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.

Polysaccharides PS1 and PS2 with a yield 1–2% were obtained from the above-ground part of three species of Saussurea: S. controversa DC., S. salicifolia L. and S. frolovii Ledeb. sequential extraction with water at 25 and 70 °C. PS1 free from protein impurities. A protein that is not removed by the Sevag method is co-extracted with PS2. All polysaccharides contain residues of uronic acids, the highest content found in PS1 and PS2 from S. contrоversa. The molecular weights (Mw) of PS1 from S. controversa, S. salicifolia and S. frolovii were 448.13, 158.49, 64.03 kDa and PS2 – 101.82, 94.60, 225.42 kDa, respectively. Interspecific differences in the monosaccharide composition of polysaccharides were revealed. Galactose (Gal) and Arabinose (Ara) residues are major, and Rhamnose (Rha), Xylose (Xyl) and Mannose (Man) residues are minor components of the carbohydrate chains isolated by PS. PS1 S. salicifolia and S. frolovii and PS2 S. salicifolia do not contain endotoxins impurities and have a NO-activating effect on antigen-presenting cells (macrophages), significantly exceeding the effect of muramyl dipeptide.

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.

Plant polysaccharides have a wide range of biological activity, including an immunotropic effect, the degree and magnitude of which depends on the composition and tertiary structure of the macromolecule. Acidic polysaccharides (APS) were isolated by extraction with an acidic aqueous solution (pH=3) and characterized From the rhizomes of Acorus calamus L., the aerial part of Centaurea pseudomaculosa Dobrocz., the herb Conium maculatum L., and the leaves of Betula pendula Roth. The content of total carbohydrates was determined for all samples (>90%), uronic acids (24.56–53.70%) and a small amount of impurity substances (protein and nucleic acids 0.81–1.02% and 0.016–0.035%, respectively); molecular weight distribution (Mn, Mw). Structural features were studied using physicochemical and instrumental methods, such as UV and IR spectroscopy, reaction with Congo red, and scanning electron microscopy. The polysaccharides of Acorus calamus, Conium maculatum, and Betula pendula have a triple-helical conformation and bulky ultrastructure; the polysaccharides of Centaurea pseudomaculosa are characterized by the absence of a triple-helical conformation and an amorphous ultrastructure.

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% hexose, 12.99 ± 0.26% uronic acids, and 5.24 ± 0.04% protein and neutral monosaccharides represented by glucose, galactose, xylose, and rhamnose in a molar ratio of 1.67 : 1.00 : 1.20 : 1 > 42. Methods: Five main components were isolated by ion exchange chromatography. The structure of the isolated polysaccharides (PSs) was characterized by physicochemical methods, specifically IR spectroscopy, high-performance exclusive chromatography, and gas-liquid chromatography, as well as by the Congo red reaction. Results and Discussion: All the PSs have different contents 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 little of protein (0.75 ± 0.13 to 2.28 ± 0.17%). The isolated polysaccharide samples are highly homogeneous and have different monomeric compositions. The major component of SS-1 PS is galactose, SS-2, SS-3, and SS-4 PSs contain mostly rhamnose, and glucose and xylose predominate in SS-5 PS. The triple helix structure is established for SS PSC and SS-5 PS. Conclusions: The isolated PSs stimulated NO synthase of peritoneal macrophages at the lipopolysaccharide (LPS) level, and samples 3, 4, and 5 showed an endotoxin-independent activity.

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.