Ketamine causes mitochondrial dysfunction in human induced pluripotent stem cell-derived neurons.
Ketamine causes mitochondrial dysfunction in human induced pluripotent stem cell-derived neurons.
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DOI：
10.1371/journal.pone.0128445
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Makita K
中科院分区：
文献类型：
--
作者：
Ito H;Uchida T;Makita K
氯胺酮在非人类哺乳动物神经元中的毒性已被证明。为研究氯胺酮对神经细胞的毒性作用，建立了体外培养的人诱导多能干细胞(IPSCs)神经元模型，并对其毒性机制进行了初步探讨。用0、2 0、10 0和5 0 0μM氯胺酮处理人IPSC来源的多巴胺能神经元6和2 4h，通过半胱氨酸天冬氨酸氨基转移酶3/7活性、活性氧产生、线粒体膜电位、三磷酸腺苷浓度、神经递质重摄取活性和NADH/NAD+比值来评价氯胺酮的毒性。用透射电子显微镜和共聚焦显微镜分析线粒体的形态变化。与未处理细胞相比，IPSC来源的神经元暴露于500μM氯胺酮24小时后，caspase3/7活性增加40%(P<；0.01)，ROS产生增加14%(P<；0.01)，线粒体膜电位降低81%(P<；0.01)。低浓度氯胺酮(100μM)不激活半胱氨酸半胱氨酸氨基转移酶(Caspase)，可降低心肌细胞三磷酸腺苷水平(22%，P<0.01)，增加NADH/NAD+比值(46%，P<0.05)。透射电子显微镜显示，在100μM氯胺酮浓度下，线粒体分裂和自噬作用增强，提示线粒体功能障碍先于ROS的产生和caspase的激活。我们建立了一种评估氯胺酮对IPSC来源神经元的神经毒性的体外模型。目前的数据表明，最初的线粒体功能障碍和自噬可能与其对线粒体电子传递系统的抑制作用有关，这是氯胺酮神经毒性的基础。较高浓度的氯胺酮可诱导人神经元产生ROS和诱导细胞凋亡。
Ketamine toxicity has been demonstrated in nonhuman mammalian neurons. To study the toxic effect of ketamine on human neurons, an experimental model of cultured neurons from human induced pluripotent stem cells (iPSCs) was examined, and the mechanism of its toxicity was investigated. Human iPSC-derived dopaminergic neurons were treated with 0, 20, 100 or 500 μM ketamine for 6 and 24 h. Ketamine toxicity was evaluated by quantification of caspase 3/7 activity, reactive oxygen species (ROS) production, mitochondrial membrane potential, ATP concentration, neurotransmitter reuptake activity and NADH/NAD+ ratio. Mitochondrial morphological change was analyzed by transmission electron microscopy and confocal microscopy. Twenty-four-hour exposure of iPSC-derived neurons to 500 μM ketamine resulted in a 40% increase in caspase 3/7 activity (P < 0.01), 14% increase in ROS production (P < 0.01), and 81% reduction in mitochondrial membrane potential (P < 0.01), compared with untreated cells. Lower concentration of ketamine (100 μM) decreased the ATP level (22%, P < 0.01) and increased the NADH/NAD+ ratio (46%, P < 0.05) without caspase activation. Transmission electron microscopy showed enhanced mitochondrial fission and autophagocytosis at the 100 μM ketamine concentration, which suggests that mitochondrial dysfunction preceded ROS generation and caspase activation. We established an in vitro model for assessing the neurotoxicity of ketamine in iPSC-derived neurons. The present data indicate that the initial mitochondrial dysfunction and autophagy may be related to its inhibitory effect on the mitochondrial electron transport system, which underlies ketamine-induced neural toxicity. Higher ketamine concentration can induce ROS generation and apoptosis in human neurons.
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AMP激活的蛋白激酶对ULK1（HATG1）的磷酸化将能量传感连接到线粒体。
DOI：
10.1126/science.1196371
发表时间：
2011-01-28
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Egan DF;Shackelford DB;Mihaylova MM;Gelino S;Kohnz RA;Mair W;Vasquez DS;Joshi A;Gwinn DM;Taylor R;Asara JM;Fitzpatrick J;Dillin A;Viollet B;Kundu M;Hansen M;Shaw RJ
通讯作者：
Shaw RJ
影响因子：
3.4
作者：
Mouli, Pradeep K.;Twig, Gilad;Shirihai, Orian S.
通讯作者：
Shirihai, Orian S.
影响因子：
4.1
作者：
ALDRIDGE, WN;PARKER, VH
通讯作者：
PARKER, VH
影响因子：
9.8
作者：
Lee, S. -T.;Wu, T. -T.;Chen, R. -M.
通讯作者：
Chen, R. -M.
DOI：
10.1083/jcb.200412022
发表时间：
2005-05-09
期刊：
The Journal of cell biology
影响因子：
--
作者：
Komatsu M;Waguri S;Ueno T;Iwata J;Murata S;Tanida I;Ezaki J;Mizushima N;Ohsumi Y;Uchiyama Y;Kominami E;Tanaka K;Chiba T
通讯作者：
Chiba T