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We studied the age-related characteristics of the response of stem cells and liver in male Wistar rats to administration of carbon tetrachloride (CCl4) and ethanol. It was shown that modeling of liver cirrhosis caused inflammation, fibrosis, damage to sinusoidal capillaries, necrosis, and disturbances in the functional activity of hepatocytes in young rats. These processes were accompanied by mobilization of profibrotic mesenchymal stem cells (MSC), proinflammatory hematopoietic stem cells (HSC) and lymphocytes (CD45hiCD133+) from the bone marrow into the blood and migration to the liver. On the other hand, the number of hepatocyte precursors expressing Sox9 (cells of Hering's canal), immature cholangiocytes, Ito cells, oval cells, and endothelial cells of the liver sinusoids) sharply increased in the liver. In young rats, mobilization and migration of MSC, HSC, and hepatocyte precursors against the background of liver cirrhosis were more intensive than in old animals. The higher resistance of old rats to exposure is associated with age-related changes in the niches as well as in mobilization and migration of cells.

The model of streptozotocin-induced diabetes mellitus in C57Bl/6 mice was employed to study the role of precursors of insulin-producing β-cells, hematopoietic stem cells, and progenitor hematopoietic cells in inflammation. In addition to provoking hyperglycemia, streptozotocin elevated serum levels of IL-1β and hyaluronic acid, induced edema in the pancreatic insular tissue and its infiltration by inflammatory cells (neutrophils, lymphocytes, and macrophages) and fibroblasts. Inflammation in pancreatic islets was accompanied by necrotic processes and decreasing counts of multipotent progenitor β-cells (CD45(-), TER119(-), c-kit-1(-), and Flk-1(-)), oligopotent progenitor β-cells (CD45(-), TER119(-), CD133(+), and CD49f(low)), and insulinproducing β-cells (Pdx1(+)). Pancreatic infl ammation was preceded by elevation of the number of short-term hematopoietic stem cells (Lin-Sca-1(+)c-kit(+)CD34(+)) relative to long-term cells (Lin(-)Sca-1(+)c-kit(+)CD34(-)) in the bone marrow as well as recruitment of hematopoietic stem and progenitor cells into circulation. Transplantation of bone marrow hematopoietic stem and progenitor cells from diabetic C57Bl/6 donor mice to recipient CBA mice with 5-fluorouracilinduced leukopenia accelerated regeneration of granulocytopoiesis in recipient mice.

The influence of low-molecular-weight sodium alginate, which is administered as an isolated agent and in combination with cyclophosphamide, on the parameters of peripheral blood and bone marrow was studied in mice with Lewis lung carcinoma. It was shown that administration of sodium alginate with a molecular weight of 1–10 and 20–30 kDa to tumor-bearing animals prevents bone marrow failure by activating the process of regeneration of granulocytic hemopoietic stem cells that are damaged by a single injection or repetitive injections of a cytostatic agent, due to stimulation of the clonal activity of granulocytopoiesis precursors. As a result, this treatment prevents the progression of leukopenia.

The antifibrotic properties of spiperone and its effect on stem and progenitor cells were studied on the model of reversible bleomycin-induced pulmonary fibrosis in C57Bl/6 mice. Spiperone reduced infiltration of the alveolar interstitium and alveolar ducts with inflammatory cells and prevented the growth of the connective tissue in the parenchyma of bleomycin lungs. Apart from anti-inflammatory effect, spiperone suppressed bone marrow hemopoietic cells (CD3, CD45R (B220), Ly6C, Ly6G (Gr1), CD11b (Mac1), TER-119)-, Sca-1+, c-Kit+, CD34- and progenitor hemopoietic cells (granulocyte-erythroid-macrophage-megakaryocytic and granulocyte CFU). Spiperone-induced disturbances of fi brogenesis were paralleled by restoration of endothelial cells in the lung parenchyma, reduction of the number of circulating bone marrow cells and lung mesenchymopoietic cells (mesenchymal multipotent stromal cells (CD31-, CD34-, CD45-, CD44+, CD73+, CD90+, CD106+) and progenitor fi broblast cells), and suppression of multilineage differentiation of multipotent mesenchymal stromal cells (including fi broblast-lineage cells).

Effects of a sympatholytic drug on bone marrow stromal and hemopoietic precursors were studied on the model of cyclophosphamide-induced myelosuppression. Sympatholytic treatment increased the content of hemopoietic stem cells of different classes in the bone marrow. Selective stimulation of differentiation of polypotent precursors into granulocyte-macrophage precursors was noted. Acceleration of proliferation and maturation of granulocytic precursors was observed at later terms during regeneration of the hemopoietic tissue. The sympatholytic inhibited proliferation of stromal precursors and reduced feeder activity of fibroblasts for granulocyte precursors.

On the model of toxic diffuse pulmonary fibrosis induced by intratracheal administration of bleomycin, we studied reactions of the blood system, content of stem cells, committed hemopoietic and stromal progenitor cells in the bone marrow, spleen and peripheral blood of C57Bl/6 mice. It was shown that the development of diffuse pulmonary fibrosis was accompanied by hyperplasia of bone marrow hemopoiesis and leukocytosis in the peripheral blood. Activation of the erythroid and granulocytic hemopoietic stems was related to stimulation of hemopoietic stem cells (polypotent cells, granulocyte/erythroid/macrophage/megakaryocyte precursor cells) and committed erythroid and myeloid progenitor cells in the bone marrow. At the same time, the number of stromal precursors increased. Bleomycin increased the count of hemopoietic stem cells the peripheral blood and spleen and reduced the content of mesenchymal stem cells in the spleen and bone marrow.

Effects of serotonin and adrenomimetics (phenylephrine and isoprenaline) on bone marrow stromal and polypotent hemopoietic precursors were studied in vitro on the model of cyclophosphamide- induced myelosuppression. It was found that under conditions of myelosuppression, adrenomimetics potentiate differentiation of polypotent hemopoietic precursors into mature precursors (granulocyte-macrophage and granulocyte CFU) initiated by granulocytic CSF, while serotonin suppresses these processes. Adrenomimetics (especially, isoprenaline) abolish high rate of division of stromal precursors and suppress the growth of granulocytic CSF induced by fibroblast-like cells. Serotonin does not affect proliferation of stromal precursors, but potentiates the granulocytopoiesis-stimulating effects of fibroblasts.

Intraperitoneal injection of paclitaxel (Mitotax) in a single dose of 40 mg/kg was followed by an increase in the number of mitotic granulocytic and erythroid cells, hypoplasia, and pancytopenia of the bone marrow in CBA/CaLac mice. The test preparation decreased the number of hematopoietic precursor cells for erythropoiesis and granulocytopoiesis, but increased the count of polyploid cells and incidence of structural and genomic abnormalities in bone marrow cells. These changes were reversible.

We studied monoaminergic mechanisms of regulation of the pool of erythropoietic precursors in active and passive mice under conditions of conflict situation and during paradoxical sleep deprivation. It was found that in animals with different behavior subjected to experimental neuroses the regulatory effects of CNS monoamines on proliferation and differentiation are mediated via adrenergic and erythropoietin-sensitive structures on erythroid stem cells. The revealed intergroup differences in precursor proliferation and differentiation rates (and erythrokaryocyte content in the bone marrow) are related to peculiarities in the activity of monoaminergic system and their interaction with peripheral structures on erythropoietic precursors.

We studied the local mechanisms of neurosis-associated changes in the granulocytic hemopoietic stem in CBA/CaLac mice with different learning capacity. It was found that in good learners hyperplasia of the bone marrow granulocytopoiesis during neurosis was related to enhanced proliferative activity and accelerated maturation of granulocyte-macrophage precursors resulting from increased formation of granulocytic and erythrogranulocytic hemopoietic islets. In poor learners primary suppression of the bone marrow granulocytopoiesis was associated with impaired formation of hemopoietic islets (conflict situation) and inhibition of differentiation of granulocyte-macrophage precursors (paradoxical sleep deprivation followed by T-maze learning). Then, recovered ability of bone marrow cells to form granulocyte complexes even against the background of inhibition of precursor differentiation (conflict situation) and accelerated maturation of granulocytic cells (paradoxical sleep deprivation and maze learning) lead to hyperplasia of the granulocytic hemopoietic stem.

We studied changes in the erythroid hemopoietic stem in CBA/CaLac mice with experimental neuroses demonstrating good and poor learning capacities (conflict situation and paradoxical sleep deprivation followed by training in a 3-arm T-maze). The animals with different learning capacities exhibited pronounced hyperplasia of the erythroid hemopoietic stem in response to neurosis. Activation of the erythron in good learners was related to acceleration of division and maturation of erythroid precursors and enhanced formation of cell complexes containing central macrophage. In poor learners hyperplasia of the erythroid hemopoietic stem under conditions of conflict situation manifested in activation of proliferation and differentiation (against the background of decreased count of erythroid and mixed complexes in the bone marrow), while after paradoxical sleep deprivation followed by T-maze training this hyperplasia was associated with increased formation of additional hemopoietic islets (against the background of desynchronization of division and maturation of erythroid precursor cells).

We studied the effects of adaptogens extracts of Siberian ginseng, Rhodiola rosea, bergenia, and ginseng and pantohematogen, on erythropoiesis after paradoxical sleep deprivation. Adaptogens stimulated bone marrow erythropoiesis in the early stage, but decreased the count of bone marrow erythrokaryocytes 3-7 days after treatment. The effect of adaptogens on erythropoiesis is associated with modulation of the state of brain neurotransmitter systems followed by changes in functional activity of cells in the hemopoiesis-inducing microenvironment.

Bleomycin-induced lung inflammation and fibrosis were assessed in a model of idiopathic pulmonary fibrosis in C57BL/6 mice and effect of spiperone on histopathological lung indicators, hematopoietic (HSC) and mesenchymal (MSC) stem cells and progenitor cells was characterized. Spiperone is a selective antagonist of D2 dopamine receptors which disturbs dopamine mediation. Hematoxylin and eosin staining showed spiperone decreased alveolar epithelial edema, exudation and infiltration of the alveoli walls and lumen by inflammatory cells (neutrophils, macrophages, plasma cells) after bleomycin instillation. Picrofuchsin staining by Van Gieson revealed spiperone decreases the area of connective tissue in the lung fibrotic phase of pulmonary fibrosis. ELISA assay determined the decrease levels of collagen type I, hydroxyproline and total collagen in lung homogenate after spiperone treatment. Number of “long-term” HSCs (Lin- Sca-1+c-Kit+CD34-), «short-term” HSCs (Lin-Sca-1+c-Kit+CD34+), hematopoietic progenitor cells and MSC-like cells in lung with pneumofibrosis decreased after spiperone treatment. This spiperone effect we connect to the violation of immature bone marrow cells migration. Additionally, spiperone inhibited clonal activity of hematopoietic (CFU-GEMM, CFU-G) and mesenchymal (CFU-F) progenitor of bone marrow, blood and lungs. An additional feature of the spiperone action was the ability to decrease the capacity of self-renewal and MSCs differentiation activity into adipocytes, osteoblasts, chondrocytes and fibroblast cells. Thus, a selective antagonist D2 of dopamine receptors spiperone can act as a potential antifibrotic agent for the treatment of toxic pulmonary fibrosis. The overall conclusion was that the neurotropic agent spiperone is able to influence the stem and progenitor cells in lung pathology effectively.

The effect of course treatment with neuroleptic haloperidol on the inflammatory response and state of the connective tissue in the lungs of C57Bl/6 mice was studied on the model of toxic pulmonary fibrosis induced by intratracheal administration of bleomycin. This neuroleptic decreased the inflammatory response and reduced the growth of the connective tissue in the lungs. The anti-inflammatory effect of haloperidol is related to a decrease in activity of bone marrow hemopoietic stem cells and committed hemopoietic precursors. The antifibrotic effect of this drug is associated with inhibition of mesenchymal precursor cells.

Hemopoiesis-stimulating activity of immobilized oligonucleotide preparation was studied on the model of cytostatic myelosuppression induced by injection of cyclophosphamide and 5-fluorouracil. Immobilized oligonucleotides stimulated regeneration of erythro- and granulocytopoiesis in the bone marrow under conditions of cytostatic treatment. The counts of neutrophilic granulocytes and platelets in the peripheral blood increased. The stimulatory effect of the drug was more manifest in animals with active behavior. The mechanism of immobilized oligonucleotide effect was based on stimulation of functional activity of erythroid and granulocytic macrophage precursors.

The influence of granulocyte colony-stimulating factor (G-CSF) has been studied on a model of bleomycin-induced pulmonary fibrosis. It is established that G-CSF significantly increases infiltration of alveolar and alveolar duct interstitium by inflammation cells (lymphocytes, neutrophils, plasmocytes) and increases collagen deposition in lung under conditions of bleomycin introduction. Simultaneously with profibrotic and anti-inflammation effects, G-CSF increased the content of granulocyte cells in the bone marrow and peripheral blood, which was related to the stimulation of committed granulocyte precursors in the bone marrow.

The effect of immobilized hyaluronidase on stem and progenitor cells of the lungs was studied on the model of partially reversible toxic bleomycin-induced pulmonary fibrosis in C57Bl/6 mice. During the inflammation phase, immobilized hyaluronidase reduced infiltration of alveolar interstitium with hemopoietic stem cells Sca-1(+), c-Kit(+), CD34(-), (CD3, CD45R (B220), Ly6C, Ly6G (Gr1), CD11b (Mac1), TER-119)(-). Improvement of histological parameters of bleomycin lungs during the phase of collagen fiber deposition after the treatment was accompanied by accumulation of mesenchymal multipotent stromal cells (CD31(-), CD34(-), CD45(-), CD44(+), CD73(+), CD90(+), CD106(+)decrease in the population of pan-hemopoietic cells (CD45(+)), accelerated restoration of the content of endothelial cells, and inhibition of clonal activity of fibroblast precursors (CD45(-)).

Experiments were performed on the model of cytostatic myelosuppression induced by cyclophosphamide. We compared the effect of immobilized granulocyte CSF (the preparation was created in Russia) and reference standard preparation of granulocyte CSF on the development of neutrophilic leukopenia and hemopoietic precursors of various classes. It was found that preparations of granulocyte CSF decreased the duration and degree of peripheral blood neutropenia. The granulocytopoiesis-stimulating effect was related to stimulation of multipotent hemopoietic precursors, granulocyte-erythroid-macrophage-megakaryocyte precursors, and granulocyte precursors. Induction of division and maturation of multipotent hemopoietic precursors, granulocyte-erythroid-macrophage-megakaryocyte precursors, and granulocyte precursors and recovery of cellularity of the granulocytic hemopoietic stem after administration of immobilized granulocyte CSF were observed at later terms compared to treatment with the reference preparation of granulocyte CSF.

Effect of the antiserotonin drug cyproheptadine on the erythropoiesis has been studied under conditions of the administration of cytostatics (fluoropyrimidine antimetabolite 5-fluorouracil (5-FU), alkylating agent cyclophosphane) with different mechanisms of action. It is established that the antiserotonin drug significantly accelerates regeneration of the erythroid hemopoietic branch, especially in the case of 5-FU. The depression of erythron under the conditions of cyclophosphane injections was retained. The erythropoiesis-stimulating effect of cyproheptadine is based on the restoration of the structural and functional organization of the bone marrow (formation of erythroid hemopoietic islets).

The influence of ginseng and skullcap (Scutellaria baicalensis) extracts on the local mechanisms of erythropoiesis control under the conditions paradoxical sleep deprivation has been studied. The regulatory action of both preparations on the bone marrow erythropoiesis is based on their ability to influence the proliferation and differentiation in erythroid cells-precursors via modulation of the functional activity of cellular hemopoiesis-inducing microenvironment elements. In particular, the drugs induce changes in the structural and functional organization of the bone marrow and in the production of short-range humoral erythropoiesis regulatory elements.

We studied the effect of bone marrow Thy 1,2+ cells on the growth of erythroid colonies and the role of monoamine in the regulation of proliferation and differentiation of erythroid precursor cells in experimental neuroses. Under conditions of conflict situation Thy 1,2+ cells stimulated the growth of erythropoietic precursors (directly and via interaction with adherent cells of the hemopoiesis-inducing environment). Under conditions of paradoxical sleep deprivation the absence of interaction between Thy 1,2+ cells and adherent bone marrow fraction disturbs erythroid colony formation. In experimental neuroses the state of erythropoietic precursors and secretory activity of elements of the hemopoiesis-inducing microenvironment are controlled by monoamines. The regulatory effect of monoamines is realized via adrenergic and erythropoietin-sensitive receptors on erythroid precursors and cells of the hemopoiesis-inducing microenvironment.

We studied the direct and stromal cell-mediated effects of bone marrow Thy 1,2+ cells on the growth of granulocyte-macrophage and erythroid colonies from the bone marrow of CBA/CaLac mice with experimental neuroses (conflict situation and paradoxical sleep deprivation). Proliferation and differentiation of hemopoietic precursors during neuroses are controlled by regulatory T cells. In conflict situation Thy 1,2+ cells stimulate the growth of hemopoietic precursors, which is associated with their direct effect and interaction with adherent cells of the hemopoiesis-inducing microenvironment. The interaction of Thy 1,2+ cells with adherent bone marrow cells during paradoxical sleep deprivation stimulates the formation of only granulocyte-macrophage colonies.

In this study was found that inflammation in interstitial lung tissue of mice C57Bl/6 after intratracheal injection of the bleomycin is accompanied by the increase of the bone marrow and circulating blood cells with the phenotype (CD3, CD45R (B220), Ly6C, Ly6G (Gr1), CD11b (Mac1), TER-119), Sca-1 + , c-Kit + , CD34 - (hematopoietic stem cells (HSCs)) and hematopoietic progenitor cells (granulocyte-erythroid-macrophage-megakaryocytic (CFU-GEMM) and granulocyte precursors (CFU-G)). These results demonstrate that the intraperitoneal reserpine injection reduces alveolar interstitium and alveolar passages infiltration by inflammatory cells and prevents the connective tissue growth in the lung parenchyma. Supposedly, the reserpine anti-inflammatory effect is caused by the reduced activity of the bone marrow HSCs differentiation in CFU-G, the decrease of circulating HSCs and progenitor hematopoietic cells and the violation of their migration in bleomycin-treated lungs. These data suggest the decrease caused by the reserpine in deposition of collagen fibers in the lung's parenchyma associated with the decrease in the inflow of multipotent mesenchymal stromal cells (MSCs) and fibroblast progenitor bone marrow-derived cells to lungs. Thus, reserpine violates the MSCs differentiation into fibroblast-like cells.

We studied the role of the serotoninergic system in the regulation of the granulocytic hemopoietic stem during cytostatic myelosuppressions caused by administration of cyclophosphamide or 5-fluorouracil. After cytostatic treatment, the state of granulocytopoietic precursors and functional activity of cells in the hemopoiesis-inducing microenvironment were regulated by serotoninergic structures of the brain. The regulatory influence of CNS was realized via serotonin receptors on precursors and microenvironment cells. The response of the granulocytic stem was determined by specific interaction between distant and local regulatory structures and inhibitory effect of cyclophosphamide and 5-fluorouracil on granulocytic cells and regulatory systems.

The role of central adrenergic structures in the regulation of the erythroid hematopoietic stem was studied during administration of cyclophosphamide and 5-fluorouracil. The central nervous system contributed to suppression of erythropoiesis during cytostatic treatment. The suppressive effect of brain adrenergic structures on the erythron after treatment with cyclophosphamide and 5-fluorouracil was related to dysfunction of adherent cells in the hemopoiesis-inducing microenvironment (formation of hemopoietic islets and secretion of erythropoietic activity) and production of growth factors by myelokaryocytes, respectively. Brain norepinephrine had an inhibitory effect on proliferative activity and differentiation of erythroid precursors that were associated with the erythropoietin and peripheral alpha-adrenergic structures. However, stimulation of beta-adrenergic structures was followed by an increase in the rate of erythroid cell maturation.

Exhaustion of catecholamine depot in the CNS before modeling of cytostatic myelosuppression potentiates the stimulatory effect of granulocytic CSF on regeneration of the granulocytic hemopoiesis stem. The increase in granulocyte count in the blood system under these conditions is caused by recovery of the structural and functional organization of the hemopoietic tissue (at the expense of reduction of the suppressive effect of catecholamines on the formation of granulocytic hemopoietic islets) and simultaneous stimulation of division and maturation of granulomonocytic precursors.

The hemopoiesis-stimulating effect of combined treatment with immobilized oligonucleotides and hyaluronidase preparations was studied during cytostatic-induced myelosuppression caused by cyclophosphamide administration. Immobilized hyaluronidase was shown to increase the effi ciency of correction of changes in the erythroid and granulocytic hemopoietic stems with immobilized oligonucleotides. This potentiation of the effect of immobilized oligonucleotides by immobilized hyaluronidase was related to an increase in functional activity of committed hemopoietic precursors.

Combined stimulatory effect of antiserotonin drug and erythropoietin on the bone marrow erythropoiesis was studied under conditions of cytostatic myelosuppression. The stimulatory effect of erythropoietin on regeneration of the hemopoietic erythroid stem increased, if serotonin mediation in the CNS was disordered before induction of cytostatic myelosuppression. The stimulatory effect of a combination of cyproheptadine and epocrine (experimental group) significantly surpassed that of cyproheptadine and epocrine monotherapies. The increase in the content of erythroid elements in the blood system of experimental animals was due to recovery of the structural and functional integrity of the hemopoietic tissue (at the expense of reduction of the suppressive effect of CNS serotonin on the formation of erythroid hemopoietic islets) and simultaneous stimulation of division and maturation of erythroid precursors with epocrine.

The preparation containing ultralow doses of antibodies to stem cell factor considerably activates bone marrow myelopoiesis suppressed by cyclophosphamide. This effect of the preparation is based on stimulation of proliferation of committed hemopoietic precursors and increase in functional activity of adherent elements of the hemopoiesis-inducing microenvironment.

Cells of the erythrocytic series develop hyperplasia in mice exposed to immobilization stress, conflict situation, or conflict situation while being deprived of REM sleep. These mice contain increased numbers of morphologically recognizable erythroid cells and erythroid cell precursors in the bone marrow, and culture media conditioned by their nonadherent or adherent karyocytes shows elevated erythropoietic activity. In contrast, bone marrow erythropoiesis is depressed for a long time in mice deprived of REM sleep and mice with and without experiencing a conflict situation. Probably, enhancement of erythropoiesis is due to activation of adrenergic mechanisms regulating hematopoiesis, while its depression is due to impaired operation of the central adrenergic mechanisms.

Immobilization stress and experimental neurosis caused by conflict situation, deprivation of REM sleep, or their combination caused hyperplasia of the granulocytic stem in the bone marrow. Pronounced changes in hemopoietic tissue resulted from increased secretion of hemopoesin-producing cells of the microenvironment.

The effects of antiserotonin preparation on the development of the connective tissue in the lungs, reaction of the blood system, and the content of hemopoietic stem cells, committed hemopoietic and stromal precursors in BM, spleen, and peripheral blood were studied on C57Bl/6 mice with experimental toxic lung fibrosis caused by intratracheal administration of bleomycin. It was demonstrated that the antiserotonin drug inhibits the growth of the connective tissue in the lungs and attenuates the course inflammatory process primarily due to inhibition of the granulocytic lineage, which was related to suppression of hemopoietic stem cells. Reduced content of the stromal precursor cells in BM and spleen was noted.