SYK调控缺氧/缺糖损伤诱导的大脑皮质神经元凋亡及其机制

张志鑫; 崔应麟; 李彦杰; 任锟; 邢若星

doi:10.16016/j.1000-5404.201610168

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SYK调控缺氧/缺糖损伤诱导的大脑皮质神经元凋亡及其机制

基础医学 | 更新时间：2026-01-09

- SYK调控缺氧/缺糖损伤诱导的大脑皮质神经元凋亡及其机制
- Spleen tyrosine kinase regulates anoxia/hypoglycemia-induced apoptosis in cultured cerebral cortical neurons
- 第三军医大学学报 2017年39卷第13期页码：1381-1386
- 作者机构：
  
  450002 郑州，河南省中医院康复科
- 作者简介：
  
  [崔应麟, E-mail:junbosun@163.com CUI Yinglin, junbosun@163.com
- 基金信息：
- DOI：10.16016/j.1000-5404.201610168
  中图分类号： R345;R363.2;R743.302
- 收稿：2016-10-16，
  
  修回：2017-02-16，
  
  纸质出版：2017-07-15
- 稿件说明：
移动端阅览
张志鑫, 崔应麟, 李彦杰, 等. SYK调控缺氧/缺糖损伤诱导的大脑皮质神经元凋亡及其机制[J]. 第三军医大学学报, 2017,39(13):1381-1386.

Zhixin ZHANG, Yinglin CUI, Yanjie LI, et al. Spleen tyrosine kinase regulates anoxia/hypoglycemia-induced apoptosis in cultured cerebral cortical neurons[J]. Journal of Third Military Medical University, 2017, 39(13): 1381-1386.
张志鑫, 崔应麟, 李彦杰, 等. SYK调控缺氧/缺糖损伤诱导的大脑皮质神经元凋亡及其机制[J]. 第三军医大学学报, 2017,39(13):1381-1386. DOI： 10.16016/j.1000-5404.201610168.

Zhixin ZHANG, Yinglin CUI, Yanjie LI, et al. Spleen tyrosine kinase regulates anoxia/hypoglycemia-induced apoptosis in cultured cerebral cortical neurons[J]. Journal of Third Military Medical University, 2017, 39(13): 1381-1386. DOI： 10.16016/j.1000-5404.201610168.

摘要

目的

探讨抑制脾酪氨酸激酶(spleen tyrosine kinase，SYK)对缺氧/缺糖损伤诱导的大脑皮质神经元的凋亡的影响及相关机制。

方法

分离培养10只SD怀孕大鼠(190~230 g

胎龄16 d)胎鼠的大脑皮质神经元细胞并用MAP2免疫荧光法进行鉴定。建立缺氧/缺糖损伤的体外模型，caspase-3荧光检测试剂盒检测caspase-3活性，Annexin-Ⅴ FITC/PI法检测细胞凋亡，qRT-PCR检测SYK和凋亡相关的原癌基因Fra-1 mRNA的表达，Western Blot检测SYK、Bcl-2和Fra-1蛋白的表达。Control组：无转染处理的缺氧/缺糖损伤的神经元；SYK siRNA组：转染SYK siRNA的缺氧/缺糖损伤的神经元；SYK-Fra-1 siRNA组：同时转染SYK siRNA和Fra-1 siRNA的缺氧/缺糖损伤的神经元；non-specific siRNA组：转染non-specific siRNA的氧/缺糖损伤的神经元。

结果

MAP2免疫荧光鉴定为神经元细胞。缺氧/缺糖损伤诱导神经元SYK表达(

＜0.01)，RNA干扰抑制SYK表达后caspase-3活性(

＜0.05) 和细胞凋亡(

＜0.01) 均显著降低，但是Fra-1 mRNA (

＜0.01) 和蛋白(

＜0.05) 表达量明显上升。用RNA干扰技术同时抑制SYK和Fra-1后，caspase-3活性(

＜0.01) 显著上升但Bcl-2蛋白表达(

＜0.01) 显著降低。

结论

Fra-1介导了SYK对缺氧/缺糖损伤诱导的神经元凋亡的调节，为脑卒中所致的神经元缺血性损伤的治疗提供了新的靶点。

Abstract

Objective

To investigate the effect of inhibiting spleen tyrosine kinase (SYK) on apoptosis in cerebral cortical neuron induced by anoxia/hypoglycemia (A/H) injury and explore the related mechanism.

Methods

The fetal rat cerebral cortical neurons were isolated from 10 pregnant SD rats (weighting 190~230 g

gestational age of 16 d)

cultured and then identified by MAP2 immunofluorescence assay. The cerebral cortical neuron model with A/H injury was established

in vitro

. Then the cells were divided into A/H injured neurons (control group)

A/H injured neurons with SYK siRNA transfection (SYK siRNA group)

A/H injured neurons with co-transfection of SYK siRNA and Fra-1 siRNA (SYK-Fra-1 siRNA group)

and A/H injured neurons with non-specific siRNA transfection (non-specific siRNA group). Caspase-3 activity was detected by the caspase-3 kit. Cell apoptosis was determined by Annexin-Ⅴ FITC/PI assay. The mRNA expressions levels of SYK and apoptosis-related proto-oncogene Fra-1 were measured by qRT-PCR. The protein levels of SYK

Bcl-2 and Fra-1 were detected by using Western blotting.

Results

The neurons were identified by MAP2 immunofluorescence assay. Anoxia/hypoglycemia injury induced increased SYK expression in the neurons (

0.01). SYK silencing by RNA interference significantly reduced caspase-3 activity (

0.05) and cell apoptosis (

0.01)

while the mRNA (

0.01) and protein (

0.05) levels of Fra-1 were obviously up-regulated. Caspase-3 activity was significantly increased (

0.01) and Bcl-2 protein expression was markedly decreased (

0.01) in the cells after the co-silencing of SYK and Fra-1.

Conclusion

Fra-1 mediates SYK regulation in the apoptosis of cerebral cortical neurons induced by A/H injury

which provides the new target for A/H injury in neurons caused by cerebral stroke.

关键词

Keywords

references

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