Ozone alleviates ischemia/reperfusion injury by inhibiting mitochondrion-mediated apoptosis pathway in SH-SY5Y cells
Ozone alleviates ischemia/reperfusion injury by inhibiting mitochondrion-mediated apoptosis pathway in SH-SY5Y cells
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DOI：
10.1002/cbin.11294
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发表时间：
2020-01-21
影响因子：
3.9
通讯作者：
Zhu, Yi-Min
中科院分区：
文献类型：
--
作者：
Cai, Hua-An;Tao, Xi;Zhu, Yi-Min
脑缺血/再灌注（I/R）损伤很常见，通常会引起严重的并发症。臭氧已用于保护包括小脑在内的多个器官的动物模型保护I/R损伤，但详细机制尚不清楚。 3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四唑溴化物（MTT）测定法和乳酸脱氢酶测量，以确定臭氧对细胞活性和SH-SY5Y细胞损伤的影响。通过酶联免疫吸附测定法测量了一些氧化还原项目，例如过氧化氢酶（CAT），丙二醛（MDA），谷胱甘肽过氧化物酶（GSH-PX）和超氧化物歧化酶（SOD）。线粒体膜电位（DELTA PSI（M））通过JC-1测定确定。通过蛋白质印迹测量细胞质和线粒体中的细胞色素-C（CYT-C）水平。通过流式细胞术确定凋亡，并通过定量的实时聚合酶链反应和蛋白质印迹检测到一些与凋亡相关的分子。臭氧通过增加GSH-PX，SOD，CAT和减少MDA来减轻氧化损伤。臭氧减少了由I/R损伤引起的线粒体损伤，并抑制了SH-SY5Y细胞中Cyt-C从线粒体到细胞质的释放。 I/R引起的细胞凋亡受到臭氧的抑制，臭氧可以通过增加Bcl-2/Bax的比率并抑制SH-SY5Y细胞中的caspase信号通路来降低凋亡。臭氧具有维持氧化还原稳态，减少线粒体损伤并抑制I/R诱导的神经细胞凋亡的能力。因此，臭氧可能是针对脑I/R损伤的有前途的保护策略。
Cerebral ischemia/reperfusion (I/R) injuries are common and often cause severe complications. Ozone has been applied for protecting I/R injury in animal models of several organs including cerebra, but the detailed mechanism remains unclear. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lactate dehydrogenase measurement were used to determine the influence of ozone on cell activity and damage of SH-SY5Y cells. Some redox items such as catalase (CAT), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) were measured by enzyme-linked immunosorbent assay. The mitochondrial membrane potential (Delta psi(m)) was determined by JC-1 assay. Cytochrome-c (cyt-c) level in the cytoplasm and mitochondrion was measured by western blotting. Apoptosis was determined by flow cytometry, and some apoptosis-related molecules were detected by quantitative real-time polymerase chain reaction and western blotting. Ozone alleviated oxidative damage by increasing GSH-Px, SOD, CAT, and decreasing MDA. Ozone decreased mitochondrial damage caused by I/R injury and inhibited the release of cyt-c from mitochondrion to cytoplasm in SH-SY5Y cells. The cell apoptosis caused by I/R was inhibited by ozone, and ozone could decrease apoptosis by increasing the ratio of Bcl-2/Bax and inhibiting caspase signaling pathway in SH-SY5Y cells. Ozone has the ability of maintaining redox homeostasis, decreasing mitochondrion damage, and inhibiting neurocytes apoptosis induced by I/R. Therefore, ozone may be a promising protective strategy against cerebral I/R injury.