ABSTRACT
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(-)).
Subject(s)
Enzymes, Immobilized/administration & dosage , Hyaluronoglucosaminidase/administration & dosage , Pulmonary Fibrosis/pathology , Stem Cells/metabolism , Animals , Antigens, CD/metabolism , Bone Marrow/immunology , Bone Marrow/pathology , Lung/pathology , Mice, Inbred C57BL , Pulmonary Fibrosis/drug therapy , Pulmonary Fibrosis/immunology , Stem Cells/drug effectsABSTRACT
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).
Subject(s)
Dopamine Antagonists/pharmacology , Fibroblasts/drug effects , Mesenchymal Stem Cells/drug effects , Pulmonary Alveoli/drug effects , Pulmonary Fibrosis/drug therapy , Spiperone/pharmacology , Animals , Antigens, CD/metabolism , Biomarkers/metabolism , Bleomycin , Bone Marrow Cells/cytology , Bone Marrow Cells/drug effects , Bone Marrow Cells/metabolism , Cell Lineage/drug effects , Endothelial Cells/cytology , Endothelial Cells/drug effects , Endothelial Cells/metabolism , Fibroblasts/metabolism , Fibroblasts/pathology , Gene Expression , Granulocytes/cytology , Granulocytes/drug effects , Granulocytes/metabolism , Hematopoietic Stem Cells/cytology , Hematopoietic Stem Cells/drug effects , Hematopoietic Stem Cells/metabolism , Macrophages/cytology , Macrophages/drug effects , Macrophages/metabolism , Megakaryocytes/cytology , Megakaryocytes/drug effects , Megakaryocytes/metabolism , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Mice , Mice, Inbred C57BL , Neutrophil Infiltration/drug effects , Pulmonary Alveoli/metabolism , Pulmonary Alveoli/pathology , Pulmonary Fibrosis/chemically induced , Pulmonary Fibrosis/metabolism , Pulmonary Fibrosis/pathologyABSTRACT
We studied differentiation of multipotent mesenchymal stromal cells (MMSC) of the lungs of C57Bl/6 mice with bleomycin-induced pneumofibrosis. Adherent mononuclear cells found in mouse lungs demonstrated mesenchymal phenotype and expressed CD44, CD73, CD90, and CD106, but not CD31, CD34, and CD45. The cells with MMSC characteristics differentiate in vitro into various cells of stromal lines (chondrocytes, osteogenic cells, adipocytes, and fibroblasts). Bleomycin increased the growth rate of MMSC and selectively promoted their differentiation towards fibroblast cells.
Subject(s)
Cell Differentiation/physiology , Fibrosis/pathology , Lung Diseases/pathology , Lung/cytology , Mesenchymal Stem Cells/cytology , Adipocytes/cytology , Animals , Chondrocytes/cytology , Fibroblasts/cytology , Leukocytes, Mononuclear/cytology , Mice , Mice, Inbred C57BLABSTRACT
Antifibrotic activity of testicular hyaluronidase, immobilized on polyethylenoxide and obtained by electron beam synthesis, was studied on the model of bleomycin injuries to the alveolar epithelium (irreversible pneumofibrosis) in C57Bl/6 mice and compared to the effect of testicular hyaluronidase. Intranasal therapy with immobilized and testicular hyaluronidases prevented the deposition of fibrotic mass in the parenchyma of "bleomycin" lungs. The effect of immobilized hyaluronidase was more pronounced than that of testicular hyaluronidase. The studied compounds were virtually inessential for infiltration of the alveolar interstitium and alveolar tracts by lymphocytes, macrophages, neutrophils, and plasma cells. Unchanged histoarchitectonics of bleomycin-damaged lungs in immobilized hyaluronidase therapy was due to suppression of the progenitor fibroblast cells (CD45(-)).
Subject(s)
Anti-Inflammatory Agents/administration & dosage , Hyaluronoglucosaminidase/administration & dosage , Idiopathic Pulmonary Fibrosis/drug therapy , Animals , Anti-Inflammatory Agents/chemistry , Bleomycin , Drug Carriers/administration & dosage , Drug Carriers/chemistry , Enzymes, Immobilized/administration & dosage , Enzymes, Immobilized/chemistry , Hyaluronoglucosaminidase/chemistry , Idiopathic Pulmonary Fibrosis/chemically induced , Idiopathic Pulmonary Fibrosis/pathology , Mice , Mice, Inbred C57BL , Polyethylenes/chemistry , Pulmonary Alveoli/pathology , Respiratory Mucosa/pathologyABSTRACT
The possibility of boosting antifibrotic activity of testicular hyaluronidase immobilized on polyethylene oxide with spiperone was studied on the bleomycin models of a single (partially reversible pneumofibrosis) and repeated (irreversible pneumofibrosis) injuries to the alveolar epithelium in C57Bl/6 mice. The antifibrotic effect was more pronounced after successive treatment with immobilized hyaluronidase and spiperone than after individual treatment with each of the compounds: no collagen deposition in the parenchyma of bleomycin-damaged lungs was found. The decrease in inflammatory cell (lymphocytes, macrophages, neutrophils, plasma cells) infiltration of the alveoli and alveolar tracts interstitium in mice treated by immobilized hyaluronidase and spiperone did not differ from the anti-inflammatory effect of spiperone monotherapy.