Pathophysiology of mitochondrial control for neuronal death and brain protection

线粒体控制神经元死亡和脑保护的病理生理学

• 批准号: 17591651
• 负责人: IIJIMA Takehiko
• 金额: $ 2.24万
• 依托单位: Kyorin University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17591651/
• 关键词: mitochondria; neuronal death; brain ischemia; membrane potential; intracellular calcium; fluo-3; rhod-2; unipoter; Akt; 細胞内Ca

Ischemic neuronal death is mediated by mitochondria. We have demonstrated that mitochondrial membrane potential showed a specific behavior depending on the grade of ischemic impact. Mitochondrial hyperpolarization has been observed after short-term oxygen-glucose deprivation (OGD) in cultured rat hippocampal neurons. Since hyperpolarization putatively links to subsequent death mode, the role of mitochondrial membrane potential may relate to cytosolic Ca^<++> [Ca^<2+>]c buffering capacity, and signaling pathway of PKB, which regulate apoptotic cascade.1 Cytosolic Ca^<++> [Ca^<2+>]c buffering capacity after OGDHippocampal neurons cultured in dish were anaerobically incubated for 30 or 120 min with glucose-tree medium (30OGD, 120OGD). After OGD, neurons were loaded fluo-3 or rhod-2 for monitoring cytosolic and mitochondrial Ca^<++> concentration ([Ca^<2+>]c, [Ca^<2+>]m). To evaluate mitochondrial membrane potential (MMP), TMRE (tetramethylrhodamie, ethyl ester, perchloratye (TMRE)) was lo … More aded. Neurons were exposed to 0.5mM glutamate to induce depolarization.The [Ca^<2+>]c, was steeply increased by glutamate load and increase in [Ca^<2+>]m followed in control and 30OGD. The [Ca^<2+>]c in 30OGD more rapidly decreased than that in control after it reached peak, and [Ca^<2+>]m progressively increased in 30OGD, while it remained constant in control. The area under the curve of normalized fluorescence of fluo-3 (AUCf) in 30OGD was significantly lower than that oin control, while the area under the curve of normalized fluorescence of rhod-2 (AUCr) in 30OGD was significantly larger than control. In 120OGD, increase in [Ca^<2+>]c and [Ca^<2+>]m was blunted.Mitochondrial Ca buffering capacity after 30OGD was enhanced. This buffering capacity may link to preconditioning after short term ischemic episode.2 OGD and AktWe employed ELISA for detection of phosphorylated Akt. Short-term OGD(30min) increased phosphorylation of Akt, while longer OGD (120min) suppressed phosphorylation. We are now confirming this distinct change by using Western blotting. Less

缺血性神经元死亡由线粒体介导。我们已经证明，线粒体膜电位表现出特定的行为，这取决于缺血影响的程度。在体外培养的大鼠海马神经元上观察到短期氧糖剥夺（OGD）后线粒体超极化现象。由于超极化损伤与随后的死亡方式有关，线粒体膜电位的作用可能与胞浆Ca^++[Ca^2+]c缓冲能力和PKB信号通路有关。1.在培养皿中培养的OGDH诱导神经元后，将细胞溶质Ca^++[Ca^2+]c缓冲能力与葡萄糖一起厌氧孵育30或120分钟。树形培养基（30 OGD、120 OGD）。在OGD后，神经元负载fluo-3或rhod-2以监测胞浆和线粒体Ca^++浓度（[Ca^2+]c，[Ca^2+]m）。用四甲基罗丹明乙酯高氯酸盐（TMRE）测定线粒体膜电位（MMP），并与对照组比较。 ...更多信息 aded.用0.5mM谷氨酸诱导神经元去极化，谷氨酸负荷后[Ca^<2+>]c急剧增加，[Ca^<2+>]m增加，对照组和30 OGD组次之。30 OGD组[Ca^<2+>]c达到峰值后迅速下降，[Ca^<2+>]m逐渐升高，而对照组基本不变。30 OGD组Fluo-3标准化荧光曲线下面积（AUCf）显著低于对照组，而rhod-2标准化荧光曲线下面积（AUCr）显著大于对照组。120 OGD时[Ca^<2+>]c和[Ca^<2+>]m的增加被抑制，30 OGD后线粒体Ca缓冲能力增强。这种缓冲能力可能与短期缺血后的预处理有关。2 OGD和Akt我们采用ELISA检测磷酸化Akt。短期OGD（30 min）增加Akt的磷酸化，而长期OGD（120 min）抑制Akt的磷酸化。我们现在通过使用Western印迹来确认这种明显的变化。少

项目成果

プロポフォールの脳保護作用

异丙酚的脑保护作用

• 发表时间: 2006
• 期刊: Pharmacoanesthesiology 18(1)
• 作者: 飯島 毅彦;巌 康秀
• 通讯作者: 巌 康秀

Neuroprotective effect of propofol on necrosis and apoptosis following oxygen-glucose deprivation - Relationship between mitochondrial membrane potential and mode of death

• DOI: 10.1016/j.brainres.2006.04.117
• 发表时间: 2006-07-12
• 期刊: BRAIN RESEARCH
• 影响因子: 2.9
• 作者: Iijima, Takehiko;Mishima, Tatsuya;Huaa, Yasuhide
• 通讯作者: Huaa, Yasuhide

Neuroprotective effect of propofol. (in Japanese)

异丙酚的神经保护作用。

• 发表时间: 2006
• 期刊: Pharmacoanesthesiology 18(1)
• 作者: Iijima T;Iwao Y.
• 通讯作者: Iwao Y.

Preceding neuroprotective effect of propofol against acute neuronal death is dismissed by subsequent apoptosis following oxygen glucose deprivation in rat cultured hippocampal neurons

在大鼠培养的海马神经元中，氧糖剥夺后的随后的细胞凋亡消除了丙泊酚对急性神经元死亡的先前神经保护作用

• 发表时间: 2006
• 期刊: Brain Research (accepted)
• 作者: Takehiko Iijima;Tatsuya Mishima
• 通讯作者: Tatsuya Mishima