Altered Mitochondrial Dynamics Contributes to Propofol-induced Cell Death in Human Stem Cell-derived Neurons.
Altered Mitochondrial Dynamics Contributes to Propofol-induced Cell Death in Human Stem Cell-derived Neurons.
复制标题
线粒体动力学的改变会导致丙泊酚诱导的人类干细胞衍生神经元的细胞死亡。
DOI：
10.1097/aln.0000000000000857
复制
发表时间：
2015-11
期刊：
影响因子：
8.8
通讯作者：
Bai X
中科院分区：
文献类型：
--
作者：
Twaroski DM;Yan Y;Zaja I;Clark E;Bosnjak ZJ;Bai X
对发育中动物的研究表明，当麻醉剂在生命早期使用时，会导致神经细胞死亡和学习障碍。人胚胎干细胞（hESC）衍生神经元的发育为了解麻醉剂对发育中的人类神经元的影响提供了有价值的工具。不平衡的线粒体融合/分裂导致各种病理状况，包括神经变性。本研究的目的是剖析线粒体动力学在丙泊酚诱导的神经毒性中的作用。TUNEL染色用于评估hESC衍生的神经元中的细胞死亡。使用TOM 20染色和电子显微镜评估线粒体分裂。通过Western blot评估线粒体分裂相关蛋白的表达，并使用共聚焦显微镜评估线粒体通透性转换孔（mPTP）的开放时间。暴露于20 μg/mL丙泊酚6小时后，细胞死亡率从对照组的3.18±0.17%增加至9.6±0.95%，并导致线粒体分裂有害增加（n=5个盖玻片/组），伴随着激活的动力蛋白相关蛋白1（Drp 1）和细胞周期蛋白依赖性激酶1（CDK 1）（负责线粒体分裂的关键蛋白）表达增加。丙泊酚暴露也诱导mPTP的开放时间从对照治疗组的118.9±3.1秒提前至73.3±1.6秒。用线粒体分裂阻断剂mdivi-1预处理细胞可挽救丙泊酚诱导的毒性、线粒体分裂和mPTP开放时间（n=75个细胞/组）。抑制CDK 1减弱了细胞死亡和分裂的增加以及活化的Drp 1表达的增加。这些数据首次证明丙泊酚诱导的神经毒性通过线粒体分裂/mPTP介导的途径发生。
Studies in developing animals have shown that when anesthetic agents are administered early in life, it can lead to neuronal cell death and learning disabilities. Development of human embryonic stem cell (hESC)-derived neurons has provided a valuable tool for understanding the effects of anesthetics on developing human neurons. Unbalanced mitochondrial fusion/fission leads to various pathological conditions including neurodegeneration. The aim of this study was to dissect the role of mitochondrial dynamics in propofol-induced neurotoxicity. TUNEL staining was used to assess cell death in hESC-derived neurons. Mitochondrial fission was assessed using TOM20 staining and electron microscopy. Expression of mitochondrial fission-related proteins was assessed by Western blot and confocal microscopy was used to assess opening time of the mitochondrial permeability transition pore (mPTP). Exposure to 6 hours of 20 μg/mL propofol increased cell death from 3.18±0.17% in the control-treated group to 9.6±0.95% and led to detrimental increases in mitochondrial fission (n=5 coverslips/group) accompanied by increased expression of activated dynamin-related protein 1 (Drp1) and cyclin-dependent kinase 1 (CDK1), key proteins responsible for mitochondrial fission. Propofol exposure also induced earlier opening of the mPTP from 118.9±3.1 seconds in the control-treated group to 73.3±1.6 seconds. Pretreatment of the cells with mdivi-1, a mitochondrial fission blocker rescued the propofol-induced toxicity, mitochondrial fission and mPTP opening time (n=75 cells/group). Inhibiting CDK1 attenuated the increase in cell death and fission and the increase in expression of activated Drp1. These data demonstrate for the first time that propofol-induced neurotoxicity occurs through a mitochondrial fission/mPTP-mediated pathway.