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.

This comprehensive review presents the results of an in-depth analytical literature search on the biological activity of humic substances and their possible pharmacological mechanisms of action. The unique chemical structure of humic substances has determined their widespread use in many economic sectors, including medicine. Thanks to modern advances in pharmaceuticals, pharmacology, and toxicology, it has been possible to demonstrate the multifaceted biological activity of humic substances and, consequently, the possibility of using them to treat and prevent many infectious and non-infectious pathologies, including diseases considered incurable. The article presents data on their immunotropic, antibacterial, antiviral (including HIV), antitumor, antioxidant and antiradical, cardiotropic, hepatoprotective, regenerative, detoxifying, and adaptogenic effects; their influence on the intestinal microbiome; studies of the toxic properties of humic substances and the safety of their use in medicine; and the current trend of using humic substances as unique matrices for creating next-generation bionanomaterials. An analysis of data on the intracellular mechanisms that play a key role in the implementation of the effects of humic substances is conducted. Thus, the natural genesis of humic substances, their multifaceted biological activity, and the absence of toxic and allergenic properties explain the growing interest of scientists from all over the world in their study.

Substances of silver nanoparticles dialyzed through a 13 kDa membrane, synthesized in a medium of humic ligands modified with hydroquinone and 2-hydroxynaphthoquinone from PowHumus brown coal, specifically enhance the M2 properties of peritoneal macrophages due to inhibition of NO synthase and significant activation of arginase, thus enhancing anti-inflammatory properties of cells. In small, but effective concentrations, they do not have cytotoxic properties and do not contain pyrogenic impurities. The studied humates are able to influence the mechanisms of immune response formation and are an effective means for correcting inflammation and regeneration.

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.

We performed a comparative in vitro study of the involvement of NF-κB, PI3K, cAMP, ERK1/2, p38, JAKs, STAT3, JNK, and p53-dependent intracellular signaling in the functioning of neural stem cells (NSC) under the influence of basic fibroblast growth factor (FGF) and FGF receptor agonist, diterpene alkaloid songorine. The significant differences in FGFR-mediated intracellular signaling in NSC were revealed for these ligands. In both cases, stimulation of progenitor cell proliferation occurs with the participation of NF-κB, PI3K, ERK1/2, JAKs, and STAT3, while JNK and p53, on the contrary, inhibit cell cycle progression. However, under the influence of songorin, cAMP- and p38-mediated cascades are additionally involved in the transmission of the NSC division-activating signal. In addition, unlike FGF, the alkaloid stimulates progenitor cell differentiation by activating ERK1/2, p38, JNK, p53, and STAT3.

An investigation was carried out on humic substances (HSs) isolated from the coal of the Kansk-Achinsk basin (Krasnoyarsk Territory, Russia). The coal HSs demonstrate the main parameters of molecular structure inherent to this class of natural compounds. An assessment was performed for the chemical, microbiological, and pharmacological safety parameters, as well as the biological efficacy. The HS sample meets the safety requirements in microbiological purity, toxic metals content (lead, cadmium, mercury, arsenic), and radionuclides. The presence of 11 essential elements was determined. The absence of general, systemic toxicity, cytotoxicity, and allergenic properties was demonstrated. The coal HS sample was classified as a Class V hazard (low danger substances). High antioxidant and antiradical activities and immunotropic and cytoprotective properties were identified. The ability of the HS to inhibit hydroxyl radicals and superoxide anion radicals was revealed. Pronounced actoprotective and nootropic activities were also demonstrated in vivo. Intragastric administration of the HS sample resulted in the improvement of physical parameters in mice as assessed by the “swim exhaustion” test. Furthermore, intragastric administration in mice with cholinergic dysfunction led to a higher ability of animals with scopolamine-induced amnesia to form conditioned reflexes. These findings suggest that the studied HS sample is a safe and effective natural substance, making it suitable for use as a dietary bioactive supplement.

Objective: A polysaccharide complex (PSC) from the aboveground part of Saussurea salicifolia L. (SS) was isolated and studied to show that SS PSC contains 54.99 ± 0.53

The effect of polysaccharides isolated from the aboveground parts of Saussurea salicifolia (L.) DC on Th2 type immune response reactions was studied. Administration of water-soluble polysaccharides presented by arabino-galacturonans (weight average molecular weight 158.49 kDa) to mice against the background of experimental Th2 immunity reduced the severity of anaphylactic and local immediate type hypersensitivity reactions. It also suppressed the production of ovalbumin-specific IgE and IgG1 and increased the stability of mast cell membranes. The studied polysaccharide complex increased IFNγ secretion and inhibited IL-4 synthesis. These findings suggest that these polysaccharides may be considered as potential anti-allergic agents that suppress the development of allergy in its early stages.

Humic acids isolated by sodium pyrophosphate extraction from various types of peat activate macrophages in the classical proinflammatory pathway and stimulate nitric oxide production by these cells. This effect is mediated by activation of intracellular signaling pathways involving MAPK p38, PI3K, MEK 1/2 kinase, cAMP, and NF-κB via TLR-2 and TLR-4 receptors.

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.

Physicochemical and pharmacological properties of the humic substances from coal of the Kansk-Achinsk coal basin, the Pereyaslovskoe deposit, have been studied. The presence of polycyclic aromatic structures and aliphatic fragments with various functional groups as substituents (carboxyl, carbonyl and quinoid, phenolic, alcoholic, ether and ester, amino and amide) was established by means of UV, IR, 13C NMR spectroscopy, fluor-imetry, and elemental analysis. It has been determined by HPLC that coal humic substances are highly hydro-philic polydisperse biopolymers with medium molecular weights. According to the indicator of microbiological purity (category 3B), the content of microorganisms does not exceed the indicators required by the State Pharmacopeia of the Russian Federation (SPh XIV edition). The content of radionuclides, toxic metals (lead, cadmium, mercury, arsenic) corresponds to the norms (according to SPh XIV edition). The presence of 11 essential elements has been established. The studied humic substances belong to the V class of hazard (low-hazard sub-stances, according to GOST 32644-2014), do not have allergising properties and cytotoxic effect within a broad concentration range. Antioxidant, immunotropic, cytoprotective properties were established in the studies of specific pharmacological activity with various experimental models. The ability to inhibit free radicals such as hydroxyl radicals (HO.) and superoxide-anion radical (O2-. should be stressed, as these radicals are able to by-pass the endogenous antioxidant defense systems of the body. It is concluded that the studied humic substances of the Kansk-Achinsk coal basin, the Pereyaslovskoe deposit, are safe and effective biologically active sub-stances of natural origin for use as a promising biologically active substance in the food and pharmaceutical industries.

Drug development & registration. 2022;11(2):59–64. (In Russ.) https://doi.org/10.33380/2305-2066-2022-11-2-59-64. Published: 25.05.2022.

The course administration of humic acids isolated with sodium pyrophosphate from raised bog sphagnum peat reduces the parameters of a Th1-type immune response in C57BL/6 mice, the severity of an anaphylactic shock in outbred CD1 mice, and degranulation of mast cells after their immunization with ovalbumin. The addition of humic acids increases the stimulated production of IL-4, IL-10 and reduces the production of IL-2, IFNγ by peripheral blood mononuclear cells of healthy donors.

We studied the involvement of cAMP and PKA in the regulation of the secretion of neurotrophic growth factors by macro-and microglial cells in the model of ethanol-induced neurodegeneration in vitro and in vivo . The stimulating role of cAMP in the secretion of neurotrophins by intact astrocytes and oligodendrocytes was shown, while PKA does not participate in this process. On the contrary, the inhibitory role of cAMP (implemented via PKA activation) in the production of neurogenesis stimulators by microglial cells under conditions of optimal vital activity was found. Under the influence of ethanol, the role of cAMP and PKA in the production of growth factors by macroglial cells was considerably changed. The involvement of PKA in the cAMP-dependent signaling pathways and inversion of the role of this signaling pathway in the implementation of the neurotrophic secretory function of astrocytes and oligodendrocytes, respectively, directly exposed to ethanol in vitro were noted. Long-term exposure of the nervous tissue to ethanol in vivo led to the loss of the stimulating role of cAMP/PKA signaling on neurotrophin secretion by macroglial cells without affecting its inhibitory role in the regulation of this function in microglial cells.

The course administration of humic acids isolated by the sodium pyrophosphate method from pine-sphagnum-cotton sedge peat reduced the general anaphylaxis reaction in mice and guinea pigs immunized with ovalbumin and decreased serum content of IgG1 and IgE in mice. The serum from mice treated with humic acids and sensitized with ovalbumin did not increase the rate of degranulation of mast cells isolated from intact Wistar rats in the presence of ovalbumin in comparison with the serum of control animals.

The effects of JAK and STAT3 inhibitors on the production of neurotrophic growth factors by different types of neuroglial cells were studied under conditions of in vitro and in vivo models of ethanol-induced neurodegeneration. It was shown that these signaling molecules do not participate in the secretion of neurotrophins by intact astrocytes and oligodendrocytes. The inhibitory role of JAK in the regulation of this function of microglial cells was revealed. We also revealed significant changes in the role of JAK and the presence of STAT3 specifics within the framework of JAK/STAT signaling in the production of growth factors by various glial elements under the influence of ethanol. Neurodegeneration modeled in vitro led to the appearance of a “negative” effect of STAT3 on the production of neurogenesis stimulants by all types of glial cells. Moreover, the role of STAT3 in oligodendrocytes and microglial cells generally corresponded to that of JAK/STAT signaling. In astrocytes, only selective blockade of STAT3 (but not JAK) led to stimulation of their function. In mice subjected to prolonged peroral alcoholization, the neuroglial responses to the pharmacological regulation of JAK/STAT signaling were different. An inversion of the role of JAK and STAT3 in the production of neurotrophins by oligodendrocytes was noted. In addition, JAK inhibitor did not stimulate secretory function of microglial cells under conditions of prolonged exposure to ethanol in vivo.

Introduction. The nitric oxide molecule is one of the most versatile, multifunctional mediators of biological reactions in the body. NO participates in the formation of innate and specific immunity, the inflammatory process and protection against infectious pathogens. It is known that polysaccharides of higher plants are widely used to activate pro- and anti-inflammatory reactions. Previous studies have shown the promise of a more detailed research in the immunomodulatory properties of plant polysaccharides of the genus Saussurea DC, the mechanisms of their immunostimulating and anti-inflammatory action depending on the origin and conditions of extraction. Aim. The aim of present research was to evaluate the dependence of the affection of the extraction parameters of water-soluble polysaccharides (WSPS) from the aerial parts of S. salicifolia (L.) DC., S. controversa DС., S. frolovii Lebeb. on NO synthase activity and peritoneal macrophages proliferation. Materials and methods. Chemical methods for the isolation of water-soluble polysaccharides, cultural and biochemical methods for assessing biological activity were used. Results and discussion. Water-soluble polysaccharides (WSPS) from plants S. salicifolia (L.) DC., S. controversa DС., S. frolovii Ledeb. obtained with different extraction parameters have the ability to enhance production of nitrites antigen-presenting cells in the same way as standard macrophage activator liposaccharide. Polysaccharides from S. controversa and S. frolovii at acidic extractant are free of endotoxin impurities. All water-soluble polysaccharide samples obtained using neutral and alkaline extractants contain endotoxin impurities. Samples from the S. controversa contained this impurity after extraction in an acidic extractant. Polysaccharide samples are not cytotoxic. Conclusion. It was stated that water-soluble polysaccharides from plants of the Saussurea genus using an acidic extractant do not possess cytotoxic properties, do not contain endotoxin impurities and significantly enhance the production of nitric oxide by macrophages. These samples used in a detailed study of the immunomodulatory properties and further development on their basis of new treatment methods various diseases that require correction and regulation of the functional activity of immune cells, including macrophages.

Introduction . Macrophages occupy a special place in the formation and coordination of the immune response, their population is characterized by heterogeneity, however, two main types are distinguished among them: pro-inflammatory (classically activated) macrophages M1, which are activated by interferon-γ (IFN-γ) or lipopolysaccharide (LPS) of bacteria, produce pro-inflammatory cytokines, nitric oxide (NO) and initiate an immune response, and anti-inflammatory (alternatively activated) macrophages M2, which are activated by such cytokines as interleukin 1 (IL-1), IL-10 or IL-13 and are associated with suppression of the immune response and tissue healing. The proinflammatory properties of macrophages are most often associated with the activation of an intracellular signaling cascade with the participation of the IKK-2 kinase complex and nuclear factor-κB (NF-κB), which is the most important signal transduction molecule. Previously, the authors showed that humic acids (HA) of peat cause the appearance of pro-inflammatory properties in macrophages; however, the intracellular mechanisms of such activation have not been studied. Aim . Study of the contribution of the intracellular signaling cascade IKK-2/NF-κB to the activation of pro-inflammatory properties of macrophages by humic acids of peat. Materials and methods . The studies were carried out using cultural methods and western blotting. Results and discussion . It was shown that the addition of the IKK-2 inhibitor to the macrophage culture reduced the ability of HA to stimulate the production of nitric oxide by the cells. Western blotting revealed that after 30 minutes simultaneous incubation of HA with macrophages, the content of the NF-κB p65 molecule in the cells increased, after 60 minutes it decreased to the values of intact control, and after 17 hours it decreased 3 times compared to the control level. However, the IKK-2 inhibitor after a 30-minutes incubation did not have a blocking effect on the IKK kinase, whereas in an hour of the co-incubation of the cells with the inhibitor and substances, a decrease in the level of NF-κB p65 was observed. This trend continued in 17 hours. Conclusion. The stimulating effect of peat humic acids on macrophages is realized, among other things, due to the activation of the IKK-2 kinase complex with further release of the p65 subunit of the NF-κB molecule. The dependence of the degree of NF-κB activation on the period of interaction of the cell with activating agents is traced. HA start the intracellular signaling cascade of the nuclear factor NF-κB already within the first 30 minutes.

Introduction. The nitric oxide molecule is one of the most versatile, multifunctional mediators of biological reactions in the body. NO participates in the formation of innate and specific immunity, the inflammatory process and protection against infectious pathogens. It is known that polysaccharides of higher plants are widely used to activate pro- and anti-inflammatory reactions. Previous studies have shown the promise of a more detailed research in the immunomodulatory properties of plant polysaccharides of the genus Saussurea DC, the mechanisms of their immunostimulating and anti-inflammatory action depending on the origin and conditions of extraction.Aim. The aim of present research was to evaluate the dependence of the affection of the extraction parameters of water-soluble polysaccharides (WSPS) from the aerial parts of S. salicifolia (L.) DC., S. controversa DС., S. frolovii Lebeb. on NO synthase activity and peritoneal macrophages proliferation.Materials and methods. Chemical methods for the isolation of water-soluble polysaccharides, cultural and biochemical methods for assessing biological activity were used.Results and discussion. Water-soluble polysaccharides (WSPS) from plants S. salicifolia (L.) DC., S. controversa DС., S. frolovii Ledeb. obtained with different extraction parameters have the ability to enhance production of nitrites antigen-presenting cells in the same way as standard macrophage activator liposaccharide. Polysaccharides from S. controversa and S. frolovii at acidic extractant are free of endotoxin impurities. All water-soluble polysaccharide samples obtained using neutral and alkaline extractants contain endotoxin impurities. Samples from the S. controversa contained this impurity after extraction in an acidic extractant. Polysaccharide samples are not cytotoxic.Conclusion. It was stated that water-soluble polysaccharides from plants of the Saussurea genus using an acidic extractant do not possess cytotoxic properties, do not contain endotoxin impurities and significantly enhance the production of nitric oxide by macrophages. These samples used in a detailed study of the immunomodulatory properties and further development on their basis of new treatment methods various diseases that require correction and regulation of the functional activity of immune cells, including macrophages.