ABSTRACT
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.
Subject(s)
Arginase , Arginine , Humic Substances , Macrophages, Peritoneal , Silver , Animals , Macrophages, Peritoneal/drug effects , Macrophages, Peritoneal/metabolism , Mice , Arginine/pharmacology , Arginine/chemistry , Arginase/metabolism , Silver/chemistry , Silver/pharmacology , Metal Nanoparticles/chemistry , Hydroquinones/pharmacology , Hydroquinones/chemistry , Male , Nitric Oxide/metabolism , Nitric Oxide Synthase/metabolism , Naphthoquinones/pharmacology , Naphthoquinones/chemistryABSTRACT
The pharmacological activity of granulocyte CSF (G-CSF) immobilized using electron-beam synthesis nanotechnology (imG-CSF) was evaluated in an experimental model of ovarian reserve depletion. The effectiveness of the drug was compared with that of its unmodified form. Depletion of the ovarian follicular pool in female Sprague-Dawley rats was caused by a single intravenous injection of the antitumor drug etoposide in the maximum tolerated dose. The effectiveness of the studied drugs was assessed by serum concentration of anti-Mullerian hormone (AMH) measured by ELISA and by the number of primordial, two-layer, multilayer, and atretic follicles counted on serial sections of the ovaries (5-µm thick; through the entire organ) stained with hematoxylin and eosin. It was found that imG-CSF prevents depletion of the ovarian reserve in the model used, which was confirmed by high AMH concentration and higher numbers of primordial, two- and multilayer follicles in comparison with the corresponding parameters in the control (etoposide), and by a decrease in the severity of atretic processes. Unmodified form of the drug demonstrated lower efficiency.
Subject(s)
Ovarian Reserve , Rats , Animals , Female , Etoposide , Granulocyte Colony-Stimulating Factor/pharmacology , Electrons , Rats, Sprague-Dawley , Anti-Mullerian Hormone , Models, TheoreticalABSTRACT
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.
Subject(s)
Humic Substances , Soil , Macrophage Activation , Nitric Oxide Synthase Type II/metabolism , Signal Transduction , NF-kappa B/metabolism , Nitric Oxide/metabolism , p38 Mitogen-Activated Protein Kinases/metabolism , Lipopolysaccharides/pharmacologyABSTRACT
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.
Subject(s)
Anti-Allergic Agents , Sphagnopsida , Animals , Anti-Allergic Agents/pharmacology , Cell Degranulation , Guinea Pigs , Humic Substances , Immunoglobulin E , Mast Cells , Mice , Ovalbumin , Rats , Rats, Wistar , SoilABSTRACT
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.
Subject(s)
Humic Substances , Sphagnopsida , Mice , Animals , Humic Substances/analysis , Soil , Leukocytes, Mononuclear/chemistry , Wetlands , Mice, Inbred C57BL , Immunity , Th2 Cells , Cytokines , Th1 CellsABSTRACT
The in vitro addition of water-soluble polysaccharides isolated from the leaves of Crataegus sanguinea Pall. to culture of mouse peritoneal macrophages induced classical activation of antigen-presenting cells by increasing NO synthase activity and reducing arginase expression.
Subject(s)
Crataegus/chemistry , Macrophages, Peritoneal/drug effects , Nitric Oxide/metabolism , Polysaccharides/pharmacology , Animals , Arginase/drug effects , Arginase/metabolism , Cells, Cultured , Female , Macrophages, Peritoneal/enzymology , Macrophages, Peritoneal/metabolism , Mice , Mice, Inbred C57BL , Plant Extracts/chemistry , Plant Extracts/pharmacology , Plant Leaves/chemistry , Polysaccharides/isolation & purification , Solubility , Water/chemistryABSTRACT
Humic acids extracted with sodium pyrophosphate from Oligotrophic Sphagnum magellanicum peat reduce mitogen-stimulated production of anti-inflammatory cytokine IL-10 by mouse peritoneal macrophages and do not affect the secretion of IL-4 by lymphocytes. The studied humic acid sample stimulates the production of proinflammatory cytokines IL-12, TNFα, IL-1ß, and IFNγ by immunocompetent mouse cells and human mononuclear cells. Course administration of humic acids to mice enhances the humoral immunity, increasing the number of antibody-forming cells in the spleen and the titer of antibodies in the blood serum after immunization with sheep red blood cells.
Subject(s)
Humic Substances , Soil , Animals , Interleukin-12/metabolism , Interleukin-1beta/metabolism , Macrophages, Peritoneal/metabolism , Mice, Inbred C57BL , Sphagnopsida/chemistry , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Addition of water-soluble polysaccharides isolated from Conium maculatum L. to the mouse peritoneal macrophage culture induces classical activation of antigen-presenting cells due to an increase in NO synthase activity and a decrease in arginase expression.
Subject(s)
Conium/metabolism , Polysaccharides/chemistry , Solubility , Water/chemistry , Alkaloids/metabolism , Animals , Antigen-Presenting Cells , Arginase/metabolism , Female , Immune System , Lymphocytes/metabolism , Mice , Mice, Inbred C57BL , Nitric Oxide/chemistry , PoaceaeABSTRACT
The hemostimulating effects of c-Jun N-terminal kinase (JNK) inhibitor were examined on the mouse model of myelosuppression provoked by 5-fluorouracil. Blockade of JNK during postcytostatic period accelerated recovery of granulomonocytopoiesis and erythropoiesis. It also increased the content of neutrophilic granulocytes and erythroid cells in the hematopoietic tissue and elevated the counts of neutrophils and reticulocytes in the peripheral blood. The development of these phenomena resulted from elevated content and up-regulated functional activity of bone marrow hematopoietic progenitors associated with the direct action of JNK inhibitor on these progenitors and enhanced secretion of hemopoietins by stromal elements of the hematopoiesis-inducing microenvironment.
Subject(s)
Cytostatic Agents/pharmacology , Hematopoietic Stem Cells/drug effects , JNK Mitogen-Activated Protein Kinases/metabolism , Animals , Erythroid Cells/drug effects , Erythroid Cells/metabolism , Fluorouracil/pharmacology , Granulocytes/drug effects , Granulocytes/metabolism , JNK Mitogen-Activated Protein Kinases/antagonists & inhibitors , Mice , Mice, Inbred C57BL , Neutrophils/drug effects , Neutrophils/metabolism , Reticulocytes/drug effects , Reticulocytes/metabolism , Signal Transduction/drug effectsABSTRACT
A course of treatment with humic acids extracted with sodium pyrophosphate from high-moor pine-peat moss-cotton grass peat improves humoral immune response of C57BL/6 mice, stimulates the production of TNFα, IL-1ß, and IL-12 by the animal peritoneal macrophages and the production of IFNγ and TNFα by donor peripheral blood mononuclears, causing no changes in the production of IL-10 in vitro.
Subject(s)
Cytokines/metabolism , Humic Substances , Immunity, Humoral/drug effects , Animals , Cells, Cultured , Humans , Humic Substances/analysis , Immunity, Humoral/physiology , Immunocompetence/drug effects , Leukocytes, Mononuclear/drug effects , Leukocytes, Mononuclear/metabolism , Macrophages, Peritoneal/drug effects , Macrophages, Peritoneal/metabolism , Male , Mice , Mice, Inbred C57BL , Pinus/chemistry , Poaceae/chemistry , Sphagnopsida/chemistryABSTRACT
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
Course treatment with IFN-α2b immobilized on polyethylene glycol stimulates phagocytic activity of peritoneal macrophages and neutrophils, enhances humoral immune response in CBA/CaLac mice, stimulates IL-4 synthesis, and suppresses IFN-γ production by mitogenstimulated splenocytes from experimental animals.
Subject(s)
Adjuvants, Immunologic/administration & dosage , Drug Carriers/administration & dosage , Interferon-alpha/administration & dosage , Macrophages, Peritoneal/drug effects , Polyethylene Glycols/administration & dosage , Animals , Antigen Presentation/drug effects , Cells, Cultured , Drug Evaluation, Preclinical , Immunity, Cellular/drug effects , Immunity, Humoral/drug effects , Interferon alpha-2 , Macrophages, Peritoneal/immunology , Male , Mice, Inbred CBA , Neutrophils/drug effects , Neutrophils/immunology , Phagocytosis/drug effects , Spleen/drug effects , Staphylococcus aureus/immunologyABSTRACT
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 PI3K, MAPK/EPK1/2 and p38-dependent signaling cascade under stimulation of the functions of mesenchymal progenitor cells by fibroblast growth factor. The important role of PI3K and EPK1/2 in the realization of their growth potential has been revealed. Experiments have demonstrated a blocking effect of PI3K inhibitor on proliferation and differentiation of progenitor cells and EPK1/2 inhibitor on mitotic activity of the progenitor cells. The p38-mediated MAPK pathway did not play a role in determining the proliferative and differentiation status of fibroblast CFU.
Subject(s)
Fibroblast Growth Factor 2/pharmacology , Mesenchymal Stem Cells/drug effects , Mitogen-Activated Protein Kinase 1/genetics , Mitogen-Activated Protein Kinase 3/genetics , Phosphatidylinositol 3-Kinases/genetics , p38 Mitogen-Activated Protein Kinases/genetics , Animals , Bone Marrow Cells/cytology , Bone Marrow Cells/drug effects , Bone Marrow Cells/metabolism , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Gene Expression Regulation , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Mice , Mice, Inbred CBA , Mitogen-Activated Protein Kinase 1/antagonists & inhibitors , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/antagonists & inhibitors , Mitogen-Activated Protein Kinase 3/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Phosphoinositide-3 Kinase Inhibitors , Primary Cell Culture , Protein Kinase Inhibitors/pharmacology , Signal Transduction , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
We studied the in vitro role of NF-κB-dependent signaling in the growth capacity of mesenchymal progenitor cells upon stimulation by basic fibroblast growth factor. Proliferative activity of progenitor cells was suppressed by specifi c inhibitors of this transcription factor (NF-κB), oridonin and aurothiomalate. These inhibitors had no effect on differentiation of fibroblast CFU.
Subject(s)
Fibroblast Growth Factor 2/physiology , Mesenchymal Stem Cells/physiology , NF-kappa B/metabolism , Animals , Cell Cycle Checkpoints , Cell Proliferation , Cells, Cultured , Mice, Inbred CBA , Second Messenger SystemsABSTRACT
We studied the role of cAMP- and IKK-2-dependent pathways under conditions of functional stimulation of mesenchymal progenitor cells by basic fibroblast growth factor. An adenylate cyclase inhibitor 2',5'-dideoxyadenosine had no effect on functional activity of early (primitive) mesenchymal precursors. This agent produced a stimulatory effect on proliferation of stromal precursor cells. Mitotic activity of mesenchymal progenitor cells was shown to decrease after treatment with a specific blocker of the IKK-2 kinase-inhibitor complex.
Subject(s)
Cyclic AMP/metabolism , Fibroblast Growth Factor 2/pharmacology , I-kappa B Kinase/metabolism , Mesenchymal Stem Cells/drug effects , Mesenchymal Stem Cells/metabolism , Signal Transduction/physiology , Animals , Cells, Cultured , Mesenchymal Stem Cells/cytology , Mice , Regenerative MedicineABSTRACT
We studied the mechanisms of regenerative (wound healing) effects of songorine associated with functional activation of mesenchymal progenitor cells. The key role of FGF receptors on these progenitor cells in the stimulation of realization of their growth potential under the effect of the alkaloid was demonstrated. Under in vitro conditions, the antibodies to FGF receptor abolished the songorine-induced increase in the number of fibroblast colony-forming units in bone marrow cell culture. The intensity of differentiation of mesenchymal precursors remained unchanged.
Subject(s)
Aconitum/chemistry , Alkaloids/pharmacology , Receptors, Fibroblast Growth Factor/genetics , Skin/drug effects , Wound Healing/drug effects , Wounds, Nonpenetrating/drug therapy , Alkaloids/isolation & purification , Animals , Animals, Outbred Strains , Antibodies/pharmacology , Cells, Cultured , Fibroblast Growth Factors/genetics , Fibroblast Growth Factors/metabolism , Gene Expression , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/drug effects , Mesenchymal Stem Cells/metabolism , Mice , Receptors, Fibroblast Growth Factor/antagonists & inhibitors , Receptors, Fibroblast Growth Factor/metabolism , Signal Transduction , Skin/injuries , Skin/metabolism , Wound Healing/genetics , Wounds, Nonpenetrating/metabolism , Wounds, Nonpenetrating/pathologyABSTRACT
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
We studied the role of intracellular signal molecules PI3K, MAPK ERK1/2, and p38 in the realization of the growth potential of mesenchymal progenitor elements. Under in vitro conditions, PI3K и ERK1/2 specifi c inhibitors reduced fi broblastic colony- and cluster-formation and considerably suppressed proliferative activity of mesenchymal precursors. Blocker of p38 and protein kinase B had no effect on the function of fi broblast CFU.
