Targeting PTEN to regulate autophagy and promote the repair of injured neurons
Targeting PTEN to regulate autophagy and promote the repair of injured neurons
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DOI：
10.1016/j.brainresbull.2020.10.008
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发表时间：
2020-12-01
影响因子：
3.8
通讯作者：
Shen, Jianhong
中科院分区：
文献类型：
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作者：
Yu, Haoyuan;Shao, Junjie;Shen, Jianhong
自噬对神经元损伤的影响可能是积极的，也可能是有害的。通过建立胎鼠皮层神经元液压休克损伤模型，在损伤后不同时间点用双过氧（picolinoto）钒酸二钾[bpv（pic）]抑制PTEN表达，观察神经元损伤后自噬水平的变化。根据bpv（pic）抑制自噬的时间点将神经元分为干预组，正常神经元和损伤神经元设为对照组。通过免疫荧光染色评估各组神经元的生长。Western blot分析自噬相关蛋白LC3-II和LC3-I的表达。RT-PCR检测PTEN、mTOR和Beclin-1的表达。电镜观察正常组、损伤对照组、24 h、36 h干预组自噬体数量，发现神经元损伤后自噬增强，bpv（pic）干预后LC 3-II水平显著降低。伤后24 h和30 h，伤后对照组的生长发育不如正常组，但经bpv（pic）干预后，生长发育得到改善。Western blot分析显示，损伤后LC3-II和LC3-II/LC3-I比值增加，电镜下可见明显的自噬诱导。这些效果通过RT-PCR分析得到证实。综上所述，这些数据表明，神经元损伤后自噬被激活，而bpv（pic）可抑制自噬，从而促进损伤神经元的修复。
The effects of autophagy on neuronal damage can be positive or detrimental negative. Through establishing a model of fetal rat cortical neuron hydraulic shock injury, dipotassium bisperoxo (picolinoto) oxovanadate (V) [bpv(pic)] was used to inhibit PTEN at different time points post-injury and autophagy level after neuronal injury was assessed. Neurons were divided into several intervention groups according to the time point at which bpv (pic) was used to inhibit autophagy, normal neurons and injuried neurons were set as two control groups. Growth of neurons in each group was assessed through immunofluorescence staining. Expression of the autophagy-related proteins LC3-II and LC3-I was analyzed by western blot. Expression of PTEN, mTOR and Beclin-1 was detected by RT-PCR. The number of autophagosomes in the normal group, injury control group and 24 h, 36 h intervention groups were assessed by electron microscope.We found that autophagy was enhanced after neuronal injury and that the levels of LC3-II was significantly reduced by bpv (pic) intervention. The growth of the injury control groups was worse than normal groups, while improved through bpv(pic) intervention at 24 h and 30 h after injured. Western blot analysis showed that the LC3-II and LC3-II/LC3-I ratios of cells increased post-injury, and autophagy induction was evident by electron microscopy. These effects were confirmed by RT-PCR analysis. Taken together, these data suggest that autophagy is activated after injury in neurons while can be inhibited by bpv(pic) administration and then promote the repair of injured neurons.