| Project/Area Number |
16590254
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pathological medical chemistry
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| Research Institution | TOKYO WOMEN'S MEDICAL UNIVERSITY |
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Principal Investigator |
KANNO HITOSHI TOKYO WOMEN'S MEDICAL UNIVERSITY, FACULTY OF MEDICINE, ASSOCIATE PROFESSOR, 医学部, 助教授 (70221207)
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| Co-Investigator(Kenkyū-buntansha) |
AISAKI KEN-ICHI NATIONAL INSTITUTE OF HEALTH AND SCIENCES, DIVISION OF TOXICOLOGY, 毒性部, 研究員 (40322086)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2004: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | hemolytic anemia / inborn error of metabolism / red cell life span / anti-oxidant / 赤血球 / 無効造血 / 変異マウス / 造血細胞コロニー解析 / ピルビン酸キナーゼ / 赤血球分化 / 酸化ストレス |
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| Research Abstract |
We previously established a Friend erythroleukemic cell, SLC3, from the mice model of red blood cell pyruvate kinase (R-PK) deficiency. SLC3 showed spontaneous apoptosis during routine passage. When cultured in a condition of glucose deprivation or supplementation with 2-deoxyglucose, a control Friend cell, CBA2, also showed apoptosis. Preincubation with N-acetyl cysteine, glutathione precursor, reduces apoptosis of CBA2 induced by 2-deoxyglucose, suggesting that glycolytic inhibition increases oxidative stress in erythroid cells and that erythroid apoptosis is likely to be induced by reactive oxygen species (ROS).
When cultured in 5mM phosphoenolpyruvate (PEP) for 48 hours, intracellular concentration of PEP and pyruvate increased up to 7- and 2-times, respectively. ROS in the PEP-treated erythroid cells significantly decreased, and apoptotic cell number decreased about 50% of non-treated cells. This observation suggests that accumulation of PEP improves glycolysis since the mutant R-PK has lower substrate specificity due to the active site mutation. Taken together, we conclude that glycolytic inhibition increases oxidative stress in erythroid cells and activate proapoptotic gene expressions, leading to apoptosis.
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