Pink1 attenuates propofol-induced apoptosis and oxidative stress in developing neurons
Pink1 attenuates propofol-induced apoptosis and oxidative stress in developing neurons
复制标题
Pink1 减弱异丙酚诱导的神经元凋亡和氧化应激
DOI：
10.1007/s00540-017-2431-2
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发表时间：
2018-02-01
影响因子：
2.8
通讯作者：
Xue, Zhanggang
中科院分区：
文献类型：
--
作者：
Liang, Chao;Du, Fang;Xue, Zhanggang
背景丙泊酚对发育中的神经元的毒性作用机制至今尚未完全阐明。我们研究了PTEN诱导的激酶1（Pink 1），一种抗氧化蛋白，在丙泊酚诱导的发育中neurons.Materials和MethodsPrimary海马神经元细胞凋亡中的作用，分离自新生Sprague-Dawley大鼠，丙泊酚20 μM，作用2，4，6和12 h。随后，神经元经历Pink 1的过表达和敲低，然后暴露于丙泊酚（20 μM，6 h）。TUNEL法检测神经细胞凋亡。用2，7-二氯二氢荧光素二乙酸酯探针检测神经元中活性氧（ROS）的产生，用Western blotting或real-time polymerase chain reaction.ResultsPropofol处理时间依赖性地增加TUNEL阳性神经元的数量和切割的caspase-3和B细胞淋巴瘤2（BcL-2）相关X蛋白的表达水平，但BcL-2的表达水平降低。异丙酚处理后，Pink 1 mRNA和蛋白表达水平呈时间依赖性降低。丙泊酚处理增加了ROS的产生和氧化应激的标志物2，4-二硝基苯酚和4-羟基壬烯醛。结论Pink 1在异丙酚诱导的神经元凋亡中起重要作用。我们的研究结果可能提供一些新的见解丙泊酚诱导的神经毒性发育中的神经元。
BackgroundThe underlying mechanisms of propofol-induced neurotoxicity in developing neurons are still not completely understood. We examined the role of PTEN-induced kinase 1 (Pink1), an antioxidant protein, in propofol-induced apoptosis in developing neurons.Materials and methodsPrimary hippocampal neurons isolated from neonatal Sprague–Dawley rats were exposed to propofol 20 μM for 2, 4, 6 and 12 h. Subsequently, neurons underwent overexpression and knockdown of Pink1, followed by propofol exposure (20 μM, 6 h). Neuron apoptosis was detected by terminal transferase deoxyuridine triphosphate-biotin nick-end labeling (TUNEL). Reactive oxygen species (ROS) production in neurons was detected by using a 2,7-dichlorodihydro-fluorescein diacetate probe and target protein or mRNA levels were analyzed by Western blotting or real-time polymerase chain reaction.ResultsPropofol treatment time-dependently increased the number of TUNEL-positive neurons and the expression levels of cleaved caspase-3 and B-cell lymphoma 2 (BcL-2) associated X protein, but decreased expression levels of BcL-2. Furthermore, propofol treatment time-dependently reduced the expression levels of Pink1 mRNA and protein. ROS production and the markers of oxidative stress, 2,4-dinitrophenol and 4-hydroxynonenal, were increased by propofol treatment. However, these propofol-induced changes were significantly restored by Pink1 overexpression.ConclusionsPink1 plays an important role in neuronal apoptosis induced by propofol. Our results may provide some new insights in propofol-induced neurotoxicity in developing neurons.