ABSTRACT
BACKGROUND: Metastatic disease is a major and difficult-to-treat complication of lung cancer. Considering insufficient effectiveness of existing therapies and taking into account the current problem of lung cancer chemoresistance, it is necessary to continue the development of new treatments. METHODS: Previously, we have demonstrated the antitumor effects of reprogrammed CD8+ T-cells (rCD8+ T-cells) from the spleen in mice with orthotopic lung carcinoma. Reprogramming was conducted by inhibiting the MAPK/ERK signalling pathway through MEKi and the immune checkpoint PD-1/PD-L1. Concurrently, CD8+ T-cells were trained in Lewis lung carcinoma (LLC) cells. We suggested that rCD8+ T-cells isolated from the spleen might impede the development of metastatic disease. RESULTS: The present study has indicated that the reprogramming procedure enhances the survival and cytotoxicity of splenic CD8+ T-cells in LLC culture. In an LLC model of spontaneous metastasis, splenic rCD8 + T-cell therapy augmented the numbers of CD8+ T-cells and CD4+ T-cells in the lungs of mice. These changes can account for the partial reduction of tumors in the lungs and the mitigation of metastatic activity. CONCLUSIONS: Our proposed reprogramming method enhances the antitumor activity of CD8+ T-cells isolated from the spleen and could be valuable in formulating an approach to treating metastatic disease in patients with lung cancer.
Subject(s)
CD8-Positive T-Lymphocytes , Carcinoma, Lewis Lung , Spleen , Animals , Carcinoma, Lewis Lung/immunology , Carcinoma, Lewis Lung/pathology , CD8-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/metabolism , Mice , Spleen/pathology , Spleen/immunology , Lung Neoplasms/immunology , Lung Neoplasms/pathology , Lung Neoplasms/secondary , Mice, Inbred C57BL , Cellular Reprogramming , Cell Line, Tumor , Disease Models, AnimalABSTRACT
The responses of tumor stem cells and various populations of CD4 and CD8 T cells of young and aged C57BL/6 mice were studied in a lung cancer model. Using Lewis lung carcinoma cell line, an orthotopic model of lung cancer was modeled. Cancer stem cells, circulating tumor cells, and various populations of CD4 and CD8 T cells in the blood and lung tissue were studied by cytometry. We revealed age-related differences in the content of various populations of CD4 and CD8 T cells in the blood and lungs of intact young and aged mice. Age-related features of the reaction of various populations of cancer stem cells and CD4 and CD8 T cells in the blood and lungs of animals in the Lewis lung carcinoma were shown.
Subject(s)
Carcinoma, Lewis Lung , Lung Neoplasms , Animals , Mice , Carcinoma, Lewis Lung/pathology , Mice, Inbred C57BL , Lung Neoplasms/pathology , CD8-Positive T-Lymphocytes/metabolism , Neoplastic Stem Cells/metabolismABSTRACT
We studied the effects of the extract of the terrestrial part of Aconitum baicalense in BALB/c female mice at the early stages after the injection of N-methyl-N-nitrosourea (MNU). The extract reduced inflammatory activity and tumor growth in the mammary gland. The antitumor and anti-inflammatory effects of the extract are based on the inhibition of cancer stem cells, hematopoietic stem cells, and hematopoietic progenitor cells that promote inflammation. The extract of A. baicalense disrupted the recruitment of epithelial progenitor cells and angiogenesis precursors to the mammary gland preventing neovascularization and transformation of epithelial cells into tumor cells.
Subject(s)
Aconitum , Adult Stem Cells , Mammary Neoplasms, Experimental , Female , Mice , Animals , Methylnitrosourea , Plant Extracts/pharmacology , Plant Extracts/therapeutic use , Adult Stem Cells/pathology , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/therapeutic use , Mammary Neoplasms, Experimental/chemically induced , Mammary Neoplasms, Experimental/drug therapy , Mammary Neoplasms, Experimental/pathologyABSTRACT
Regenerative processes in the liver were studied in 3-month-old (young) and 9-month-old (aged) male Wistar rats on day 1 after 30 and 70% hepatectomy. Regardless of the resected liver volume, shifts in the biochemical parameters of the serum in aged rats were more pronounced than in young animals. After 30% hepatectomy, no age differences in the rate of hepatic regeneration were found, while after 70% liver resection this parameter was higher in young rats. Hepatectomy in young rats led to recruitment of MSC, hepatocyte precursors, endothelial and epithelial progenitor cells into the liver parenchyma and increased fluidity of the plasma and mitochondrial membranes of hepatocytes. In aged rats, the recruitment of MSC, hepatocyte precursors, and endothelial progenitor cells into the injured liver was impaired and the rigidity of the mitochondrial membranes of hepatocytes increased.
Subject(s)
Hepatectomy , Liver Regeneration , Rats , Male , Animals , Rats, Wistar , Liver/surgery , HepatocytesABSTRACT
Various stem cells were studied in female BALB/c mice at the early terms after administration of N-methyl-N-nitrosourea to search early diagnostic markers and therapeutic targets. At these terms, damage to the epithelium and endothelium, inflammation, and fibrosis were observed in the mammary gland, but the tumor was not detected. Cancer stem cells, hematopoietic stem cells (HSC), hematopoietic progenitor cells, angiogenic precursors, and epithelial progenitor cells were found in the blood and mammary gland. Cancer stem cells (CD44+CD24-) are proposed as the early diagnostic marker of breast cancer, and short-living HSC, hematopoietic progenitor cells, and angiogenic precursors (CD45-CD117+FLK-1+) as predictors of the formation of tumor microenvironment.
Subject(s)
Breast Neoplasms , CD24 Antigen , Animals , Biomarkers, Tumor , Breast Neoplasms/diagnosis , Breast Neoplasms/drug therapy , Breast Neoplasms/pathology , Cell Line, Tumor , Epithelium/pathology , Female , Hematopoietic Stem Cells/pathology , Humans , Hyaluronan Receptors , Mice , Neoplastic Stem Cells/pathology , Tumor MicroenvironmentABSTRACT
The viscosity of plasma and mitochondrial membranes of hepatocytes was studied in young (3-month-old) and old (9-month-old) male Wistar rats. It was shown that viscosity of hepatocyte plasma and mitochondrial membranes in young rats under optimal vital functions in the area of protein-lipid membrane contacts was significantly lower than in old rats. No age-related differences in the viscosity of lipid-lipid membrane contacts and in the polarity of protein-lipid contacts and lipid layers were found. Liver cirrhosis induced by carbon tetrachloride and ethanol administration was associated with increased fluidity of the plasma and mitochondrial membranes of hepatocytes in rats of both age groups. The decrease in membrane viscosity in young rats occurred due to a decrease of the viscosity in the area of protein-lipid and lipid-lipid contacts, while in old rats in the area of protein-lipid contacts. Carbon tetrachloride and ethanol did not affect the polarity of lipid contacts and lipid layers.
Subject(s)
Carbon Tetrachloride/toxicity , Ethanol/toxicity , Hepatocytes/drug effects , Liver Cirrhosis, Experimental/metabolism , Liver/drug effects , Age Factors , Animals , Cell Membrane/chemistry , Cell Membrane/drug effects , Hepatocytes/metabolism , Hepatocytes/pathology , Liver/metabolism , Liver/pathology , Liver Cirrhosis, Experimental/chemically induced , Liver Cirrhosis, Experimental/pathology , Male , Mitochondria/chemistry , Mitochondria/drug effects , Mitochondrial Membranes/chemistry , Mitochondrial Membranes/drug effects , Rats , Rats, Wistar , Viscosity/drug effectsABSTRACT
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.
Subject(s)
Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells , Animals , Carbon Tetrachloride/toxicity , Endothelial Cells , Hepatocytes/pathology , Liver/metabolism , Liver Cirrhosis/metabolism , Male , Rats , Rats, WistarABSTRACT
We studied the formation of injuries in lung endothelium and the response of angiogenesis cells during modeling of pulmonary emphysema in male and female C57BL/6 mice with metabolic disorders. Hemodynamic disturbances and reduction in the area of the microvasculature caused by combined pathology in male mice were more pronounced than in females. Mobilization and migration of angiogenic precursors were impaired in both male and female mice. In males, activity of recruiting endothelial progenitor cells, vascular smooth muscle cells, luminal cells of nascent vessels and pericytes into the lungs was additionally reduced. In females, accumulation of endothelial progenitor cells (CD45-CD31+CD34+), vascular smooth muscle cells, and pericytes in the lungs was observed, which indicated activation of endothelial regeneration. Sex differences in the reaction of the lung endothelium and angiogenesis cells can be explained by genetic factors of lipid and glucose metabolism.
Subject(s)
Endothelium/metabolism , Endothelium/pathology , Lung/metabolism , Lung/pathology , Pulmonary Emphysema/metabolism , Pulmonary Emphysema/pathology , Animals , Antigens, CD34/metabolism , Dyslipidemias/metabolism , Dyslipidemias/pathology , Female , Hyperglycemia/metabolism , Hyperglycemia/pathology , Leukocyte Common Antigens/metabolism , Male , Mice , Mice, Inbred C57BL , Platelet Endothelial Cell Adhesion Molecule-1/metabolism , Sex CharacteristicsABSTRACT
We studied the effects of spiperone, a selective blocker of dopamine D2 receptors, on the model of pulmonary emphysema provoked by administration of elastase and D-galactosamine hydrochloride to female C57BL/6 mice and characterized by activation of proteases in the lungs and systemic deficiency of its inhibitor α1-antitrypsin. In this model, spiperone prevented the development of inflammatory reaction and reduced the area of emphysematous expanded alveolar tissue. The expression of angiogenic marker CD31 in the lungs increased under these conditions. Regeneration of the damaged microvascular bed under the action of spiperone resulted from recruiting of Notch1+ endothelial progenitor cells (CD45-CD31+CD34+) into the lungs and blockade of the inhibitory effect of dopamine on phosphorylation of VEGF-2 receptors in endothelial cells of different maturity. In addition, spiperone produced a protective effect on hepatocytes and restored the production and secretion of α1-antitrypsin by these cells.
Subject(s)
Dopamine Antagonists/pharmacology , Endothelial Progenitor Cells/drug effects , Pulmonary Emphysema/drug therapy , Receptor, Notch1/genetics , Receptors, Dopamine D2/genetics , Spiperone/pharmacology , alpha 1-Antitrypsin Deficiency/drug therapy , Animals , Endothelial Progenitor Cells/metabolism , Endothelial Progenitor Cells/pathology , Female , Galactosamine/administration & dosage , Gene Expression Regulation , Hepatocytes/drug effects , Hepatocytes/metabolism , Hepatocytes/pathology , Liver/drug effects , Liver/metabolism , Liver/pathology , Lung/drug effects , Lung/metabolism , Lung/pathology , Mice , Mice, Inbred C57BL , Neovascularization, Physiologic/drug effects , Pancreatic Elastase/administration & dosage , Phosphorylation/drug effects , Platelet Endothelial Cell Adhesion Molecule-1/genetics , Platelet Endothelial Cell Adhesion Molecule-1/metabolism , Pulmonary Emphysema/chemically induced , Pulmonary Emphysema/genetics , Pulmonary Emphysema/metabolism , Receptor, Notch1/agonists , Receptor, Notch1/metabolism , Receptors, Dopamine D2/metabolism , Regeneration/drug effects , Vascular Endothelial Growth Factor Receptor-2/genetics , Vascular Endothelial Growth Factor Receptor-2/metabolism , alpha 1-Antitrypsin/genetics , alpha 1-Antitrypsin/metabolism , alpha 1-Antitrypsin Deficiency/enzymology , alpha 1-Antitrypsin Deficiency/genetics , alpha 1-Antitrypsin Deficiency/pathologyABSTRACT
The changes in endothelial progenitor cells and progenitor cells of angiogenesis, pericytes and smooth muscle cells, were studied in female C57BL/6 mice with a combination of metabolic impairments induced by injections of sodium glutamate and lung emphysema modeled by the administration of cigarette smoke extract. It was observed that sodium glutamate significantly enhances pathological changes in the lungs (inflammation and lung emphysema) induced by the administration of cigarette smoke extract. Recruiting of endothelial progenitor cells (CD45-CD31+CD34+ and CD31+CD34+CD146-) and progenitor cells of angiogenesis (CD45-CD117+CD309+) was registered in the injured lungs. Angiogenesis impairment induced by combined exposure is related to altered migration of pericytes (CD31-CD34-CD146+) and smooth muscle cells (CD31-CD34+CD146+) in emphysema-like enlarged lung tissue.
Subject(s)
Myocytes, Smooth Muscle/cytology , Myocytes, Smooth Muscle/metabolism , Pericytes/cytology , Pericytes/metabolism , Pulmonary Emphysema/metabolism , Pulmonary Emphysema/pathology , Animals , Antigens, CD34/metabolism , CD146 Antigen/metabolism , Cigarette Smoking/adverse effects , Endothelial Progenitor Cells/cytology , Endothelial Progenitor Cells/drug effects , Endothelial Progenitor Cells/metabolism , Female , Leukocyte Common Antigens/metabolism , Mice , Mice, Inbred C57BL , Myocytes, Smooth Muscle/drug effects , Platelet Endothelial Cell Adhesion Molecule-1/metabolism , Proto-Oncogene Proteins c-kit/metabolismABSTRACT
The title of the article should read:"Role of ß Cell Precursors in the Regeneration of Insulin-Producing Pancreatic ß Cells under the Influence of the Pegylated Form of Glucagon-Like Peptide 1".
ABSTRACT
We studied the effects of elastase, cigarette smoke extract, D-galactosamine hydrochloride, and tyrosine kinase inhibitor SU5416 on endothelial progenitor cells and angiogenesis precursors, as well as on Notch-1 expression by immature endothelial cells. Simultaneously with pulmonary emphysema, different damaging factors with diverse mechanisms of action caused pathological changes in the microvascular network of the lungs and destroyed the alveolar endothelium in female C57Bl/6 mice. D-galactosamine hydrochloride disturbed mobilization of endothelial progenitor cells expressing VEGFR (CD45-CD309+) and angiogenesis progenitors (CD45-CD309+CD117+) and their migration into emphysema expanded lungs. Elastase inhibited VEGFR-expressing endothelial progenitor cells, while cigarette smoke extract inhibited cells with CD45-CD31+CD34+ phenotype. In pulmonary emphysema provoked by elastase or D-galactosamine hydrochloride, angiogenesis was provided by endothelial cells with CD45-CD31+CD34+ phenotype, whereas in emphysema modeled with SU5416 or cigarette smoke extract, it was provided by the endothelial VEGFR-expressing cells and mature CD31+ endothelial cells, respectively. Replenishment of immature endothelial cells damaged by elastase and SU5416 involved Notch-1+ angiogenesis precursors and Notch-1+ endothelial progenitor cells with VEGFR.
Subject(s)
Endothelial Progenitor Cells/cytology , Neovascularization, Physiologic , Pulmonary Emphysema/metabolism , Receptor, Notch1/genetics , Regeneration/physiology , Signal Transduction , Animals , Antigens, CD/genetics , Antigens, CD/metabolism , Complex Mixtures/isolation & purification , Complex Mixtures/toxicity , Endothelial Progenitor Cells/metabolism , Endothelium/cytology , Endothelium/metabolism , Female , Galactosamine/toxicity , Gene Expression Regulation , Indoles/toxicity , Lung/drug effects , Lung/metabolism , Lung/pathology , Mice , Mice, Inbred C57BL , Pancreatic Elastase/toxicity , Pulmonary Emphysema/chemically induced , Pulmonary Emphysema/genetics , Pulmonary Emphysema/pathology , Pyrroles/toxicity , Receptor, Notch1/metabolism , Nicotiana/chemistry , Nicotiana/toxicity , Vascular Endothelial Growth Factor Receptor-1/genetics , Vascular Endothelial Growth Factor Receptor-1/metabolismABSTRACT
The effects of the pegylated form of glucagon-like peptide 1 (pegGLP-1) on oligopotent ß cell precursors (CD45-TER119-CD133+CD49flow) in the pancreas were studied in C57Bl/6 mice. Under conditions of streptozotocin-induced type 1 diabetes mellitus, intraperitoneal injection of pegGLP1 increased the content of ß cell precursors and dithizone-stained cells in the pancreas. ß Cell precursors of mice with diabetes demonstrated high self-maintenance potential. In contrast to pegGLP-1, native GLP-1 did not affect ß cell precursors in diabetic animals. Treatment of a culture of ß cell precursors from mice with diabetes induced the yield of dithizone-stained mononuclears. In conditioned mediums of dithizone-positive cells obtained as a result of differentiation of ß cell precursors from mice with diabetes, insulin was detected after administration of pegGLP-1 (10-7 M) and glucose (3 mmol/liter); the level of insulin increased with increasing glucose concentration (to 20 mmol/liter). The in vitro effect of pegGLP-1 did not differ from the effect of GLP-1 (10-7 M).
Subject(s)
Diabetes Mellitus, Experimental/drug therapy , Glucagon-Like Peptide 1/pharmacology , Incretins/pharmacology , Insulin-Secreting Cells/drug effects , Insulin/agonists , Polyethylene Glycols/chemistry , Animals , Antigens, CD/metabolism , Biomarkers/metabolism , Cell Differentiation/drug effects , Diabetes Mellitus, Experimental/chemically induced , Diabetes Mellitus, Experimental/metabolism , Diabetes Mellitus, Experimental/pathology , Glucagon-Like Peptide 1/analogs & derivatives , Incretins/chemistry , Injections, Intraperitoneal , Insulin/biosynthesis , Insulin-Secreting Cells/metabolism , Insulin-Secreting Cells/pathology , Male , Mice , Mice, Inbred C57BL , Regeneration , StreptozocinABSTRACT
Course administration streptozotocin to male C57Bl/6 mice induces a complex of symptoms typical of type 1 diabetes mellitus: hyperglycemia and insulin deficiency, focal inflammatory infiltration of the pancreas, destructive changes in the Langerhans islets, damage to the insular apparatus (reduced number of PDX1+ cells and insulin expression by the secreting cells). Male reproductive disorder are serious complications of type 1 diabetes mellitus. In "diabetic" mice, interstitial edema with inflammatory infiltration and microvascular disorders in the testicular tissue are observed, the number of endothelial precursors (CD45-/CD31+) and the total number and percentage of motile spermatozoa decreased, immature spermatogenic epithelium cells are desquamated of into the lumen of the tubules. Disturbances in the proliferation and differentiation of various spermatogonial stem cell populations (c-kit-/CD90+, c-kit+/CD90+, and CD51-/CD24+/CD52+) in diabetes can be explained by the inhibitory influence of inflammatory factors on testosterone-producing Leydig cells.
Subject(s)
Diabetes Mellitus, Experimental/pathology , Erectile Dysfunction/pathology , Leydig Cells/drug effects , Oligospermia/pathology , Sertoli Cells/drug effects , Streptozocin/toxicity , Animals , Antigens, CD/genetics , Antigens, CD/metabolism , Cell Count , Cell Differentiation/drug effects , Cell Movement , Cell Proliferation/drug effects , Diabetes Mellitus, Experimental/chemically induced , Diabetes Mellitus, Experimental/genetics , Diabetes Mellitus, Experimental/metabolism , Diabetes Mellitus, Type 1/genetics , Diabetes Mellitus, Type 1/metabolism , Diabetes Mellitus, Type 1/pathology , Endothelial Progenitor Cells/drug effects , Endothelial Progenitor Cells/metabolism , Endothelial Progenitor Cells/pathology , Erectile Dysfunction/chemically induced , Erectile Dysfunction/genetics , Erectile Dysfunction/metabolism , Gene Expression , Homeodomain Proteins/genetics , Homeodomain Proteins/metabolism , Insulin/genetics , Insulin/metabolism , Islets of Langerhans/drug effects , Islets of Langerhans/metabolism , Islets of Langerhans/pathology , Leydig Cells/metabolism , Leydig Cells/pathology , Male , Mice , Mice, Inbred C57BL , Oligospermia/chemically induced , Oligospermia/genetics , Oligospermia/metabolism , Sertoli Cells/metabolism , Sertoli Cells/pathology , Spermatogenesis/drug effects , Spermatogenesis/genetics , Spermatogonia/drug effects , Spermatogonia/metabolism , Spermatogonia/pathology , Spermatozoa/drug effects , Spermatozoa/metabolism , Spermatozoa/pathology , Trans-Activators/genetics , Trans-Activators/metabolismABSTRACT
Biological activity of a new pegylated form of an of glucagon-like peptide-1 (GLP-1) analogue pegGLP-1 was studied in C57Bl/6 mice under normal conditions and during modeling of streptozotocin-induced type I diabetes mellitus. pegGLP-1 differs from GLP-1 (7-37) by polyethylene glycol residue covalently bound to His7, Lys26, and Lys34 of the GLP-1 molecule. It was shown that single intragastrical administration of pegGLP-1 induced an increase in GLP-1 level in blood serum of healthy mice. The maximum level of this parameter was observed in 4-8 h. pegGLP-1 elimination half-time was 8.5 h and mean retention time was 15 h. Administration of pegGLP-1 to animals with modeled type I diabetes mellitus was followed by an increase in the levels of GLP-1 and insulin in blood serum, produced a hypoglycemic effect, and improved the parameters of glucose-tolerance test. Biological activity of pegGLP-1 was higher than activity of GLP-1.
Subject(s)
Diabetes Mellitus, Experimental/drug therapy , Glucagon-Like Peptide 1/analogs & derivatives , Glucagon-Like Peptide 1/therapeutic use , Animals , Blood Glucose/drug effects , Diabetes Mellitus, Experimental/blood , Glucagon-Like Peptide 1/blood , Hypoglycemic Agents/therapeutic use , Insulin/blood , Insulin/therapeutic use , Male , Mice , Mice, Inbred C57BLABSTRACT
The properties of spermatogonial stem cells, endothelial progenitor cells, and the epithelial progenitors of C57Bl/6 mice under conditions of metabolic disorders were studied using the model of busulfan-induced suppression of spermatogenesis and in vitro culture technique. Spermatogonial stem cells CD117-CD90+ and epithelial progenitors CD45-CD31-Sca-1+CD49f+ derived from the testes of mice with metabolic disturbances demonstrated 17- and 28-fold increase in the respective cell mass and generated cell colonies in vitro. In contrast, spermatogonial stem cells with immune phenotype CD51-CD24+CD52+ had reduced selfrenewal capacity. Spermatogonial stem cells CD117-CD90+ and CD117+CD90+ as well as endothelial progenitors CD45-CD31+ derived from the testes of donor mice with metabolic disorders demonstrated high transplantation capacity in C57Bl/6 mouse testes damaged by cytostatic busulfan.
Subject(s)
Endothelial Progenitor Cells/cytology , Stem Cells/cytology , Animals , Busulfan/pharmacology , CD24 Antigen/metabolism , CD52 Antigen/metabolism , Endothelial Progenitor Cells/drug effects , Inflammation/metabolism , Integrin alphaV/metabolism , Male , Metabolic Diseases/metabolism , Mice , Mice, Inbred C57BL , Proto-Oncogene Proteins c-kit/metabolism , Spermatogenesis/drug effects , Spermatogonia/cytology , Spermatogonia/drug effects , Stem Cells/drug effects , Testis/drug effects , Testis/metabolism , Thy-1 Antigens/metabolismABSTRACT
The regenerative potential of stem and progenitor cells from ischemic testes of C57Bl/6 mice was studied in vitro (cell culture) and in vivo (mouse model of busulfan-induced suppression of spermatogenesis). Spermatogonial stem cells with phenotypes CD117-CD90+ and CD51-CD24+CD52+ from ischemic testes demonstrated 33-fold and 7-fold increments of cell mass and generated colonies in vitro. Epithelial (CD45-CD31-Sca-1+CD49f+) and endothelial (CD45-CD31+) precursors exhibited lower self-renewal capacity. On day 30 after injection of stem and progenitor cells from ischemic testes to the rete testis zone of the testes of busulfantreated animals, an increase in the count of CD117-CD90+ spermatogonial stem cells, total count, and mobile sperm count in the testes of recipient mice was observed. In addition, we observed an increase in Sca-1+ cell count, recovery of the spermatogenic epithelium in the seminiferous tubules, and appearance of immature Leydig cells in "busulfan" testes; the level of tissue testosterone and fertility index also increased.
Subject(s)
Busulfan/toxicity , Ischemia/metabolism , Mesenchymal Stem Cells/drug effects , Regeneration/drug effects , Spermatogenesis/drug effects , Spermatogonia/metabolism , Animals , Antigens, CD/genetics , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Endothelial Cells/drug effects , Endothelial Cells/metabolism , Endothelial Cells/pathology , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Epithelial Cells/pathology , Gene Expression , Ischemia/pathology , Leydig Cells/drug effects , Leydig Cells/metabolism , Leydig Cells/pathology , Ligation , Male , Mesenchymal Stem Cells/metabolism , Mesenchymal Stem Cells/pathology , Mice , Mice, Inbred C57BL , Seminiferous Tubules/drug effects , Seminiferous Tubules/metabolism , Seminiferous Tubules/pathology , Spermatic Cord/blood supply , Spermatic Cord/surgery , Spermatogonia/drug effects , Spermatogonia/pathology , Stem Cell Transplantation , Testosterone/biosynthesisABSTRACT
Using the model of hypogonadism in C57Bl/6 male mice, we showed that injection of streptozotocin to newborn animals and high-fat diet induced serum IFN-γ and IL-17 elevation, glucose metabolism disturbances, insulin resistance, destructive changes of the Langerhans islets (deficit of PDX1+ß cells), while the number of oligopotent ß cell precursors (CD45-TER119-CD133+CD49flow) increased. Diabetes played the role of an inducer of testicular tissue inflammation (pan-hemopoietic cell infiltration, increase of IL-2, IL-17, and IL-23 content) and reproductive system disturbances in mice (decrease in free testosterone concentration, suppression of spermatogenesis, and infertility). The development of hypogonadism was paralleled by an increase in the count of spermatogonial stem cells (CD117+CD29+CD90+), multipotent mesenchymal stromal cells (CD45-CD31-CD90+CD106+), hemangiogenesis precursors (CD45-CD117+Flk1+), and epithelial cells (CD45-CD31-CD49f+CD326+).
Subject(s)
Diabetes Mellitus, Experimental/pathology , Hypogonadism/pathology , Pancreas/pathology , Regeneration/immunology , Testis/pathology , Animals , Antigens, CD/genetics , Antigens, CD/immunology , Blood Glucose/immunology , Blood Glucose/metabolism , Diabetes Mellitus, Experimental/chemically induced , Diabetes Mellitus, Experimental/genetics , Diabetes Mellitus, Experimental/immunology , Diet, High-Fat , Female , Gene Expression , Hypogonadism/chemically induced , Hypogonadism/genetics , Hypogonadism/immunology , Immunophenotyping , Insulin-Secreting Cells/immunology , Insulin-Secreting Cells/pathology , Interferon-gamma/genetics , Interferon-gamma/immunology , Interleukin-17/genetics , Interleukin-17/immunology , Interleukin-2/genetics , Interleukin-2/immunology , Interleukin-23/genetics , Interleukin-23/immunology , Male , Mice , Mice, Inbred C57BL , Organ Specificity , Pancreas/immunology , Regeneration/genetics , Spermatogenesis/genetics , Spermatogonia/immunology , Spermatogonia/pathology , Stem Cells/immunology , Stem Cells/pathology , Streptozocin , Testis/immunologyABSTRACT
Stem and progenitor cells were studied on mouse model of testicular ischemia. Testicular ischemia led to a decrease in free testosterone concentration. Hemodynamic changes, interstitial edema, and destruction of spermatogenic epithelium, Leydig, and Sertoli cells were observed in the testicular tissue. Accumulation of degenerative germ cells was accompanied by reduction in the count of spermatogonial stem cells with immunophenotype CD117(-)CD29(+)CD90(+) and CD117(+)CD29(+)CD90(+). Simultaneously with pathomorphological changes in the testes and suppression of spermatogenesis, ischemia reduced the count of hematopoietic progenitor cells, hematopoietic stem cells with immunophenotype Lin(-)CD117(+)Sca-1(+)c-kit(+)CD34(+) and Lin(-)CD117(+)Sca-1(+)c-kit(+)CD34(-), and multipotent mesenchymal stromal cells (CD45(-)CD31(-) CD90(+)CD106(+)) in the testicular tissue. The population of CD45(-)CD31(+)-endothelial cells in ischemic testicular tissue increased.
Subject(s)
Endothelial Cells/pathology , Hematopoietic Stem Cells/pathology , Ischemia/pathology , Mesenchymal Stem Cells/pathology , Testis/cytology , Testis/pathology , Animals , Antigens, CD34/metabolism , Antigens, Ly/metabolism , Cell Differentiation/physiology , Disease Models, Animal , Endothelial Cells/metabolism , Hematopoietic Stem Cells/metabolism , Ischemia/metabolism , Male , Membrane Proteins/metabolism , Mesenchymal Stem Cells/metabolism , Mice , Proto-Oncogene Proteins c-kit/metabolism , Testis/metabolismABSTRACT
Inflammation, extracellular matrix proteins (hydroxyproline, connective tissue growth factor, collagen, and fibronectin), stem and progenitor cells (multipotent mesenchymal stromal cells, Clara cells, angiogenesis, precursors, endothelial and epithelial cells) were studied in female C57Bl/6 mice with experimental elastase-induced emphysema. Diffuse emphysema reduced the number of endothelial (CD45(-)CD31(+)CD34(+)) and epithelial (CD45(-)CD117(+)CD49f(+)) cells, induced microcirculation disturbances, and decreased the area occupied by the connective tissue. Emphysematous changes in the lungs were accompanied by infiltration of the alveolar septa with macrophages and lymphocytes, increase in the serum and lung concentrations of transforming growth factor-ß, IL-1ß, IL-2, IL-5, IL-10, and IL-13, and lung concentration of IL-17. In the lungs, inflammation was associated with marked increase in the number of multipotent mesenchymal stromal cells CD90(+)CD73(+)CD106(+)CD44(+)) and Clara cells (CD45(-)CD34(-)CD31(-)Sca1(+)) and overexpression of extracellular matrix proteins (hydroxyproline, connective tissue growth factor, collagen, fibronectin) and Clara cells protein. On the other hand, elastase reduced the number of angiogenic precursor cells (CD45(-)CD117(+)Flk1(+)).
