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 (19): 1770-1778.doi: 10.4103/1673-5374.143421

• 原著:退行性病与再生 • 上一篇    

激活的腺苷酸活化蛋白激酶促进了肌萎缩侧索硬化模型小鼠胚胎神经干细胞凋亡

  

  • 收稿日期:2014-07-22 出版日期:2014-10-15 发布日期:2014-10-15
  • 基金资助:

    国家自然科学基金（81030019）

Adenosine monophosphate-activated protein kinase activation enhances embryonic neural stem cell apoptosis in a mouse model of amyotrophic lateral sclerosis

Yanling Sui, Zichun Zhao, Rong Liu, Bin Cai, Dongsheng Fan   

  1. Department of Neurology, Peking University Third Hospital, Beijing, China
  • Received:2014-07-22 Online:2014-10-15 Published:2014-10-15
  • Contact: Dongsheng Fan, M.D., Ph.D., Department of Neurology, Peking University Third Hospital, 49 North Garden Road, Haidian District, Beijing 100191, China, wshns99@163.com.
  • Supported by:

    This work was supported by a grant from the National Natural Sciences Foundation of China, No. 81030019.

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摘要：

研究胚胎期神经干细胞的异常变化对揭示肌萎缩侧索硬化发病机制非常重要，我们假设SOD1G93A个体的胚胎神经干细胞遇氧化应激更易受损，可能会增加后期发生神经退行性疾病的易感性。将过氧化氢作用于SOD1G93A小鼠和野生型SOD1WT小鼠胚胎神经干细胞，分别采用CCK-8细胞计数法和流式细胞术检测了神经干细胞的活力及凋亡。然后采用蛋白免疫印迹法检测了腺苷酸活化蛋白激酶α、Pax3和p53的活性或表达。SOD1G93A小鼠胚胎神经干细胞遇过氧化氢时较SOD1WT小鼠胚胎神经干细胞更容易发生凋亡。SOD1G93A小鼠胚胎神经干中腺苷酸活化蛋白激酶α磷酸化水平较SOD1WT小鼠中磷酸化水平高，Pax3表达量与腺苷酸活化蛋白激酶α磷酸化水平呈负相关，而p53表达量与腺苷酸活化蛋白激酶α磷酸化水平变化趋势相同。除此之外，腺苷酸活化蛋白激酶抑制剂compound C减弱了过氧化氢对神经干细胞的促凋亡作用。结果说明，SOD1G93A小鼠胚胎神经干细胞在氧化应激条件下更易凋亡，该作用主要由腺苷酸活化蛋白激酶通路中的Pax3和p53蛋白介导。

关键词: 神经再生, 神经退行性变, 胚胎神经干细胞, AMPKα, 配对盒3, p53, SOD1G93A小鼠, 肌萎缩侧索硬化, 氧化应激, 过氧化氢, 神经细胞凋亡, 国家自然科学基金

Abstract:

Alterations in embryonic neural stem cells play crucial roles in the pathogenesis of amyotrophic lateral sclerosis. We hypothesized that embryonic neural stem cells from SOD1G93A individuals might be more susceptible to oxidative injury, resulting in a propensity for neurodegeneration at later stages. In this study, embryonic neural stem cells obtained from human superoxide dismutase 1 mutant (SOD1G93A) and wild-type (SOD1WT) mouse models were exposed to H2O2. We assayed cell viability with mitochondrial succinic dehydrogenase colorimetric reagent, and measured cell apoptosis by flow cytometry. Moreover, we evaluated the expression of the adenosine monophosphate-activated protein kinase (AMPK) α-subunit, paired box 3 (Pax3) protein, and p53 in western blot analyses. Compared with SOD1WT cells, SOD1G93A embryonic neural stem cells were more likely to undergo H2O2-induced apoptosis. Phosphorylation of AMPKα in SOD1G93A cells was higher than that in SOD1WT cells. Pax3 expression was inversely correlated with the phosphorylation levels of AMPKα. p53 protein levels were also correlated with AMPKα phosphorylation levels. Compound C, an inhibitor of AMPKα, attenuated the effects of H2O2. These results suggest that embryonic neural stem cells from SOD1G93A mice are more susceptible to apoptosis in the presence of oxidative stress compared with those from wild-type controls, and the effects are mainly mediated by Pax3 and p53 in the AMPKα pathway.

Key words: nerve regeneration, neuroderegeneration, embryonic neural stem cells, adenosine monophosphate-activated protein kinase α, paired box 3, p53, SOD1G93A mouse, amyotrophic lateral sclerosis, oxidative stress, hydrogen peroxide, apoptosis, NSFC grants, neural regeneration