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载脂蛋白E对机械损伤后神经元细胞内钙水平及细胞凋亡的影响。

Effects of ApoE on intracellular calcium levels and apoptosis of neurons after mechanical injury.

作者信息

Jiang L, Zhong J, Dou X, Cheng C, Huang Z, Sun X

机构信息

Department of Neurosurgery, The First Affiliated Hospital of Chongqing Medical University, PR China.

Chongqing Medical University, PR China.

出版信息

Neuroscience. 2015 Aug 20;301:375-83. doi: 10.1016/j.neuroscience.2015.06.005. Epub 2015 Jun 11.

Abstract

OBJECTIVE

The current study aimed to explore the effects of apolipoprotein e (ApoE) on intracellular calcium ([Ca(2+)]i) and apoptosis of neurons after mechanical injury in vitro.

METHODS

A neuron mechanical injury model was established after primary neurons obtained from APOE knockout and wild-type (WT) mice, and four experimental groups were generated: Group-ApoE4, Group-ApoE3, Group-ApoE(-) and Group-WT. Recombinant ApoE4 and ApoE3 were added to Group-ApoE4 and Group-ApoE3 respectively, and Group-ApoE(-) and Group-WT were control groups. Intracellular calcium was labeled by fluo-3/AM and examined using laser scanning confocal microscope and flow cytometry, and the apoptosis of neurons was also evaluated.

RESULTS

The intracellular calcium levels and apoptosis rates of mice neurons were significantly higher in Group-ApoE4 than in Group-ApoE3 and Group-WT after mechanical injury. However, without mechanical injury on neurons, no significant differences in intracellular calcium levels and apoptosis rates were found among all four experimental groups. The effects of ApoE4 on intracellular calcium levels and apoptosis rates of injured neurons were partly decreased by EGTA treatment.

CONCLUSION

Compared with ApoE3-treatment and WT neurons, ApoE4 caused higher intracellular calcium levels and apoptosis rates of neurons after mechanical injury. This suggested APOE polymorphisms may affect neuron apoptosis after mechanical injury through different influences on intracellular calcium levels.

摘要

目的

本研究旨在探讨载脂蛋白E(ApoE)对体外机械损伤后神经元细胞内钙([Ca(2+)]i)及细胞凋亡的影响。

方法

从载脂蛋白E基因敲除小鼠和野生型(WT)小鼠获取原代神经元后建立神经元机械损伤模型,分为四组:ApoE4组、ApoE3组、ApoE(-)组和WT组。分别向ApoE4组和ApoE3组添加重组ApoE4和ApoE3,ApoE(-)组和WT组为对照组。用Fluo-3/AM标记细胞内钙,采用激光扫描共聚焦显微镜和流式细胞术检测,并评估神经元的凋亡情况。

结果

机械损伤后,ApoE4组小鼠神经元的细胞内钙水平和凋亡率显著高于ApoE3组和WT组。然而,在未对神经元进行机械损伤时,四个实验组的细胞内钙水平和凋亡率均无显著差异。EGTA处理可部分降低ApoE4对损伤神经元细胞内钙水平和凋亡率的影响。

结论

与ApoE3处理组和WT神经元相比,ApoE4在机械损伤后导致神经元细胞内钙水平和凋亡率更高。这表明载脂蛋白E基因多态性可能通过对细胞内钙水平的不同影响,从而影响机械损伤后神经元的凋亡。

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