IDENTIFICATION OF THE CAUSATIVE GENES RESPONSIBLE FOR ERYTHROID APOPTOSIS INDUCED BY GLYCOLYTIC INHIBITION

糖酵解抑制诱导的红细胞凋亡的致病基因的鉴定

• 批准号: 16590254
• 负责人: KANNO HITOSHI
• 金额: $ 2.24万
• 依托单位: TOKYO WOMEN'S MEDICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590254/
• 关键词: 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

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.

我们以前从红细胞丙酮酸激酶（R-PK）缺乏的小鼠模型中建立了一个朋友红血病细胞SLC3。 SLC3在常规通过期间显示自发凋亡。当在葡萄糖剥夺或用2-脱氧葡萄糖补充的条件下培养时，对照的朋友细胞CBA2培养也会显示出凋亡。与2-脱氧葡萄糖诱导的CBA2凋亡降低了与N-乙酰半半胱氨酸的预孵育，这表明糖酵解抑制会增加红斑细胞中的氧化应激，从而诱导了脑膜质的氧化应激，并可能通过反应性氧化物（ROSERAI）（ROSERECY cORTINAIR）（ROS）。 （PEP）持续48小时，pep和丙酮酸的细胞内浓度分别增加了7倍和2倍。 PEP处理的红细胞细胞中的ROS显着降低，凋亡细胞数减少了约50％的未处理细胞。该观察结果表明，由于活性位点突变，突变的R-PK的底物特异性较低，因此PEP的积累改善了糖酵解。综上所述，我们得出的结论是，糖酵解抑制会增加红细胞细胞中的氧化应激并激活促凋亡基因表达，从而导致凋亡。

项目成果

Histopathological study of lattice corneal dystrophy with L527R mutation of transforming growth factor-beta induced gene.

转化生长因子-β诱导基因L527R突变引起的格子状角膜营养不良的组织病理学研究。

• 发表时间: 2004
• 期刊: Nippon Ganka Gakkai Zasshi 108
• 作者: Nakagawa E;et al.
• 通讯作者: et al.

Cyclic polylactate inhibited growth of cloned leukemic cells through reducing glycolytic enzyme activities.

环状聚乳酸通过降低糖酵解酶活性来抑制克隆白血病细胞的生长。

• 发表时间: 2005
• 期刊: Oncology Report 14
• 作者: Harada Y;et al.
• 通讯作者: et al.

Gelatinase expression in ocular surface disorders

明胶酶在眼表疾病中的表达

• 发表时间: 2004
• 期刊: Japanese Journal of Ophthalmology 48
• 作者: Sakimoto T;et al.
• 通讯作者: et al.

Gelatinase expression in ocular surface disorders.

明胶酶在眼表疾病中的表达。

• 发表时间: 2004
• 期刊: Jpn J Ophthalmol 48
• 作者: Sakimoto;T.;Shoji;J.;Kanno;H.;Sawa;M.
• 通讯作者: M.