Sevoflurane is commonly used in clinical anesthesia. However, some reports indicated that Sevoflurane could induce mitochondrial injury and neuroapoptosis. Although the mechanism remains unclear, evidence points to the increase of intracellular calcium after administration of Sevoflurane. Herein, we sought whether the increment of intracellular Ca2+ caused by Sevoflurane administration could induce mitochondrial injury and apoptosis in primary neurons of the hippocampus. Fluo-4-acetoxymethyl ester Ca2+ probe was used for measuring intracellular Ca2+ concentrations. LDH assay, CCK-8 assay, and Western blotting were performed to confirm Sevoflurane-induced neuroapoptosis. ROS, mPTP, and ATP production were assayed to reveal mitochondrial injury. Our results indicated that Sevoflurane increased intracellular Ca2+ and neuronal death. Sevoflurane also elevated ROS and the opening of mPTP, and decreased ATP production in neurons. The expression of cytochrome c, cleaved caspase-9, cleaved caspase-3, and the ratio of Bax/Bcl-2 were also increased. By using calcium channel blocker Nimodipine, the increase of intracellular Ca2+ was attenuated, and the death rate of neurons, the ROS and opening of mPTP, decreased ATP production, the expressions of cytochrome c, cleaved caspase-9, cleaved caspase-3 and the ratio of Bax/Bcl-2 were alleviated. Our study suggested that Sevoflurane could increase intracellular Ca2+ to induce mitochondrial injury and mitochondria-mediated neuroapoptosis in neurons.

中文翻译：

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七氟烷增加细胞内钙离子诱导线粒体损伤和神经细胞凋亡

七氟烷常用于临床麻醉。然而，一些报道表明，七氟烷可诱导线粒体损伤和神经细胞凋亡。虽然机制尚不清楚，但有证据表明施用七氟醚后细胞内钙增加。在此，我们探索七氟醚给药引起的细胞内Ca2+的增加是否能诱导海马原代神经元的线粒体损伤和凋亡。Fluo-4-乙酰氧基甲酯Ca2+探针用于测量细胞内Ca2+浓度。进行 LDH 测定、CCK-8 测定和蛋白质印迹以确认七氟烷诱导的神经细胞凋亡。测定 ROS、mPTP 和 ATP 产生以揭示线粒体损伤。我们的结果表明，七氟烷增加了细胞内 Ca2+ 和神经元死亡。七氟烷还升高了 ROS 和 mPTP 的开放，并降低了神经元中 ATP 的产生。细胞色素 c、cleaved caspase-9、cleaved caspase-3 和 Bax/Bcl-2 的表达也增加。使用钙通道阻滞剂尼莫地平后，细胞内Ca2+的增加减弱，神经元的死亡率、ROS和mPTP的开放、ATP的产生、细胞色素c、cleaved caspase-9、cleaved caspase-3和Bax/Bcl-2 的比率得到缓解。我们的研究表明，七氟烷可以增加细胞内 Ca2+ 以诱导线粒体损伤和线粒体介导的神经元神经细胞凋亡。使用钙通道阻滞剂尼莫地平后，细胞内Ca2+的增加减弱，神经元的死亡率、ROS和mPTP的开放、ATP的产生、细胞色素c、cleaved caspase-9、cleaved caspase-3和Bax/Bcl-2 的比率得到缓解。我们的研究表明，七氟烷可以增加细胞内 Ca2+ 以诱导线粒体损伤和线粒体介导的神经元神经细胞凋亡。使用钙通道阻滞剂尼莫地平后，细胞内Ca2+的增加减弱，神经元的死亡率、ROS和mPTP的开放、ATP的产生、细胞色素c、cleaved caspase-9、cleaved caspase-3和Bax/Bcl-2 的比率得到缓解。我们的研究表明，七氟烷可以增加细胞内 Ca2+ 以诱导线粒体损伤和线粒体介导的神经元神经细胞凋亡。