p53 MEDIATES AUTOPHAGY ACTIVATION AND MITOCHONDRIA DYSFUNCTION IN KAINIC ACID-INDUCED EXCITOTOXICITY IN PRIMARY STRIATAL NEURONS
p53 MEDIATES AUTOPHAGY ACTIVATION AND MITOCHONDRIA DYSFUNCTION IN KAINIC ACID-INDUCED EXCITOTOXICITY IN PRIMARY STRIATAL NEURONS
复制标题
p53 介导红藻氨酸诱导的初级纹状体神经元兴奋性毒性中的自噬激活和线粒体功能障碍
DOI：
10.1016/j.neuroscience.2012.01.018
复制
发表时间：
2012-04-05
期刊：
影响因子：
3.3
通讯作者：
Wang, Y.
中科院分区：
文献类型：
--
作者：
Dong, X. X.;Wang, Y. R.;Wang, Y.
本研究旨在探讨 p53 是否介导红藻氨酸 (KA) 诱导的兴奋性毒性中原代纹状体神经元的自噬激活和线粒体功能障碍。采用KA建立原代纹状体神经元兴奋毒性模型。通过蛋白质印迹和免疫染色检查 p53、微管相关蛋白 1 轻链 3 (LC3)、Beclin1 和 p62 的水平。还用电子显微镜测定自噬激活。为了评估 KA 诱导的兴奋性毒性中 p53 对自噬激活和线粒体功能障碍的贡献，使用 p53 抑制剂 Pifithrin-α 预处理后，LC3、Beclin1 和 p62 的蛋白水平、线粒体跨膜电位和线粒体活性氧 (ROS) (PFT-α) 和自噬抑制剂 3-甲基腺嘌呤 (3-MA) 进行了分析。 KA 治疗后会诱发兴奋性毒性神经元损伤，如乳酸脱氢酶 (LDH) 渗漏增加所证明，并且 PFT-α 显着抑制该损伤。 Western blot和免疫染色显示p53蛋白的诱导发生在细胞质和细胞核中。 KA 处理后观察到自噬蛋白 LC3 和 Beclin1 增加，而 p62 蛋白水平下降。电镜分析显示细胞质中自噬体增加。 LC3、Beclin1 和 p62 水平的变化被 PFT-alpha、PFT-mu、3-MA 和 E64d 阻断，但 Z-DEVD-FMK 不能阻断。 JC-1染色显示兴奋性毒性损伤后线粒体膜电位去极化。 Mito-tracker 和 RedoxSensor Red CC-1 染色显示，兴奋性毒性损伤后线粒体 ROS 的产生增加。用 PFT-α 和 3-MA 预处理后，这些效应得到显着抑制。这项研究表明 p53 介导 KA 诱导的纹状体神经元自噬激活和线粒体功能障碍。 (C) 2012 国际广播组织。由爱思唯尔有限公司出版。保留所有权利。
The present study sought to investigate if p53 mediates autophagy activation and mitochondria dysfunction in primary striatal neurons in kainic acid (KA)-induced excitotoxicity. The excitotoxic model of primary striatal neurons was established with KA. The levels of p53, microtubule-associated protein 1 light chain 3 (LC3), Beclin1, and p62 were examined by Western blot and immunostaining. Autophagy activation was also determined with electron microscope. To evaluate the contribution of p53 to autophagy activation and mitochondria dysfunction in KA-induced excitotoxicity, the protein levels of LC3, Beclin1, and p62, the mitochondrial transmembrane potential and the mitochondrial Reactive oxygen species (ROS) after pretreatment with the p53 inhibitor pifithrin-alpha (PFT-alpha) and the autophagy inhibitor 3-methyladenine (3-MA) were analyzed. Excitotoxic neuronal injury was induced after KA treatment as demonstrated by increases in lactate dehydrogenase (LDH) leakage and was significantly inhibited by PFT-alpha. Western blot and immunostaining showed that the induction of p53 protein occurred in the cytosol and the nucleus. Increases in autophagic proteins LC3 and Beclin1 were observed, whereas the protein levels of p62 decreased after KA treatment. Electron microscope analysis showed increased autophagosomes in the cytoplasm. The changes in LC3, Beclin1, and p62 levels were blocked by PFT-alpha, PFT-mu, 3-MA, and E64d but not Z-DEVD-FMK. JC-1 staining showed the depolarization of mitochondrial membrane potential after excitotoxic insult. Mito-tracker and RedoxSensor Red CC-1 staining showed an increased production of mitochondrial ROS after excitotoxic insult. These effects were significantly suppressed after pretreatment with PFT-alpha and 3-MA. This study suggests that p53 mediates KA-induced autophagy activation and mitochondrial dysfunction in striatal neurons. (C) 2012 IBRO. Published by Elsevier Ltd. All rights reserved.