Department of Anesthesiology, Fushun Mining Bureau General Hospital of Liaoning Health Industry Group, Fushun, Liaoning, China.
Department of Anesthesiology, Fushun Second People's Hospital, Fushun, Liaoning, China.
Hum Exp Toxicol. 2021 Jul;40(7):1163-1172. doi: 10.1177/0960327121989416. Epub 2021 Jan 29.
To investigate how nuclear factor-E2-related factor 2 (Nrf2) involved in the protective effect of isoflurane (Iso) preconditioning in oxygen glucose deprivation (OGD)-induced cortical neuron injury.
Primary mouse cortical neurons were divided into Control, ML385 (an Nrf2 inhibitor), Iso, Iso + ML385, OGD, ML385 + OGD, Iso + OGD, and Iso + ML385 + OGD groups. Lactate dehydrogenase activity (LDH) release and oxidative stress indexes were quantified. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect cell viability, Annexin V-FITC/propidium iodide (PI) staining to measure cell apoptosis, dichloro-dihydro-fluorescein diacetate (DCFH-DA) method to test reactive oxygen species (ROS), and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and Western blotting to evaluate genes and protein expression.
Iso preconditioning reduced LDH release and inhibited cell cytotoxicity in OGD-induced cortical neurons, which was abolished by ML385. Iso preconditioning increased the Nrf2 nuclear translocation in cortical neurons. Meanwhile, Iso decreased the OGD-induced apoptosis with the down-regulations of Bax and Caspase-3 and the up-regulation of Bcl-2, which was reversed by ML385. OGD enhanced the level of ROS and malondialdehyde (MDA) in cortical neurons, but reduced the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), which were aggravated in ML385 + OGD group and mitigated in Iso + OGD group. No observable difference was found between OGD group and Iso + ML385 + OGD group regarding apoptosis-related proteins and oxidative stress-related indexes.
Iso preconditioning up-regulated Nrf2 level to play its protective role in OGD-induced mouse cortical neuron injury.
研究核因子-2 相关因子 2（Nrf2）在异氟醚（Iso）预处理对氧葡萄糖剥夺（OGD）诱导的皮质神经元损伤中的保护作用中的作用机制。
将原代培养的小鼠皮质神经元分为对照组、ML385（Nrf2 抑制剂）组、Iso 组、Iso+ML385 组、OGD 组、ML385+OGD 组、Iso+OGD 组和 Iso+ML385+OGD 组。定量检测乳酸脱氢酶（LDH）释放和氧化应激指标。噻唑蓝（MTT）比色法检测细胞活力，Annexin V-FITC/碘化丙啶（PI）染色法检测细胞凋亡，二氯二氢荧光素二乙酸酯（DCFH-DA）法检测活性氧（ROS），实时荧光定量聚合酶链反应（qRT-PCR）和 Western blot 法检测基因和蛋白表达。
Iso 预处理可减少 OGD 诱导的皮质神经元中 LDH 的释放和抑制细胞毒性，而 ML385 则可消除 Iso 的这种作用。Iso 预处理可增加皮质神经元中 Nrf2 的核转位。同时，Iso 可降低 Bax 和 Caspase-3 的表达，上调 Bcl-2 的表达，从而减少 OGD 诱导的皮质神经元凋亡，而 ML385 则可逆转 Iso 的这种作用。OGD 可增加皮质神经元中 ROS 和丙二醛（MDA）的水平，但降低超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）的活性，而在 ML385+OGD 组中这些指标加重，在 Iso+OGD 组中则减轻。在凋亡相关蛋白和氧化应激相关指标方面，OGD 组与 Iso+ML385+OGD 组之间没有观察到明显差异。
Iso 预处理上调 Nrf2 水平，在 OGD 诱导的小鼠皮质神经元损伤中发挥其保护作用。
