High Glucose Attenuates Anesthetic Cardioprotection in Stem-Cell-Derived Cardiomyocytes: The Role of Reactive Oxygen Species and Mitochondrial Fission.
High Glucose Attenuates Anesthetic Cardioprotection in Stem-Cell-Derived Cardiomyocytes: The Role of Reactive Oxygen Species and Mitochondrial Fission.
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DOI：
10.1213/ane.0000000000001254
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发表时间：
2016-05
影响因子：
5.7
通讯作者：
Bosnjak ZJ
中科院分区：
文献类型：
--
作者：
Canfield SG;Zaja I;Godshaw B;Twaroski D;Bai X;Bosnjak ZJ
高血压可减弱异氟醚在缺血再灌注损伤中的心脏保护作用。以往的研究表明，活性氧（ROS）和线粒体分裂增加在缺血再灌注损伤心肌细胞死亡中发挥作用。为了研究葡萄糖浓度在缺血-再灌注（有和没有麻醉保护）期间ROS产生和线粒体分裂中的作用，我们使用了人诱导的多能干细胞衍生的心肌细胞（iPSC-CM）的新平台。使用免疫组织化学和测量收缩性，通过CM特异性标志物的表达来表征来自iPSC的心肌细胞分化。将iPSC-CM暴露于不同的葡萄糖条件（5、11、25 mM）24小时。线粒体渗透性转换孔（mPTP）开放，细胞活力，和ROS产生的终点被用来评估各种治疗条件的影响。通过使用共聚焦显微镜观察破碎的线粒体来监测线粒体分裂。活化动力蛋白相关蛋白1（Drp 1）的表达，负责线粒体分裂的关键蛋白质进行了评估。成功地从iPSC分化出心肌细胞。升高的葡萄糖条件（11和25 mM）显著增加ROS产生，而仅25 mM高葡萄糖条件诱导线粒体分裂并增加iPSC-CM中活化的Drp 1的表达。异氟烷延迟mPTP开放并保护iPSC-CM在5和11 mM葡萄糖条件下免受氧化应激，达到与先前在各种分离的动物心肌细胞中观察到的相似水平。在25 mM葡萄糖条件下，用Trolox清除ROS或用mdivi-1抑制线粒体分裂恢复了iPSC-CM中的麻醉心脏保护作用。人iPSC-CM是用于研究异氟烷心脏保护的有用的相关模型，并且可以被操纵以重现复杂的临床扰动。我们证明，异氟烷在高血糖条件下的心脏保护作用可以通过清除ROS或抑制线粒体分裂来恢复。这些发现可能有助于进一步了解和指导恢复高血糖患者的药物心脏保护。
Hyperglycemia can blunt the cardioprotective effects of isoflurane in the setting of ischemia-reperfusion injury. Previous studies suggest that reactive oxygen species (ROS) and increased mitochondrial fission play a role in cardiomyocyte death during ischemia-reperfusion injury. To investigate, the role of glucose concentration in ROS production and mitochondrial fission during ischemia-reperfusion (with and without anesthetic protection), we used the novel platform of human-induced pluripotent stem-cell-derived cardiomyocytes (iPSC-CMs). Cardiomyocyte differentiation from iPSC was characterized by the expression of CM-specific markers using immunohistochemistry and by measuring contractility. iPSC-CMs were exposed to varying glucose conditions (5, 11, 25 mM) for 24 hours. Mitochondrial permeability transition pore (mPTP) opening, cell viability, and ROS generation end-points were used to assess the effects of various treatment conditions. Mitochondrial fission was monitored by the visualization of fragmented mitochondria using confocal microscopy. Expression of activated dynamin-related protein 1 (Drp1), a key protein responsible for mictochodrial fission was assessed by western blot. Cardiomyocytes were successfully differentiated from iPSC. Elevated glucose conditions (11 and 25 mM) significantly increased ROS generation, while only the 25 mM high glucose condition induced mitochondrial fission and increased the expression of activated Drp1 in iPSC-CMs. Isoflurane delayed mPTP opening and protected iPSC-CMs from oxidative stress in 5 and 11 mM glucose conditions to a similar level as previously observed in various isolated animal cardiomyocytes. Scavenging ROS with Trolox or inhibiting mitochondrial fission with mdivi-1 restored the anesthetic cardioprotective effects in iPSC-CMs in 25 mM glucose conditions. Human iPSC-CM is a useful, relevant model for studying isoflurane cardioprotection, and can be manipulated to recapitulate complex clinical perturbations. We demonstrate that the cardioprotective effects of isoflurane in elevated glucose conditions can be restored by scavenging ROS or inhibiting mitochondrial fission. These findings may contribute to further understanding and guidance for restoring pharmacological cardioprotection in hyperglycemic patients.