabolites during long-term cultivation, cell strains are maintained under strictly controlled conditions in accordance with the cultivation technology - periodic transplants and regular monitoring of the content of the main active substances. Qualitative and quantitative analysis of the target components is carried out by a chromatospectrophotometric method. We find, that thalictrum minus biomass productivity and berberine biosynthesis (up to 0.10-0.12%) are kept a long time (more than 13 years). The maximum synthesis of alkaloids sanguinarine and helerubine (up to 0.88%) in total cell culture of macleaya is kept more than 8 years. This confirms the sufficient stability of the obtained strains. There was also find the variation in the level of maximum accumulation of target metabolites in the process of cultivation during the year with the maximum level at spring -sampler season.
Problems of Biological Medical and Pharmaceutical Chemistry. 2020;23(2):12-17
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Flakozid - drug from the leaves of Phellodendron amurense Rupr., in treatment of diseases of the hepatobiliary system

Bortnikova V.V., Karabaeva V.V., Krepkova L.V., Sidelnikova G.F., Mizina P.G., Karabaeva O.N.

Abstract

The aim: to show the effectiveness of the drug Flakozid, which has a broad spectrum of antiviral action in combination with a hepato-protective effect, in the treatment of diseases of the hepatobiliary system. Material and Methods. The analysis of the effectiveness of the drug has been carried out based on the results of clinical studies of Flakozid, presented by Russian clinical institutions. Results. With chronic hepatitis and cirrhosis of the liver, unconjugated hyperbilirubinemia and fatty liver, the use of oral Flakozid in daily doses of 0.3-0.6 g for 25-45 days has led to an improvement in the general condition of patients, reduction of pain in the right hypochondrium, reduction of dyspeptic disorders, reduce the effects of asthenia, increase physical and intellectual performance, improve appetite. Noted a decrease in the level of total bilirubin and the normalization of the level of ALAT, ASAT and alkaline phosphatase. According to cholecystography, the concentration and contractility of the gallbladder improved. In terms of the severity of the therapeutic effect, Flakozid was not inferior to Silibour and Karsil, and in some indicators (improvement in motility of the gallbladder and biliary tract) markedly exceeded them. Conclusions. According to the results of clinical studies, it has been established that treatment of Flakozid has improved the functional state of the liver, reducing the syndrome of cytolysis and cholestasis. Flakozid has recommended in clinical practice in the complex treatment of diseases of the hepatobiliary system, such as CAG, chronic persistent hepatitis, chronic stoneless cholecystitis at the stage of subcompensation and decompensation.
Problems of Biological Medical and Pharmaceutical Chemistry. 2020;23(2):18-23
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Specific features of serum amino acid concentration in preschool children with autism spectrum disorder

Skalny A.V., Korobeynikova T.V., Skalny A.A., Lobanova Y.N., Skalnaya M.G., Tinkov A.A.

Abstract

The aim of this study was to investigate the serum concentration of amino acids, as well as their derivatives, in children with an autism spectrum disorder of preschool age. Material and methods. There are 35 boys with ASD (ICD-10: F84.0) of preschool age (3-6 years) and 35 healthy peers were examined. Determination of serum concentration of amino acids and their derivatives was carried out by high performance liquid chromatography (HPLC) on a PerkinElmer S200 chromatograph (PerkinElmer, USA). Results. It was found that the concentration of leucine, lysine, threonine, tryptophan, arginine, glutamine, serine, 1-methylhistidine, α-aminobutyric acid and taurine was characterized by a significant decrease relative to the control values by 11%, 22%, 19%, 22%, 34%, 17%, 13%, 29%, 20% and 14%, respectively. It is also worth noting a 16% decrease in serum phenylalanine, approaching significant in significance. At the same time, the hydroxyproline serum concentration in children with autism exceeded the corresponding parameters in the control group by more than 3 times. In accordance with the results of a group comparison, a tight relationship with ASD was found for the concentration of hydroxyproline (β = 0.447; p = 0.001) and arginine (β = -0.238; p = 0.068). A model including the level of amino acids characterized by any significant group differences (1mh, Aab, Arg, Gln, Hypro, Leu, Lys, Ser, Thr, Trp, Tau) was associated with 26% of the variability of the presence of ASD (p = 0.001). Conclusions. Thus, the data obtained indicate a pronounced trend towards amino acid deficiency in children with ASD amid an increase in the concentration of hydroxyproline, which may be the link between autism and joint hypermobility. In turn, a deficiency of a number of amino acids, which are precursors of neurotransmitters, can lead to aggravation of neuropsychiatric disorders. In this regard, the correction of the amino acids metabolism may be one of the potential directions in the treatment of ASD.
Problems of Biological Medical and Pharmaceutical Chemistry. 2020;23(2):24-30
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The influence of pumpkin polysaccharides on oxidative stress indices in acute hypoxia with hypercapnia in rats

Marsyanova Y.A., Laksaeva E.A., Karimov I.A., Zvyagina V.I.

Abstract

The aim. In the model of acute hypoxia with hypercapnia in an experiment on male rats, the protective effect of polysaccharides of pumpkin fruits in relation to the oxidative modification of protein molecules of skeletal muscle tissue is shown. Material and Methods. The experiment examined tissues of the cardiac and skeletal muscles taken from 18 sexually mature male rats of Wistar weighing 200-220 g, which were kept in the vivarium of Ryazan State Medical University. The animals were divided into three groups of 6 individuals. The first group of animals received a 10% solution of polysaccharides extracted from pumpkin fruits by intragastric feeding through a tube daily for 30 days at the dose of 0.1 g / kg body weight. The polysaccharides were extracted according to the following scheme: to remove extractives and colored molecules from the raw material, it was preliminarily treated with 4060% ethanol solutions. Then, a weighed portion of air-dried fruits was placed in a round bottom flask. After that it was poured with a 1% solution of ammonium oxalate and extracted in a boiling water bath for one and a half hours. After that the mixture was filtered, the filtrate was evaporated, and the obtained extract was precipitated with one and a half volume of 96% ethanol. The precipitate (PS) was separated, purified with ethanol, ether and acetone. The isolated PS was dried in vacuo for 12 hours over P2O5. On the last day of PS administration, the animals were exposed to acute normobaric hypoxia with hypercapnia once. For this, each animal was placed in a sealed chamber with a volume of 1000 ml for 30 minutes. The second group of animals received saline according to the introduction of PS. On the last day of the experiment, the animals were placed in a chamber with natural ventilation for 30 minutes. The third group of the animals remained intact until the end of the experiment and underwent hypoxia in the same way as the first group of the animals. After that tissues were taken from the anesthetized rats. Homogenates were obtained from skeletal and cardiac muscle tissues using a homogenizer, which were centrifuged for 10 minutes at 1000 g to precipitate nuclei and intact cells. The resulting supernatant was used for further studies. To assess the antioxidant ability of PS, the activity of the enzyme of the antioxidant system of cells, superoxide dismutase (SOD), was determined; to evaluate the protective effect of PS in acute hypoxia with hypercapnia, oxidative modification of proteins (OMB) was determined as markers of oxidative stress. For calculations in each sample, the concentration of total protein was determined by the Lowry method. Statistical data processing was performed using Microsoft Office Excel 2010. The level of differences was considered statistically significant with an error probability of p <0.05. Results. The results of evaluating the activity of SOD show that the state of hypoxia with hypercapnia, modeled in the experiment, causes an increase in enzyme activity by 2.74 times in the heart muscle and 2.34 times in the skeletal muscle. At the same time, the activity of SOD during hypoxia while taking polysaccharides is reduced by 1.52 times in the heart muscle and by 2.55 times in the skeletal muscle. Determination of the oxidative modification of proteins showed that polysaccharide intake did not influence the modification of the heart muscle proteins. However, the development of hypoxia in this model did not lead to the accumulation of oxidized modified derivative proteins. In skeletal muscle, the total area under the curve of the absorption spectrum of the oxidized modified protein during the development of hypoxia turned out to be 4.26 times greater, with the greatest increase in derivatives of ketone-dinitrophenolhydrozone (KDNPH). However, with PS administration, the accumulation of modified proteins was 2.05 times less than with hypoxia. PS administration reduced the accumulation of aldehyde-dinitrophenolhydrozone (ADNPH) of a basic and neutral nature by about 2.3 times, ketone-dinitrophenolhydrozone of a basic nature - by 1.77 times, and a neutral one - by 2.5 times. From the ratio of accumulated ADNPH and KDNPH, it follows that the intake of polysaccharides led to a decrease in the accumulation of ADNPH, which are the primary markers of oxidative stress, indicating the degree of fragmentation of protein molecules. A less effective decrease in the level of KDNPH of the main nature against the background of PS intake may indicate a possible mechanism of the protective effect of PS against neutral amino acids that make up protein molecules. These data indicate a greater resistance of the heart muscle to hypoxia and the effect of modulators such as polysaccharides compared with skeletal muscle tissue. Perhaps this is due to a more powerful activation of the antioxidant system, as indicated by a change in the activity of SOD. Moreover, the degree of damage to proteins was significantly reduced when prescribing PS due to a decrease in the accumulation of ADNPH derivatives and to a lesser extent due to a decrease in KDNPH derivatives, which indicates an increase in the resistance of tissue proteins to hypoxia. Conclusions. The effect of a single hypoxia with hypercapnia provoked an increase in the activity of superoxide dismutase in the tissues of the cardiac and skeletal muscles, while SOD of the heart showed itself more actively. The intake of polysaccharides restrained the increase in SOD activity during hypoxia, however, PS had a protective effect on the proteins of skeletal muscle tissue, which led to a decrease in their oxidative modification.
Problems of Biological Medical and Pharmaceutical Chemistry. 2020;23(2):31-36
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Hepatoprotective effect of the extract of agrimony ordinary tetrachloromethane in experimental hepatitis

Kurmanova E.N., Strelkova L.B., Ferubko E.V., Sheichenko O.P.

Abstract

Relevance. The search for sources of new effective biologically active substances from plant raw materials for the creation of drugs of rehabilitation and rehabilitation orientation for the treatment of disorders in the liver is currently relevant. A promising object is the common repeche (Agrimonia eupatoria L.), which is used in folk medicine for the treatment of liver and gall bladder diseases. In GNU VILAR developed repeshko ordinary herb dry extract. The aim of the study was to study the effect of repeshka extract on the liver condition of rats under experimental toxic hepatitis. Material and methods. The pharmacological properties of the extract were studied at doses of 50 mg/kg and 100 mg/kg, the comparison drug was silimar at a dose of 100 mg/kg. The drugs were administered to animals for 3 days. Experimental hepatitis was caused by a single subcutaneous injection of carbon tetrachloride to rats one hour after the last injection of the studied substances. After 48 hours, the rats were euthanized in a CO2 chamber and the liver was extracted, from which microsomal fractions were isolated, the content of microsomal protein and cytochrome P450 was determined, as well as the rate of monooxygenase reactions catalyzed by cytochrome P450, reflecting the activity of the detoxifying microsomal system of the liver. Taking into account all the studied parameters, the hepatoprotective and detoxifying properties of the extract and the comparison drug were evaluated. Results. In the experiment it was found that in the treatment of hepatitis agrimony extract in doses of 50 mg/kg and 100 mg/kg normalizes the content of microsomal protein and cytochrome P450 content in liver microsomes increased by 28% and 2 times, respectively, compared to control. The introduction of silymar also normalizes the content of microsomal protein, and the content of cytochrome P450 increases by 16%. It was found that when treating hepatitis with repeshka extract at a dose of 50 mg/kg and 100 mg/kg, the anilization rate increases by 12% and 30%, and the demethylation rate increases by 40% and 58%, respectively. This indicates enzymatic activity centres demethylation and cytochrome P450 inducing effects of biologically active substances contained in the extract of agrimony on monooxygenase system of the liver. Silimar also showed high hydroxylase and demethylase activity. Conclusions. Thus, the dry extract of the grass repeshka ordinary in conditions of Tetrachloromethane hepatitis in rats has hepatoprotective and detoxifying properties.
Problems of Biological Medical and Pharmaceutical Chemistry. 2020;23(2):37-41
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The influence of chronic alcoholization on the functional and metabolic relations of the liver and muscles and the model of their correction

Mikashinovich Z.I., Romashenko A.V., Uspenskaya E.V.

Abstract

An analysis of rearrangements in organs with different functional specializations (liver, muscles) can clarify the nature of changes in interorgan relationships in the early stages of chronic alcoholization, which will help determine the paths of corrective actions aimed at maintaining and consolidating adaptive reactions. The aim of the study was to analyze and evaluate the nature of oxygen-dependent metabolic changes in the liver and muscles, as well as to clarify the possibility of using the drug "Tykveol" to optimize intracellular adaptive reactions in the process of alcohol intoxication. Material and Methods. The object of the study was nonlinear white rats of both sexes. The object of the study was 5 groups of nonlinear rats of both sexes. 1-3 group received 15% solution of alcohol as the only source of drinking for 3 months. Group 4 and 5 received the drug Tykveol against the background of three-month alcoholization. In the liver and cardiomyocytes, the activity of antioxidant enzymes (glutathione peroxidase, catalase) and the level of grutathione were determined. Oxygen-dependent processes were judged by the content of lactate and pyruvate in the liver and muscles. Results. When analyzing morphological changes in the pancreas by the end of 2 months of alcoholization, signs of the development of acute pancreatitis are recorded, and a histological examination after 3 months revealed karyopyknosis and karyorexis. In the process of 3-month alcoholization, an increase in the level of lactate, pyruvate and glutathione compared with the control is recorded earlier in the liver than in the muscles. Glutathione peroxidase and catalase in the early stages are activated more in the muscles than in the liver. After taking Tykveol, both the muscles and the liver stimulate the work of antioxidant enzymes, especially catalase in the liver, the level of glutathione and lactate decreases, while the content of pyruvate, especially in the muscles, exceeds the initial level. Conclusions. An analysis of the data obtained shows that under experimental conditions of alcoholization, metabolic shifts develop rapidly, and shifts of oxygen-dependent processes in the liver occur earlier than in muscle tissue. The use of Tykveol is promising in the early stages of the pathological process and is aimed at stabilizing the antioxidant system. It is necessary to take into account the identified organ-specific features associated with the accumulation of pyruvate in the muscles, which justifies the feasibility of creating complex drugs to enhance energy metabolism in cardiomyocytes in conditions of chronic alcoholization.
Problems of Biological Medical and Pharmaceutical Chemistry. 2020;23(2):42-50
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Determination of the mineral element composition of red clover grass (Trifolium pratense L.)

Nguyen T.S., Ditkovskaya Z.R., Generalova Y.E., Kaukhova I.E., Sorokin V.V.

Abstract

Chemical element influence physiological processes in the human body. Deficiency of any of the main minerals can lead to serious metabolic disorders and jeopardize the health of the human body, so the search for new types of plant materials as valuable additional sources of macro- and microelements is an actual task. Determination of the mineral composition makes it possible to predict the efficacy and safety profile of medicinal plants. The aim. The study of the qualitative and quantitative composition of meadow red clover grass to expand the range of medicines based on this plant substance. Material and Methods. The object of this study was red clover grass (Trifolium pratense L.) of firm «Horst Company LLC», Barnaul, acquired in the pharmacy chain in 2019. The mineral composition of red clover grass was determined by the method inductively coupled plasma atomic emission spectroscopy on an Optima™ 8000 spectrometer with WinLab for ICP software (Perkin Elmer Inc., USA). Results. 11 elements with a maximum content of K, Ca, and Mg ions were found in the red clover grass as the main mineral elements of this raw material. In addition, the following toxic elements are absented in red clover grass: cadmium, lead, mercury and arsenic. Conclusions. The mineral composition of red clover grass was studied. Eleven elements were found with a maximum content of potassium, magnesium, calcium, which are necessary for the normal functioning and development of the body. The following toxic elements were found to be absent in the red clover grass: cadmium, lead, mercury and arsenic. Based on the results obtained in the development of medicines, red clover grass can also be considered as a potential source of essential mineral elements.
Problems of Biological Medical and Pharmaceutical Chemistry. 2020;23(2):51-54
pages 51-54 views

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