The genus Saussurea has been used in the preparation of therapies for a number of medical problems, yet not much is known about the therapeutic high-molecular-weight compounds present in extracts from these plants. Since polysaccharides are important in immune modulation, we investigated the chemical composition and immunomodulatory activity of Saussurea salicifolia L. and Saussurea frolovii Ledeb polysaccharides. Water-soluble polysaccharides from the aerial parts of these plants were extracted using water at pHs of 2 and 6 and subsequently precipitated in ethanol to obtain fractions SSP2 and SSP6 from S. salicifolia and fractions SSF2 and SSF6 from S. frolovii. The molecular weights of fractions SSP2, SSP6, SFP2, and SFP6 were estimated to be 143.7, 113.2, 75.3, and 64.3 kDa, respectively. The polysaccharides from S. frolovii contained xylose (67.1–71.7%) and glucose (28.3–32.9%), whereas the polysaccharides from S. frolovii contained xylose (63.1–76.7%), glucose (11.8–19.2%), galactose (4.7–8.3%), and rhamnose (6.8–9.4%). Fractions SSP2, SSP6, and SFP2 stimulated nitric oxide (NO) production by murine macrophages, and NO production induced by SSP2, SSP6, and SFP2 was not inhibited by polymyxin B treatment of the fractions, whereaspolymyxin B treatment diminished the effects of SFP6, suggesting that SFP6 could contain lipopolysaccharide (LPS). The LPS-free fractions SSP2, SSP6, and SFP2 had potent immunomodulatory activity, induced NO production, and activated transcription factors NF-κB/AP-1 in human monocytic THP-1 cells and cytokine production by human MonoMac-6 monocytic cells, including interleukin (IL)-1α, IL-1β, IL-6, granulocyte macrophage colony-stimulating factor (GM-CSF), interferon-γ, monocyte chemotactic protein 1 (MCP-1), and tumor necrosis factor (TNF). These data suggest that at least part of the beneficial therapeutic effects reported for water extracts of the Saussurea species are due to the modulation of leukocyte functions by polysaccharides.

A polysaccharide complex (PSC) from the aboveground part of Saussurea salicifolia L. (SS) was isolated and studied. SS PSC contains 54.99±0.53% hexose, 12.99±0.26% uronic acids, 5.24±0.04% protein, neutral monosaccharides are represented by glucose, galactose, xylose and rhamnose (molar ratios: 1,67 : 1,00 : 1,20 : 1,42). Five main components were isolated using ion exchange chromatography. The structure of the obtained polysaccharides (PS) 1-5 was characterized using physicochemical methods, such as IR spectroscopy, high-efficiency exclusive and gas-liquid chromatography, reactions with congo red. All PS are characterized by a different content of hexoses (from 32.46±2.63 to 83.64± 5.93%), uronic acids (from 2.56± 0.17 to 20.07± 3.70%) and a slight admixture of protein (from 0.75±0.13 to 2.28± 0.17%). The polysaccharides obtained are highly homogeneous samples, different in monomeric composition. The major component of SS-1 PS is represented by galactose, SS-2, 3, 4 PS – rhamnose, and SS-5 PS – glucose and xylose. The triple helix structure is installed for SS PSC and PS SS-5. All isolated PS stimulated NO synthase of peritoneal macrophages at control level 2 (LPS), and samples 3, 4 and 5 showed endotoxin-independent activity.

Изучено влияние индивидуальных компонентов экстракта соссюреи спорной (Saussurea controversa DC) на пролиферацию и NO-продуцирующую активность перитонеальных макрофагов мышей. Из бутанольной и водной фракций экстракта S. controversa изолированы кверцетин-3-О-β-D-диглюкозид-О-α-L-рамнозид (SC-1), кверцетин-3-рутинозид (рутин) (SC-2), кверцетин-7-О-α-L-рамнозид-3-О-β-D-ксилозид (SC-3), кверцетин-7-О-β-D-глюкозид-3-О-α-L- рамнозид-О-β-D-глюкозид (SC-4), хелидоновая кислота и композиция, содержащая хлорогеновую кислоту и аминокислоты. NO-стимулирующее влияние на макрофаги проявили SC-3 (80 мкг/мл), SC-4 (20 и 80 мкг/мл) и хелидоновая кислота (5 и 20 мкг/мл). Рутин в концентрации 80 мкг/мл, напротив, способствовал снижению концентрации NO. SC-3, SC-4 и хелидоновую кислоту можно рассматривать в качестве перспективных фармакологических веществ для профилактики и терапии хронических воспалительных заболеваний, SC-2 — для терапии заболеваний, сопровождающихся чрезмерной воспалительной реакцией. Исследуемые соединения требуют дальнейшего углубленного изучения с перспективой применения для регуляции/управления воспалительным процессом, переключения фенотипа антигенпрезентирующих клеток при сочетанной терапии заболеваний, сопровождающихся реактивной воспалительной реакцией.

Introduction. Representatives of the Ericaceae family are quite common in Russia and are promising for the creation of new medicines of plant origin. At the same time, only 4 of them are official. The study of biologically active substances and pharmacological activity of Andromeda polifolia L., Chamaеdaphne calyculata (L.) Moench, Ledum palustre L., Empetrum nigrum L. with rich resource reserves is promising.Aim. Comparative study of the composition of total flavonoid fractions from A. polifolia, C. calyculata, L. palustre, E. nigrum and study of their effect on the NO-stimulating activity of peritoneal macrophagesMaterials and methods. The crushed aboveground part (leafy shoots) was previously depigmented with chloroform, treated with 70 % aqueous acetone, then acetone was removed. Flavonoids were extracted with ethyl acetate from the aqueous phase. The identification of flavonoids was carried out by HPLC (UltiMate 3000 chromatograph) according to the coincidence of retention times and spectral characteristics, the calculation of the content was carried out by simple normalization. The effect of the samples on nitric oxide production was studied on macrophages of C57BL/6 mice. Endotoxin control in the samples was carried out using a LAL test and cell incubation in the presence of polymyxin B.Results and discussion. The composition of flavonoid fractions from A. polifolia, C. calyculata, L. palustre and E. nigrum has been studied. 8 phenolic compounds were found in C. calyculata shoots, including isoquercitrin, herbacetin, naringenin and naringin – for the first time for this species. 5 compounds were detected in A. polifolia shoots, including isoquercitrin and herbacetin, for the first time for this species. 5 and 4 compounds were identified in L. palustre and E. nigrum shoots, respectively, while quercetin glycosides are predominant in all samples: isoquercitrin, hyperoside and rutin. The fraction of C. calyculata flavonoids at doses of 1, 5, 10 mcg/ml inhibits the production of nitric oxide by macrophages by 30 %, and E. nigrum flavonoids at doses of 100 and 200 mcg/ml, on the contrary, enhance the production of nitrites by macrophages by 33 and 37 %, respectively.Conclusion. A comparative study of the composition of total flavonoid fractions from A. polifolia, C. calyculata, L. palustre, and E. nigrum, which are capable of activating both M1 and M2 polarization of peritoneal macrophages in mice, has been conducted, which requires further in-depth study. The flavonoids C. calyculata and E. nigrum are promising for further study.

It has recently been shown that by sequential vibrational processing (named ‘crossing’) of two closely located vials with the initial substance in one of them and water or lactose in the other, it is possible to obtain novel products, or vibrational iterations. Vibrational iterations are, in certain cases, capable of exerting a modifying effect on the initial substance or its target, thereby altering its physicochemical and biological properties. Since water is the environment in which metabolic processes occur in cells, organs, and tissues, vibrational iterations obtained using water as initial substance can potentially affect their course. In this work, we assessed the effect of vibrational iterations of water on the metabolic activity of primary peritoneal macrophages of C57BL/6 mice and macrophage-like tumor cells of the RAW 264.7 line. It was shown that one vibrational iteration of water increased the metabolic activity of peritoneal macrophages and suppressed the metabolic activity of tumor cells. At the same time, another vibrational iteration of water did not influence the metabolic activity of peritoneal macrophages but decreased the metabolic activity of tumor cells. Probably, taking into account the modifying activity of vibrational iterations, their action is aimed at changing the properties (possibly, structuring) of intracellular water. The results obtained by us show the possibility of a fundamentally new differential effect on the metabolic activity of healthy and tumor cells, which opens up new prospects for antitumor therapy.

Aim. To study the effect of a course administration of peat humic acids from bogs of the Tomsk Region on the phagocytic activity of peritoneal macrophages and blood neutrophils in mice, and the thymus-dependent humoral immune response induced by the administration of sheep erythrocytes. Materials and methods. The following immunological methods were used: study of the phagocytic activity of peritoneal macrophages and blood neutrophils of mice, determination of the number of antibody-forming cells (AFC) in the spleen of mice, measurement of the serum level of antibodies to sheep erythrocytes by the hemagglutination assay after the course administration of three samples of peat humic acids. The experiment involved 70 C57Bl/6J mice (females), aged 7–8 weeks. Results. It was found that the course administration of all studied samples of humic acids resulted in an increase in the number of phagocytes and the intensity of particle engulfment. Two out of three samples enhanced the humoral immune response induced by the administration of sheep erythrocytes, which was manifested by an increase in the number of AFCs in the spleen and the hemagglutination assay titer in the blood serum. Conclusion. The samples of humic acids that influence the phagocytic function of macrophages and neutrophils and enhance the humoral immune response may serve as a basis for the development of new therapeutic agents for the treatment of immunodeficiency states.