| 郭生龙,朱 洁,谢 瑱,李 鹏,费裕朗,杨 谦.Che-1通过mTOR通路调控自噬在谷氨酸所致神经元损伤中的作用研究[J].,2019,19(18):3456-3460 |
| Che-1通过mTOR通路调控自噬在谷氨酸所致神经元损伤中的作用研究 |
| Study on the Effect of Che-1 on Glutamate-induced Neuronal Injury Via mTOR Regulated Autophagy Pathways |
| 投稿时间：2019-05-10 修订日期：2019-06-04 |
| DOI：10.13241/j.cnki.pmb.2019.18.011 |
| 中文关键词: Che-1 神经元 自噬 神经保护 |
| 英文关键词: Che-1 Neuron Autophagy Neuroprotection |
| 基金项目:国家自然科学基金项目（81601719） |
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| 中文摘要: |
| 摘要 目的：观察谷氨酸（glutamate, Glu）对神经元Che-1蛋白表达的影响，研究过表达Che-1对Glu所致神经元氧化应激性损伤的作用，并以mTOR调控的细胞自噬通路为靶点，探讨Che-1在Glu所致神经元损伤中发挥作用的分子机制。方法：用Glu损伤神经元后，采用免疫学及分子生物学等方法检测Che-1蛋白的表达；用慢病毒转染神经元增加Che-1表达，用乳酸脱氢酶（Lactate dehydrogenase, LDH）释放量和流式细胞术等方法检测神经元凋亡程度，采用免疫荧光染色和免疫印迹法检测神经元自噬关键蛋白表达水平；使用mTOR特异性抑制剂雷帕霉素（Rapamycin）提高神经元自噬水平，并通过检测LDH释放量和流式细胞术研究自噬在神经元转归中的作用。结果：Glu可显著增加神经元Che-1蛋白表达；过表达Che-1可减轻Glu所致神经元损伤，并减轻Glu所致神经元自噬；通过Rapamycin激活自噬可逆转Che-1对Glu所致神经元损伤的保护作用。结论：过表达Che-1蛋白可通过抑制神经元自噬对Glu所致神经元损伤发挥保护作用。 |
| 英文摘要: |
| ABSTRACT Objective: The aim of the present study was to observe the effect of glutamic acid (Glu) on the expression of Che-1 in cortical neurons, to investigate the effect of Che-1 overexpression on Glu-induced neuronal injury, and to elucidate the potential mechanism of Che-1-induced neuroprotection focusing on autophagy. Methods: After treatment with Glu, immunostaining and western blot were performed to detect the expression of Che-1 in neurons. Transfection with lentivirus was used to overexpress Che-1, and lactate dehydrogenase (LDH) release and flow cytometry were performed to measure neuronal injury. Immunostaining and western blot were used to determine autophagy in neurons. After treatment with the autophagy activator rapamycin, LDH release and flow cytometry were performed to investigate the protective effect of Che-1 against Glu. Results: Glu significantly increased the expression of Che-1 in cortical neurons. Overexpression of Che-1 alleviated the Glu-induced neuronal injury and reduced autophagy in neurons. The autophagy activator Che-1 partially prevented the protective effect of Che-1 against Glu in neurons. Conclusion: Overexpression of Che-1 could exert protective effect against Glu-induced neuronal injury through activating autophagy in cortical neurons. |
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