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  • Proliferacijska sposobnost hematopoetskih matičnih stanica kod kroničnog bubrežnog zatajenja : doktorska disertacija
    Glaser, Marjana
    Chronic renal failure (CRF) is accompanied by numerous complications including hematologic, and among these anemia is the most obvious. It is assumed that apart from the known causes such as the lack ... of erythropoietina (EPO), the entire hematopoesis is also disturbed. Numerous substances are involded in the inhibition of erythropoiesis. However, it remains unelucidated at which level and in which part of hematopoietic stem cells and by which mechanism inhibition occurs and whether this phenomenon is the exclusive result of the lack of EPO. Therefore the aim of this study was to verify the hypothesis that, apart from the lack of EPO, other agents also play a part in the pathogenesis of anemia in CRF, and whether this disturbance affects the entire hematopoiesis and at a higher hierarchic level of stem cells than that of erytrocytopoiesis. So the basic aim of the study was to investigate the clonogenic and proliferative capacity not merely of stem cells for erythrocytopoiesis (BFU E), but the clonogenic and proliferative capacity of cells whose development is not affected by EPO (granulocyte-monocyte colony forming unit - CFU GM), or of several types of cells (colony forming unit granulocyte-erithrocyte-megakaryocyte CFU GEMM) whose development is also not affected by EPO. The aim was to test their functional capacity to respond to the regulatory stimulus in the culture. In this way we wanted to establish to what extent and which part of hematopoiesis is inhibited in CRF patients as a possible cause of anemia. The specific aims of this study were: to determine the cologenic and proliferative capacity of the multipotent cell for myelopoiesis (CFU GEMM), the target stem cell for granulocytopoiesis and monocytopoiesis (CFU GM) and the stem cell for erythrocytopoiesis (BFU E), as well as to determine EPO concentration and evaluate its relation to the proliferative capacity of stem cells. In our study, statistical analysis of 21 parameters was done in a population of 65 CRF patients and a Control group of 30 healthy subjects. With respect to serum creatinine concentrations, the patients were divided in two groups: Group 1 with creatinine < 900 micromol/L included 24 patients, Group 2 with creatinine > 900 micromol/L included 41 patients. The patients were treated with maintenance hemodialysis. During the last 21 days they received no blood transfusion and so far they had received no human erythropoietin, but the did show signs of normocytic normochromic anemia. The average age of the patients was 55.33 +/- 13.15 years (from 20 to 79), in the Control group consisting of 15 women and 15 men it was 50.03 +/- 13.71 years (from 20 to 76). Standard methods were used to analyse: creatinine, urea, uric acid, sodium, potassium and calcium for the evaluation of renal function, the erithrocyte, leukocyte and thrombocyte counts, myelogram, hemoglobin, mean cell volumne, iron, TIBC, ferritin, erythrocyte sedimentation for the evaluation of hematopoiesis. The clonogenic and proliferative capacity of stem cells was deteremined using the in vitro method of short-term cultivation of cells from peripheral blood. EPO concentration was assessed with the ELISA method. Parametric statistical methods and a comparison of the growth of stem cell colonies in Group 1 and 2 were used to establish a statistically significant decrease in the growth of colonies CFU GEMM in Group 2 (p<0.002). Comparing group 1 with the Control group showed no statistically significant difference in the growth of colonies BFU E, CFU GM and CFU GEMM and in EPO concentration. Comparing Group 2 and the Control group, a statistically significant decrease in the growth of colonies BFU E (p<0.002) and CFU GM (p<0.05) was established. Comparing the entire group of patients and the Control group, a statistically significant decrease in the growth of colonies BFU E (p<0,001) and CFU GM (p<0.04) was established, with a smaller number of colonies in the population of patients. The method of correlation was used to investigate the interaction of individual laboratory parameters with the number of stem cell colonies. In Group 1, the number of grown colonies CFU GEMM was in positive correlation with the number of grown colonies BFU E (p<0.001) and CFU GM (p<0.001) and the number of grown colonies CFU GM in positive correlation with number of colonies BFU E (p<0,001). The growth of colonies CFU GM (p<0.02) and BFU E (p<0.03) was in negative correlation with creatinine concentration. The EPO concentration was in negative correlation with creatinine concentration (p<0.02). In Group 2, a positive correlation was found between the number of colonies CFU GM and BFU E (p<0.01), EPO concentration was in negative correlation with erythrocyte sedimentation (p<0.02) and iron concentration (p<0.05). In the entire population of patients, a positive correlation of the growth of colonies CFU GEMM with BFU E (p<0.01) and CFU GM (p<0.01) was established. A negative correlation of the growth of colonies CFU GEMM (p<0.01), CFU GM (p<0.05) and BFU E (p<0.05) with urea concentration was also established. In contrast to previous studies dealing with the growth of colonies CFU E and BFU E from bone marrow or blood with or without the addition of uremic serum, our present study investigated three different groups of stem cells: BFU E, CFU GM and CFU GEMM in blood and EPO concentration in CRF patients, correlating them with the severity of uremia. We confirmed the previous statement that in most CRF patients EPO concentration was normal, however, means a decrease with respect to the grade of uremia. At the same time we established a significant decrease in the growth of colonies BFU E. This lead to the conclusion that the relative decrease in EPO concentration is one of the major causes of anemia in CRF patients, in consideration of the factt that EPO affects the erythropoiesis target cells. We also established a decrease in the growth of stem cell colonies for the growth of other cells such as CFU GM and also immature stem cells for several types of cells CFU GEMM whose development is, as a rule, not affected by EPO. We also found a correlation of the growth of colonies CFU GEMM and CFU GM with the growth of colonies BFU E and a correlation of the growth of CFU GEMM with the degree of advanced uremia, or the concentration of urea and creatinine. Follows the conclusion and confirmation of the hypothesis that the disturbed hematopiesis and the development of anemia in CRF are also affected by other agents not connected with EPO production. They have a negative effect on hematopoiesis as a whole and not merely on erythrocytopoiesis, and they are connected with the degree of anemia. These agents act at a higher level of hematopoiesis development and they have a lasting effect on the stem cells, considering the finding that the growth of colonies was inhibited in vitro despite the addition of growth factors in the culture. Based on these data and those from other studies on the changed immune system, particularly the action of T lymphocytes, the activation of the complement and the function of monocytes in CRF patients, it was hypothesised that the responsibility for the changed hematopoiesis in CRF established in this study rests on the decreased production of stimulatory cytokines (BPA), or the increased production of proinflammatory monokines with inhibitory action. The possible effect of the dialysis procedure itself on these changes is also assumed.
    Type of material - dissertation ; adult, serious
    Publication and manufacture - Zagreb : [M. Glaser], 2002
    Language - croatian
    COBISS.SI-ID - 1079871

Library Call number – location, accession no. ... Copy status
MF, Central Medical Library, Lj. doktorske disertacije
GLASER Marjana Proliferacijska
IN: 020040302 		available - outside loan, loan period: 1 months
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