ABSTRACT
The ultrastructure of nephrocytes of the proximal and distal convoluted tubules, podocytes, mesangial cells, and macrophages of the interstitial connective tissue was studied after single intravenous administration of magnetite nanoparticles modified with chitosan (magnetic nanospheres) or lipids (magnetic liposomes). Transmission electron microscopy showed ultrastructural features of absorption of magnetite nanoparticles. The shape, size, and number of vesicles containing nanoparticles in nephrocytes of convoluted tubules and macrophages after administration of the suspensions of magnetic nanospheres and magnetic liposomes were described.
Subject(s)
Macrophages/drug effects , Magnetite Nanoparticles/administration & dosage , Mesangial Cells/drug effects , Podocytes/drug effects , Administration, Intravenous , Animals , Animals, Outbred Strains , Chitosan/chemistry , Endocytosis , Liposomes/administration & dosage , Macrophages/metabolism , Macrophages/ultrastructure , Male , Mesangial Cells/metabolism , Mesangial Cells/ultrastructure , Podocytes/metabolism , Podocytes/ultrastructure , RatsABSTRACT
We studied activation of macrophages with humic acids extracted from peat of large deposits in the Tomsk region by two extraction methods: by hydroxide or sodium pyrophosphate. Humic acid of lowland peat types containing large amounts of aromatic carbon, phenolic and alcohol groups, carbohydrate residues and ethers, irrespectively of the extraction methods contained LPS admixture that probably determines their activating properties. Humic acid of upland peat types characterized by high content of carbonyl, carboxyl, and ester groups enhance NO production and reduce arginase expression, but these effects were minimized when sodium hydroxide was used as an extraction solvent. Pyrophosphate samples of the upland peat types were characterized by aromaticity and diversity of functional groups and have a significant advantage because of they induce specific endotoxin-independent stimulating action on antigen presenting cells.
Subject(s)
Complex Mixtures/pharmacology , Humic Substances/analysis , Macrophages, Peritoneal/drug effects , Soil/chemistry , Animals , Arginase/metabolism , Carbon/analysis , Complex Mixtures/isolation & purification , Diphosphates/analysis , Diphosphates/chemistry , Female , Lipopolysaccharides/pharmacology , Liquid-Liquid Extraction/methods , Macrophages, Peritoneal/cytology , Macrophages, Peritoneal/immunology , Male , Mice , Mice, Inbred C57BL , Nitric Oxide/biosynthesis , Phenols/analysis , Polymyxin B/pharmacology , Primary Cell Culture , Sodium Hydroxide/chemistryABSTRACT
A screening study of biological activity of native humic acids isolated from peat was performed; several physical and chemical parameters of their structures were studied by UV- and infrared spectroscopy. Spectroscopy yielded similar shape of light absorption curves of humic acids of different origin, which can reflect similarity of general structural principles of these substances. Alkaline humic acids have more developed system of polyconjugation, while molecular structures of pyrophosphate humic acids were characterized by higher aromaticity and condensation indexes. Biological activity of the studied humic acids was assessed by NO-stimulating capacity during their culturing with murine peritoneal macrophages in a wide concentration range. It was shown that due to dose-dependent enhancement of NO production humic acids can change the functional state of macrophages towards development of pro-inflammatory properties. These changes were associated with high activity of humic acids isolated by pyrophosphate extraction, which allows considering effects of isolation method on biological activity.
Subject(s)
Humic Substances , Macrophages, Peritoneal/metabolism , Nitric Oxide/biosynthesis , Soil/chemistry , Animals , Cells, Cultured , Drug Evaluation, Preclinical , Female , Macrophages, Peritoneal/drug effects , Male , Mice, Inbred C57BLABSTRACT
Screening study of the effects of sweet flag (Acorus calamus L.) rhizome and clover (Trifolium pratense L.) aerial part on the production of NO by mouse macrophages was carried out. The polysaccharides were separated by ion exchange chromatography into fractions differing by monomeric composition and ramification type and were used in concentrations of 20, 40, and 100 µg/ml. Four fractions of Acorus calamus L. (PSF-101, PSF-102, PSF-103, and PSF-105), used in different concentrations, moderately stimulated nitrite production by macrophages. Three of five Trifolium pratense L. polysaccharides (PS62-3, PS62-4, and PS62-5) exhibited a significant specific effect on NO production. Rhamnogalactouronans from clover PS63-3 in all concentrations and from PS62-5 in a concentration of 100 µg/ml exhibited the highest activity, comparable to the NO-stimulatory activity of the reference LPS, while polysaccharide PS62-3 in a concentration of 40 µg/ml exhibited even higher activity.
Subject(s)
Acorus/chemistry , Macrophages/drug effects , Macrophages/metabolism , Nitric Oxide/metabolism , Pectins/pharmacology , Polysaccharides/pharmacology , Trifolium/chemistry , Animals , Cells, Cultured , Female , Male , Mice , Pectins/chemistry , Polysaccharides/chemistryABSTRACT
We studied the effect of two pectic polysaccharides PS-B1-AG and PS-B2-RG that were contained in total polysaccharides extracted from Betula pendula leaves on NO production by mouse macrophages and the contribution of signaling molecules to macrophage activation by the test substances. Unlike the total sample, pectins produced a NO-stimulating effect on macrophages. The effect of PS-B2-RG (10 µg/ml) did not differ from the effect of LPS, while PS-B1-AG produced this effect only in a concentration of 20 µg/ml, which was probably due to differences in the chemical structure of the test substances. The studied pectin polysaccharides activated transcription factor NF-κB, kinases p38 and PI3, and cAMP as a negative regulator. These results indicate that Betula pendula polysaccharides are promising substances for creation of immunomodulating drugs.
Subject(s)
Betula/chemistry , Immunologic Factors/pharmacology , Macrophage Activation/drug effects , Polysaccharides/pharmacology , Animals , Cells, Cultured , Cyclic AMP/metabolism , Female , Lipopolysaccharides/pharmacology , Macrophages/immunology , Macrophages/metabolism , Male , Mice, Inbred BALB C , Mice, Inbred C57BL , NF-kappa B/metabolism , Nitric Oxide/biosynthesis , Phosphatidylinositol 3-Kinases/metabolism , Plant Leaves/chemistry , Signal Transduction , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
A pectic polysaccharide was isolated from the rhizomes of Acorus calamus L. The main component of its carbohydrate chain was represented by residues of D-galacturonic acid (>85%). In addition, this polysaccharide contained residues of galactose, arabinose, xylose and rhamnose (<10%). Structural analysis of pectic polysaccharide from A.calamus L. with NMR spectroscopy indicated that it contains the regions of a linear 1,4-alpha-D-galactopyranosyluronan, which represents a major component of the macromolecule. A considerable amount of galacturonic acid residues was not methoxylated. We demonstrate here that the pectic polysaccharide from A. calamus L. in low concentrations was able to stimulate in vitro IL-12 and nitric oxide production by murine macrophages. It also induced TNF-alpha secretion by human peripheral blood mononuclear cells, reduced arginase activity but did not affect IL-10 secretion by murine macrophages or human peripheral blood mononuclear cells. The IL-12 and NO-stimulating effects on murine macrophages were similar to that of LPS. In addition, the polysaccharide promoted in vivo Th1 immune response in mice which were immunized with sheep red blood cells (DTH and quantity of plaque-forming cells) and down regulated serum level of IgG1 and IgE during Th2-depend immune response induced by ovalbumin. The PS increased Th1-induced edema and suppressed Th2-induced paw swelling in adoptive systems. Our results suggest that the pectic polysaccharide from A. calamus L. represents a promising immunomodulating agent that stimulates M1-polarized macrophages and promotes Th1-oriented adaptive immune response. We propose that this polysaccharide could be potentially applied for treatment of infectious, oncological diseases or for immunoglobulin-E-mediated disorders.
