Marked hyperglycemia attenuates anesthetic preconditioning in human-induced pluripotent stem cell-derived cardiomyocytes.
Marked hyperglycemia attenuates anesthetic preconditioning in human-induced pluripotent stem cell-derived cardiomyocytes.
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DOI：
10.1097/aln.0b013e3182655e96
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发表时间：
2012-10
期刊：
影响因子：
8.8
通讯作者：
Bosnjak ZJ
中科院分区：
文献类型：
--
作者：
Canfield SG;Sepac A;Sedlic F;Muravyeva MY;Bai X;Bosnjak ZJ
麻醉预处理可保护心肌细胞免受氧化应激损伤，但对糖尿病患者无效。为了解决高血糖症在糖尿病个体无法预处理中的作用，我们使用从患有或不患有2型糖尿病的患者产生的诱导多能干细胞分化的人心肌细胞（分别为DM-iPSC-和N-iPSC-CM）来研究预处理在不同葡萄糖条件（5、11和25 mM）下的功效。诱导多能干细胞通过定向分化诱导产生心肌细胞。在随后的研究中，心肌细胞通过用心脏特异性启动子驱动的增强型绿色荧光蛋白进行遗传标记来鉴定。通过乳酸脱氢酶测定分析细胞活力。共聚焦显微镜观察线粒体通透性转换孔和线粒体腺苷三磷酸敏感性钾通道的开放情况。异氟烷（0.5 mM）预处理保护N-iPSC-和DM-iPSC-CM在5 mM和11 mM葡萄糖中免受氧化应激诱导的乳酸脱氢酶释放和线粒体通透性转换孔开放。异氟烷在5 mM和11 mM葡萄糖中的N-iPSC-CM和5 mM葡萄糖中的DM-iPSC-CM中触发线粒体腺苷-5 ′-三磷酸敏感性钾通道开放。25 mM葡萄糖破坏DM-iPSC-和N-iPSC-CM中的麻醉预处理介导的保护。在11 mM和25 mM葡萄糖中的DM-iPSC-CM和在25 mM葡萄糖中的N-iPSC-CM中，线粒体腺苷-5 ′-三磷酸敏感性钾通道的开放被破坏。来自健康供体和患有特定疾病（如本研究中的糖尿病）的患者的心肌细胞为在人类相关模型中研究基因型和表型相关病理提供了可能性。
Anesthetic preconditioning protects cardiomyocytes from oxidative stress-induced injury, but it is ineffective in patients with diabetes mellitus. To address the role of hyperglycemia in the inability of diabetic individuals to be preconditioned, we used human cardiomyocytes differentiated from induced pluripotent stem cells generated from patients with or without type 2 diabetes mellitus (DM-iPSC- and N-iPSC-CMs, respectively) to investigate the efficacy of preconditioning in varying glucose conditions (5, 11, and 25 mM). Induced pluripotent stem cells were induced to generate cardiomyocytes by directed differentiation. For subsequent studies, cardiomyocytes were identified by genetic labeling with enhanced green fluorescent protein driven by a cardiac-specific promoter. Cell viability was analyzed by lactate dehydrogenase assay. Confocal microscopy was utilized to measure opening of the mitochondrial permeability transition pore and the mitochondrial adenosine-5′-triphosphate-sensitive potassium channels. Isoflurane (0.5 mM) preconditioning protected N-iPSC- and DM-iPSC-CMs from oxidative stress-induced lactate dehydrogenase release and mitochondrial permeability transition pore opening in 5 mM and 11 mM glucose. Isoflurane triggered mitochondrial adenosine-5′-triphosphate-sensitive potassium channel opening in N-iPSC-CMs in 5 mM and 11 mM glucose and in DM-iPSC-CMs in 5 mM glucose. 25 mM glucose disrupts anesthetic preconditioning-mediated protection in DM-iPSC- and N-iPSC-CMs. The opening of mitochondrial adenosine-5′-triphosphate-sensitive potassium channels are disrupted in DM-iPSC-CMs in 11 mM and 25 mM glucose and in N-iPSC-CMs in 25 mM glucose. Cardiomyocytes derived from healthy donors and patients with a specific disease, such as diabetes in this study, open possibilities in studying genotype and phenotype related pathologies in a human relevant model.