Ineffective erythropoiesis in mutant mice with deficient pyruvate kinase activity
Ineffective erythropoiesis in mutant mice with deficient pyruvate kinase activity
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DOI：
10.1016/j.exphem.2005.07.008
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发表时间：
2005-11-01
影响因子：
2.6
通讯作者：
Fujii, H
中科院分区：
文献类型：
--
作者：
Aizawa, S;Harada, T;Fujii, H
目标。丙酮酸激酶（PK）缺乏是由糖酵解酶缺陷引起的遗传性非球胞性贫血的最常见原因。在PK缺乏的个体中，由于三磷酸腺苷耗损，红细胞不易变形，并且在网状内皮系统的微循环中被破坏，导致血管外溶血。钾素缺乏症小鼠（PK- i - sic）钾素缺乏症的病理生理机制已被广泛研究。我们用这些小鼠检测了PK缺乏对红细胞祖细胞成熟的影响。材料与方法。采用末端脱氧核苷酸转移酶介导的dUTP镍端标记（TUNEL）染色法观察PK-1（slc）小鼠脾脏凋亡细胞的形态。我们还在体外检测了PK-1（slc）小鼠脾脏中造血干细胞集落的形成，以研究红细胞生成和膜联蛋白V结合，作为构成红细胞集落中凋亡细胞的测量，以评估红细胞祖细胞的成熟。PK-1小鼠脾脏中包括集落形成单位红细胞、爆发形成单位红细胞（BFU-E）、集落形成单位粒细胞-巨噬细胞和多系集落形成单位在内的造血祖细胞数量显著增加，提示造血，尤其是红细胞生成能力增强。TUNEL检测发现PK-1（slc）小鼠脾红髓中存在凋亡细胞。双色流式细胞术检测到抗ter119阳性细胞中的凋亡细胞，提示凋亡细胞属红系。在PK-1（slc）小鼠骨髓细胞生成的BFU-E培养物中检测到凋亡细胞。本研究结果表明，PK缺乏引起的代谢紊乱不仅会改变红细胞的存活，还会影响红细胞祖细胞的成熟，导致红细胞生成无效。(c) 2005年国际实验血液学学会。Elsevier Inc.出版。
Objective. A deficiency of pyruvate kinase (PK) is the most common cause of hereditary nonspherocytic anemia due to glycolytic enzyme defects. Red cells are poorly deformable due to adenosine triphosphate depletion in individuals with a PK deficiency and are destroyed in the microcirculation of the reticuloendothelial system, leading to extravascular hemolysis. The pathophysiology of PK deficiency has been widely studied in PK-deficient mice (PK-I-sIc). We examined the effects of a PK deficiency on erythroid progenitor maturation using these mice.Materials and Methods. The appearance of apoptotic cells in spleen of PK-1(slc) mice was examined by terminal deoxynucleotidyl-transferase-mediated dUTP nick-end labeling (TUNEL) staining. We also assayed hematopoietic stem cell colony formation in vitro in the spleen of PK-1(slc) mice, to investigate erythropoiesis, and annexin V binding, as a measure of apoptotic cells in constitutive erythroid colonies, to evaluate the maturation of erythroid progenitors.Results. The number of hematopoietic progenitors including colony-forming unit erythroids, burst-forming unit erythroids (BFU-E), colony-forming unit granulocyte-macrophages, and multilineage colony-forming units in the spleens of PK-1(slc) was remarkably increased indicating hematopoiesis, and enhanced erythropoiesis in particular. TUNEL assays identified apoptotic cells in the splenic red pulp of the PK-1(slc) mice. Two-color flow cytometry detected apoptotic cells among anti-TER119-positive cells, suggesting that apoptotic cells were of erythroid lineage. Cells undergoing apoptosis were detected in cultures of BFU-E generated from bone marrow cells of PK-1(slc) mice.Conclusions. The results in this study indicate that the metabolic disturbance in PK deficiency alters not only the survival of red cells but also the maturation of erythroid progenitors, resulting in ineffective erythropoiesis. (c) 2005 International Society for Experimental Hematology. Published by Elsevier Inc.