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비성빈혈에 있어서의 Erythropoietin 활성도에 관한 연구

Other Titles
 Study for erythropoietin activity in splenic anemia 
Issue Date
1975
Description
의학과/박사
Abstract
[한글] 비장기능항진시에 유발되는 빈혈의 원인을 추구하고자, methylcellulose를 백서에 반복투여함으로서 실험적 비기능항진증을 유발시킨 다음, 출혈성빈혈군과 혈액학적으로 비교 관찰함과 아울러, 이들의 적혈구, 골수 및 비장에서의 erythrepoietin 활성도를 측정하고, 동시에 사람에서도 정상인 및 비기능항진증 환자에서 erythropoietin활성도를 측정하여 다음과 같은 결과를 얻었다. Methylcellulose 투여로 현저한 비종대와 함께 빈혈 및 혈소판 감소가 초래되었으나 백혈구의 변화는 별로 없었다. 망상구는 출혈성빈혈군에서 대조군에 비해 현저히 증가되었으며 비성빈혈군에서는 대조군에 비해 증가는 되어 있었으나 출혈성 빈혈군에 비하여 낮은치를 보였다. 실험동물군에서 적혈구의 erythropoietin의 활성도는 출혈성빈혈군에서 대조군에 비하여 통계학적으로 의의있는 증가를 보였으나 비성빈혈군에서는 대조군에 비하여 증가는 하였으나 통계학적 의의는 없었다. 사람에서는 비성빈혈군의 erythropoietin 활성도가 대조군에 비하여 현저히 낮았다. 골수 및 비장에서의 erythropoietin 활성도는 실험동물군에서는 비성빈혈군이 대조군보다 낮은 치를 나타내고 있으며 사람에서는 의의있게 감소하였다. 이상과 같은 결과를 종합검토하면 비기능항진증에 있어서 빈혈의 성인의 일부는 골수에서의 조혈기능이 억제되는데 기인된다고 추리되며, 이러한 억제작용이 erythropeietin의 분비가 감소됨으로서 기인된 것인지, erythropoietin의 분비는 정상이나 종대된 비장에서 유리되는 소위 humoral factor에 의한 것인지는 앞으로 더욱 추구하여야 할 문제라 사료된다. Study for Erythropoietin Activity in Splenic Anemia Jee Sook Hahn, M.D. Department of Medical Science, The Graduate School, Yonsei University (Directed by Prof. Eung Suk Chai, M.D., D.M. Sc.) The syndrome of hypersplenism is characterized by splenomegaly, selective or total blood cytopenias, normal or hyperplastic bone marrow and disappearence of the cytopenias after splenectomy. The mechanism of the various forms of hypersplenism is still under discussion. Clinical or experimental evidence has been presented in favor of two different hypothesis, the destruction of blood elements by sequestration in the spleen, and the inhibition of maturation and/or liberation of bone marrow cells by humoral mechanism. Therefore, the present investigation was undertaken to study the effects of erythropoietin on Fe**59 uptake by red cell, bone marrow, and spleen-of control, normal bled anemia, and methylcellulose induced splenic anemia group in albino rats: control and splenic anemia group in human beings-to provide evidences which may be helpful in defining the mechanism of the splenic anemia Materials and Methods A. Experimental groups: Albino rats, 180-200gm in weight, regardless of sex, were divided into three groups: control, bled anemic group and methylcellulose-induced hypersplenic group, and each group was consisted of 5 albino rats. To prepare methylcellulose anemic group by Palmar's method (1953), methylcellulose of a viscosity grade of 400 centipoises was obtained as a dry powder, and dissolved by slowly adding 2.5gm to 100m1 of distilled water at a temperature of approximately 80℃ with constant stirring. The mixture was then allowed to cool and a clear viscous solution formed which was refrigerated until used. Two ml of the methylcellulose solution were injected intraperitoneally twice weekly through an 18 gauge needle into the albino rats. After 10-12 weeks, hemoglobin, leukocyte, platelet determination and reticulocyte counts with the blood obtaining from the tail vein were made. Bled anemic groups were made by continuous bleeding 1-1.5ml of blood every day through tail vein of albino rats for 2-3 weeks. Hemoglobin, leukocyte, platelet and reticulocyte were also measured after that period. B. Human groups: Five cases were respectively selected as a normal adult control and human splenic groups. Five cartes of splenic group were clinically confirmed for the hypersplenism. C. Measurement of erythropoietin activity : The actual assay for erythropoietin is interpreted to calculate the percent of the tracer dose of Fe**59 into the circulating erythrocyte mass, using Fried's method (1957). During five days of experimental period, the albino rats were in starvation state permitting only water, and 1ml of teat serum whs administered intraperitoneally on the 2nd and 3rd day of the experiment respectively. For the determination of radioiron incorporation, 1μCi of Fe**59 as ferrous citrate was injected intraperitoneally on the 4th day. and 1ml. of blood sample was obtained by cardiac punture on the fifth day-18 hours after Fe**59 administration. Its radioactivity was assayed in a well-type gamma detector and the per cent of the dose of Fe**59 present in circulating erythrocytes was calculated as fellows: Fe**59 uptake in circulating erythrocytes (%/ml.) = (net couts/Jml. bloods X 0.05 X body weight) / (net counts injected) X 100 And after femoral vein was exposed under ether anesthesia, it was irrigated with physiologic saline via inferior vena cava. Extirpation of spleen and right femur was dane. Their radioactivity was assayed and calculated by the following formula: Bone marrow or spleen Fe**59 uptake(%/gm.) = (net counts/gm of bone marrow or spleen) / (net counts injected) X 100 Results and Conclusions The results are summarized as follows: 1. Repeated administration of methylcellulose into albino rata induced splenomegaly. anemia and thrombocytopenia, whereas leukocyte was slightly depressed with mild reticulocytosis. 2. Slightly elevated value of reticulocyte was revealed in methylcellulose induced splenic group but its level was shown to be markedly lower than that of bled-anemic group. 3. In group which injected serum of albino rats and human beings, the red cell Fe**59 uptake was decreased in splenic anemia comparing with normal bled anemia and normal human control group respectively. 4. The bone marrow and spleen Fe**59 uptake of splenic anemia was slightly lower than that of control in rats, but it was significantly decreased in splenic anemia group of human beings. The results obtained from present studs indicate that erythropoietin activity is depressed in splenic anemia and support an inhibitory action on erythropoiesis in the bone marrow. However, it should be searched for further investigation whether thin inhibitory effect may be attributed to decreased release of erythropoietin in splenic anemia, or the erythropoietin activity is suppressed by a certain substance (possible humboral factor) released from the enlarged spleen and present in the serum even though the erythropoietin is normally released.
[영문] The syndrome of hypersplenism is characterized by splenomegaly, selective or total blood cytopenias, normal or hyperplastic bone marrow and disappearence of the cytopenias after splenectomy. The mechanism of the various forms of hypersplenism is still under discussion. Clinical or experimental evidence has been presented in favor of two different hypothesis, the destruction of blood elements by sequestration in the spleen, and the inhibition of maturation and/or liberation of bone marrow cells by humoral mechanism. Therefore, the present investigation was undertaken to study the effects of erythropoietin on Fe**59 uptake by red cell, bone marrow, and spleen-of control, normal bled anemia, and methylcellulose induced splenic anemia group in albino rats: control and splenic anemia group in human beings-to provide evidences which may be helpful in defining the mechanism of the splenic anemia Materials and Methods A. Experimental groups: Albino rats, 180-200gm in weight, regardless of sex, were divided into three groups: control, bled anemic group and methylcellulose-induced hypersplenic group, and each group was consisted of 5 albino rats. To prepare methylcellulose anemic group by Palmar's method (1953), methylcellulose of a viscosity grade of 400 centipoises was obtained as a dry powder, and dissolved by slowly adding 2.5gm to 100m1 of distilled water at a temperature of approximately 80℃ with constant stirring. The mixture was then allowed to cool and a clear viscous solution formed which was refrigerated until used. Two ml of the methylcellulose solution were injected intraperitoneally twice weekly through an 18 gauge needle into the albino rats. After 10-12 weeks, hemoglobin, leukocyte, platelet determination and reticulocyte counts with the blood obtaining from the tail vein were made. Bled anemic groups were made by continuous bleeding 1-1.5ml of blood every day through tail vein of albino rats for 2-3 weeks. Hemoglobin, leukocyte, platelet and reticulocyte were also measured after that period. B. Human groups: Five cases were respectively selected as a normal adult control and human splenic groups. Five cartes of splenic group were clinically confirmed for the hypersplenism. C. Measurement of erythropoietin activity : The actual assay for erythropoietin is interpreted to calculate the percent of the tracer dose of Fe**59 into the circulating erythrocyte mass, using Fried's method (1957). During five days of experimental period, the albino rats were in starvation state permitting only water, and 1ml of teat serum whs administered intraperitoneally on the 2nd and 3rd day of the experiment respectively. For the determination of radioiron incorporation, 1μCi of Fe**59 as ferrous citrate was injected intraperitoneally on the 4th day. and 1ml. of blood sample was obtained by cardiac punture on the fifth day-18 hours after Fe**59 administration. Its radioactivity was assayed in a well-type gamma detector and the per cent of the dose of Fe**59 present in circulating erythrocytes was calculated as fellows: Fe**59 uptake in circulating erythrocytes (%/ml.) = (net couts/Jml. bloods X 0.05 X body weight) / (net counts injected) X 100 And after femoral vein was exposed under ether anesthesia, it was irrigated with physiologic saline via inferior vena cava. Extirpation of spleen and right femur was dane. Their radioactivity was assayed and calculated by the following formula: Bone marrow or spleen Fe**59 uptake(%/gm.) = (net counts/gm of bone marrow or spleen) / (net counts injected) X 100 Results and Conclusions The results are summarized as follows: 1. Repeated administration of methylcellulose into albino rata induced splenomegaly. anemia and thrombocytopenia, whereas leukocyte was slightly depressed with mild reticulocytosis. 2. Slightly elevated value of reticulocyte was revealed in methylcellulose induced splenic group but its level was shown to be markedly lower than that of bled-anemic group. 3. In group which injected serum of albino rats and human beings, the red cell Fe**59 uptake was decreased in splenic anemia comparing with normal bled anemia and normal human control group respectively. 4. The bone marrow and spleen Fe**59 uptake of splenic anemia was slightly lower than that of control in rats, but it was significantly decreased in splenic anemia group of human beings. The results obtained from present studs indicate that erythropoietin activity is depressed in splenic anemia and support an inhibitory action on erythropoiesis in the bone marrow. However, it should be searched for further investigation whether thin inhibitory effect may be attributed to decreased release of erythropoietin in splenic anemia, or the erythropoietin activity is suppressed by a certain substance (possible humboral factor) released from the enlarged spleen and present in the serum even though the erythropoietin is normally released.
URI
http://ir.ymlib.yonsei.ac.kr/handle/22282913/117334
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2. 학위논문 > 1. College of Medicine (의과대학) > 박사
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