ABSTRACT
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.
Subject(s)
Immunoglobulin E , Polysaccharides , Saussurea , Th2 Cells , Saussurea/chemistry , Animals , Th2 Cells/immunology , Th2 Cells/drug effects , Mice , Immunoglobulin E/immunology , Immunoglobulin E/blood , Polysaccharides/pharmacology , Polysaccharides/isolation & purification , Polysaccharides/chemistry , Interleukin-4/immunology , Interleukin-4/metabolism , Immunoglobulin G/immunology , Immunoglobulin G/blood , Plant Extracts/pharmacology , Plant Extracts/chemistry , Anaphylaxis/immunology , Anaphylaxis/drug therapy , Anaphylaxis/chemically induced , Interferon-gamma/immunology , Mice, Inbred BALB C , Mast Cells/drug effects , Mast Cells/immunology , Ovalbumin/immunology , Female , Anti-Allergic Agents/pharmacology , Anti-Allergic Agents/isolation & purificationABSTRACT
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
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.
Subject(s)
Cell Differentiation , Cell Proliferation , Diterpenes , Neural Stem Cells , Receptors, Fibroblast Growth Factor , STAT3 Transcription Factor , Signal Transduction , Neural Stem Cells/drug effects , Neural Stem Cells/metabolism , Neural Stem Cells/cytology , Animals , STAT3 Transcription Factor/metabolism , Receptors, Fibroblast Growth Factor/metabolism , Receptors, Fibroblast Growth Factor/agonists , Signal Transduction/drug effects , Cell Proliferation/drug effects , Diterpenes/pharmacology , Cell Differentiation/drug effects , NF-kappa B/metabolism , Fibroblast Growth Factor 2/metabolism , Fibroblast Growth Factor 2/pharmacology , Tumor Suppressor Protein p53/metabolism , Tumor Suppressor Protein p53/agonists , Phosphatidylinositol 3-Kinases/metabolism , Alkaloids/pharmacology , MAP Kinase Signaling System/drug effects , Janus Kinases/metabolism , Cyclic AMP/metabolism , Cells, Cultured , RatsABSTRACT
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.
Subject(s)
Cyclic AMP-Dependent Protein Kinases , Ethanol , Ethanol/toxicity , Cyclic AMP-Dependent Protein Kinases/genetics , Cyclic AMP-Dependent Protein Kinases/metabolism , Cyclic AMP/metabolism , Signal Transduction , Astrocytes/metabolismABSTRACT
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 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 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 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.
Subject(s)
Ethanol , Janus Kinases , Microglia , STAT3 Transcription Factor , Animals , Astrocytes/drug effects , Astrocytes/metabolism , Astrocytes/pathology , Ethanol/toxicity , Janus Kinases/metabolism , Mice , Microglia/drug effects , Microglia/metabolism , Microglia/pathology , Nerve Growth Factors/genetics , Nerve Growth Factors/metabolism , Neurodegenerative Diseases/genetics , Neurodegenerative Diseases/metabolism , Neurodegenerative Diseases/pathology , Neuroglia/metabolism , STAT3 Transcription Factor/genetics , STAT3 Transcription Factor/metabolismABSTRACT
We studied the role of NF-κB-, cAMP/PKA-, JAKs/STAT3-, ERK1/2-, p38-, JNK- and p53- mediated signaling pathways in the realization of the growth potential of neural stem cells and committed neuronal progenitors under in vitro conditions. The method of pharmacological blockade with selective inhibitors of individual signaling molecules revealed some principal differences in their role in the determination of the proliferation and differentiation status of progenitor cells of different classes. Analysis of the peculiarities of intracellular signaling in cells and comparison of the role of its individual elements attest to the prospects of developing new drugs with neuroregenerative activity based on STAT3 inhibitors or JNK activators. These modulations of activity of signaling molecules can stimulate the realization of the growth potential of committed neuronal progenitors and neutral stem cells, respectively. The blockade of STAT3 and an increase in the content of phosphorylated forms of JNK had no "negative" effects on the functioning of multipotent neural stem cells and committed neuronal progenitors, respectively.
Subject(s)
Neural Stem Cells/metabolism , Neurodegenerative Diseases/metabolism , Regenerative Medicine/methods , Animals , Humans , STAT3 Transcription Factor/metabolism , Signal Transduction/genetics , Signal Transduction/physiologyABSTRACT
We studied the participation of JNK and p53 in the realization of the growth potential of different types of progenitors of the subventricular zone of mouse brain and secretion of neurotrophins by glial cells. The stimulating role of these signaling molecules in mitotic activity and specialization of multipotent neural stem cells was shown. It was found that JNK and p53 do not participate in the regulation of committed neuronal progenitor cells (clonogenic PSA-NCAM+ cells). A dependence of neurotrophic growth factors in individual populations of neuroglia on activity of these protein kinase and transcription factor was revealed. The role of JNK and p53 in astrocytes consists in stimulation of their secretion, and in microglial cells, on the contrary, in its inhibition. The secretory neurotrophic function of oligodendrogliocytes is not associated with JNK and p53 activity.
Subject(s)
Astrocytes/metabolism , MAP Kinase Kinase 4/genetics , Multipotent Stem Cells/metabolism , Neural Stem Cells/metabolism , Neuroglia/metabolism , Tumor Suppressor Protein p53/genetics , Animals , Astrocytes/cytology , Astrocytes/drug effects , Benzothiazoles/pharmacology , CD56 Antigen/genetics , CD56 Antigen/metabolism , Culture Media, Conditioned/pharmacology , Gene Expression Regulation , Lateral Ventricles/cytology , Lateral Ventricles/drug effects , Lateral Ventricles/metabolism , MAP Kinase Kinase 4/metabolism , Mice , Mice, Inbred C57BL , Multipotent Stem Cells/cytology , Multipotent Stem Cells/drug effects , Nerve Growth Factors/biosynthesis , Nerve Growth Factors/genetics , Neural Cell Adhesion Molecules/genetics , Neural Cell Adhesion Molecules/metabolism , Neural Stem Cells/cytology , Neural Stem Cells/drug effects , Neuroglia/cytology , Neuroglia/drug effects , Signal Transduction , Toluene/analogs & derivatives , Toluene/pharmacology , Tumor Suppressor Protein p53/metabolismABSTRACT
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
Under conditions of steady-state hemopoiesis, nuclear factor NF-κB, in contrast to MAP kinase p38, plays an important role in the maintenance of the initial level of secretory activity of monocytes. The increase in the production of G-CSF under stress conditions (10-h immobilization) is mainly regulated by the alternative p38MARK signaling pathway via activation of p38 synthesis. It was shown that under conditions of cytostatic-induced myelosuppression, the production of protein kinase p38 in cells decreases, and it, like NF-κB, is not the main one in the production of hemopoietin by mononuclear phagocytes.
Subject(s)
Cell Differentiation , Intracellular Signaling Peptides and Proteins/physiology , Phagocytes/physiology , Animals , Bone Marrow Cells/physiology , Granulocyte Colony-Stimulating Factor/metabolism , Intracellular Signaling Peptides and Proteins/metabolism , MAP Kinase Signaling System/physiology , Male , Mice , Mice, Inbred C57BL , Mitogen-Activated Protein Kinases/physiology , NF-kappa B/metabolism , Phagocytes/metabolism , Signal Transduction/physiology , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
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
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
The role of JAK/STAT3-mediated signaling pathway in the realization of the growth potential of mesenchymal precursor cells was examined in vitro. The stimulating role of JAKs and STAT3 towards proliferating activity of progenitor cells and their different role in the regulation of differentiation of the progenitor elements were demonstrated. Inhibitors of JAKs and STAT3 reduced the yield of fibroblast CFU and their mitotic activity. Blockade of JAKs accelerated and selective inactivation of STAT3 decelerated differentiation of progenitor cells.
Subject(s)
Bone Marrow Cells/metabolism , Fibroblasts/metabolism , Janus Kinase 1/genetics , Janus Kinase 3/genetics , Mesenchymal Stem Cells/metabolism , STAT3 Transcription Factor/genetics , Animals , Anthraquinones/pharmacology , Bone Marrow Cells/cytology , Bone Marrow Cells/drug effects , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Fibroblasts/cytology , Fibroblasts/drug effects , Gene Expression Regulation , Janus Kinase 1/antagonists & inhibitors , Janus Kinase 1/metabolism , Janus Kinase 3/antagonists & inhibitors , Janus Kinase 3/metabolism , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/drug effects , Mice , Mice, Inbred C57BL , Nitriles , Phosphorylation , Primary Cell Culture , Pyrazoles/pharmacology , Pyrimidines , STAT3 Transcription Factor/antagonists & inhibitors , STAT3 Transcription Factor/metabolism , Signal Transduction , Sulfonamides/pharmacologyABSTRACT
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.