中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2014, Vol. 9 ›› Issue (17): 1599-1605.doi: 10.4103/1673-5374.141811

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

ATP诱导神经细胞损伤1 h内可发生自噬：对抗凋亡的神经保护效应

  

  • 收稿日期:2014-08-17 出版日期:2014-09-16 发布日期:2014-09-16

Autophagy occurs within an hour of adenosine triphosphate treatment after nerve cell damage: the neuroprotective effects of adenosine triphosphate against apoptosis

Na Lu 1, Yong Wang 2, Baoying Wang 1, Honggang Zhao 1, Dongliang Li 1   

  1. 1 Department of Physiology and Neurobiology, Xinxiang Medical University, Xinxiang, Henan Province, China
    2 Department of Human Anatomy, Xinxiang Medical University, Xinxiang, Henan Province, China
    3 Department of Laboratory Animal Center, Xinxiang Medical University, Xinxiang, Henan Province, China
  • Received:2014-08-17 Online:2014-09-16 Published:2014-09-16
  • Contact: Dongliang Li, Department of Physiology and Neurobiology, Xinxiang Medical University, Xinxiang 453003, Henan Province, China, xyldl8@126.com.

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摘要：

当中枢神经系统出现缺氧缺血、炎症损伤后，受损细胞释放大量ATP，可引起神经元继发性死亡。自噬(autophagy)作为细胞死亡形式之一，具有脑保护作用。实验通过MTT法、MDC 染色法、流式细胞仪、蛋白质印迹、RT-PCR法检测不同浓度(2，4，6，8，10 mmol/L)和不同作用时间(1，2，3，6 h)的ATP(6 mmol/L)对与神经元功能相类似的SH-SY5Y神经细胞凋亡及自噬的影响。结果表明，胞外高浓度ATP能够诱导SH-SY5Y神经细胞自噬和凋亡，最先出现自噬增强现象，ATP干预1 h自噬增强达高峰；细胞凋亡高潮在后，3 h达高峰，且持续到6 h，随着ATP作用时间的延长，凋亡占主导地位。说明SH-SY5Y神经细胞在ATP干预的1 h内，很可能优先启动细胞自噬来对抗凋亡，以此发挥保护神经细胞损伤的作用

关键词: 神经再生, 神经元, ATP, SH-SY5Y神经细胞, 自噬, 凋亡, 细胞培养, 酰戊二胺, 流式细胞术, 细胞活力, Bcl-2, Bax, Beclin 1, 神经元损伤, 国家自然科学基金

Abstract:

After hypoxia, ischemia, or inflammatory injuries to the central nervous system, the damaged cells release a large amount of adenosine triphosphate, which may cause secondary neuronal death. Autophagy is a form of cell death that also has neuroprotective effects. Cell Counting Kit assay, monodansylcadaverine staining, flow cytometry, western blotting, and real-time PCR were used to determine the effects of exogenous adenosine triphosphate treatment at different concentrations (2, 4, 6, 8, 10 mmol/L) over time (1, 2, 3, and 6 hours) on the apoptosis and autophagy of SH-SY5Y cells. High concentrations of extracellular adenosine triphosphate induced autophagy and apoptosis of SH-SY5Y cells. The enhanced autophagy first appeared, and peaked at 1 hour after treatment with adenosine triphosphate. Cell apoptosis peaked at 3 hours, and persisted through 6 hours. With prolonged exposure to the adenosine triphosphate treatment, the fraction of apoptotic cells increased. These data suggest that the SH-SY5Y neural cells initiated autophagy against apoptosis within an hour of adenosine triphosphate treatment to protect themselves against injury.

Key words: nerve regeneration, neurons, adenosine triphosphate, SH-SY5Y cells, autophagy, apoptosis, cell culture, monodansylcadaverine, flow cytometry, cell viability, Bcl-2, Bax, Beclin 1, neuronal damage, NSFC grant, neural regeneration