Liang Suixin, Ti Yunxing, Li Xiuhong, Zhou Wenjia
Department of CICU, Shenzhen Children's Hospital, Shenzhen City, Guangdong Province, People's Republic of China.
Department of Cardiothoracic Surgery, Shenzhen Children's Hospital, Shenzhen City, Guangdong Province, People's Republic of China.
Neuropsychiatr Dis Treat. 2023 Jul 17;19:1625-1631. doi: 10.2147/NDT.S412227. eCollection 2023.
Moderate therapeutic hypothermia is protective against several cellular stressors. However, the mechanisms behind this protection are not entirely known. In the current investigation, we investigated that therapeutic hypothermia at 33°C administered following peroxide-induced oxidative stress might protect human oligodendroglioma cells using an in vitro model.
Tert-butyl peroxide treatment for one hour significantly increased cell apoptosis and suppressed cell viability. In the range of 50-1000 M tert-butyl peroxide, this cell death was dose-dependent. MTT assay and cell apoptosis assay were applied to analyze cell viability/death at 24 hours after peroxide-induced stress. Therapeutic hypothermia at 33°C delivered for two hours after peroxide exposure significantly increased cell viability and suppressed cell death. Even 15 minutes after peroxide washout when delayed hypothermia was used, this protection was still apparent. Three FDA-approved antioxidants (Tempol, EUK134, and Edaravone at 100 M) were added immediately after tert-butyl peroxide, followed by hypothermia treatment. These three antioxidants greatly increased cell viability and cell apoptosis. RT-qPCR was applied to determine the effects of hypothermia treatment on the expression of caspase-3 and -8 as well as tumor necrosis factor-alpha (TNF-α). Therapeutic hypothermia significantly downregulated these three factors.
Overall, these findings confirmed that hypothermia and antioxidants quenching reactive oxygen species may lower mitochondrial oxidative stress and/or apoptotic pathways. Further investigation are needed to investigate the role of hypothermia in other cell models.
适度治疗性低温对多种细胞应激源具有保护作用。然而，这种保护背后的机制尚不完全清楚。在本研究中，我们使用体外模型研究了在过氧化氢诱导的氧化应激后给予33°C的治疗性低温是否能保护人少突胶质细胞瘤细胞。
用叔丁基过氧化氢处理1小时可显著增加细胞凋亡并抑制细胞活力。在50 - 1000μM叔丁基过氧化氢范围内，这种细胞死亡呈剂量依赖性。用过氧化氢诱导应激24小时后，采用MTT法和细胞凋亡检测法分析细胞活力/死亡情况。在过氧化氢暴露后给予2小时33°C的治疗性低温可显著提高细胞活力并抑制细胞死亡。即使在使用延迟低温的过氧化氢洗脱15分钟后，这种保护作用仍然明显。在叔丁基过氧化氢处理后立即加入三种美国食品药品监督管理局（FDA）批准的抗氧化剂（Tempol、EUK134和100μM依达拉奉），然后进行低温治疗。这三种抗氧化剂可显著提高细胞活力并抑制细胞凋亡。采用RT-qPCR法检测低温治疗对caspase-3、-8以及肿瘤坏死因子-α（TNF-α）表达的影响。治疗性低温可显著下调这三种因子的表达。
总体而言，这些发现证实低温和抗氧化剂清除活性氧可能会降低线粒体氧化应激和/或凋亡途径。需要进一步研究以探讨低温在其他细胞模型中的作用。
