MicroRNA-124 promotes neurite elongation and motor function recovery in mice with cerebral ischemia injury by targeting PTEN

Jingda ZHANG; Shuhui DAI

doi:10.16016/j.2097-0927.202202025

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MicroRNA-124 promotes neurite elongation and motor function recovery in mice with cerebral ischemia injury by targeting PTEN

Neurosciences | 更新时间：2026-01-09

- MicroRNA-124 promotes neurite elongation and motor function recovery in mice with cerebral ischemia injury by targeting PTEN
- Journal of Army Medical University Vol. 44, Issue 14, Pages: 1478-1485(2022)
- 作者机构：
  
  1.571199 海南，海南医学院公共卫生学院
  2.710032 西安，空军军医大学西京医院神经外科
- 作者简介：
- 基金信息：
  
  the National Natural Science Foundation for Young Scholars of China(81901186)
- DOI：10.16016/j.2097-0927.202202025
  CLC： R363.22;R394.1;R743.31
- Received：10 February 2021，
  
  Revised：2021-03-05，
  
  Published：30 July 2022
- 稿件说明：
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Jingda ZHANG, Shuhui DAI. MicroRNA-124 promotes neurite elongation and motor function recovery in mice with cerebral ischemia injury by targeting PTEN[J]. Journal of Army Medical University, 2022, 44(14): 1478-1485.
DOI：

Jingda ZHANG, Shuhui DAI. MicroRNA-124 promotes neurite elongation and motor function recovery in mice with cerebral ischemia injury by targeting PTEN[J]. Journal of Army Medical University, 2022, 44(14): 1478-1485. DOI： 10.16016/j.2097-0927.202202025.

摘要

目的

探究MicroRNA-124(miR-124)对脑缺血损伤小鼠神经功能的改善作用及其内在机制。

方法

① 取32只C57小鼠随机分为假手术组和脑缺血组(

=16)，TTC染色和神经行为学评分检测小鼠脑损伤后3天脑梗死面积和运动情况，RT-qPCR检测miR-124在两组间的表达变化。②培养原代神经元细胞，通过氧糖剥夺模拟缺血再灌注损伤，观察氧糖剥夺后0、72 h miR-124的变化趋势。③另取32只C57小鼠随机分为脑缺血组和脑缺血+miR-124组(

=16)，TTC染色、转棒实验和肢体放置实验检测小鼠的运动功能改善情况。④细胞实验分为氧糖剥夺组和氧糖剥夺+miR-124组，通过免疫荧光实验检测两组神经元轴突的生长情况。Targetscan数据库预测miR-124下游靶基因，双荧光素酶基因报告和Western blot在原代神经元等上验证对照组和miR-124组下游靶基因第10号染色体缺失的磷酸酶及张力蛋白同源基因(the phosphate and tensin homolog deleted on chromosome ten

PTEN)的表达变化。上调PTEN，检测氧糖剥夺+miR-124和氧糖剥夺+miR-124+PTEN两组神经元轴突的生长情况。⑤在体实验中检测脑缺血+miR-124组和脑缺血+miR-124+Pten组(

=8)转棒实验和肢体放置情况。

结果

脑缺血再灌注后，小鼠脑梗死面积约达到40%(

0.001)，小鼠的神经认知功能显著降低(

0.001)，已达到重度认知功能损害；miR-124的表达较对照组明显升高(

0.05)；而进一步过表达miR-124能够促进氧糖剥夺后神经元轴突的生长(

0.05)，显著缩小脑梗死面积(

0.05)，且促进动物运动能力的恢复和改善(

0.05)；Targetscan数据库预测miR-124下游靶基因为PTEN，双荧光素酶基因报告和Western blot都证实miR-124能有效下调PTEN的表达(

0.05)；最后，上调PTEN逆转了miR-124促进神经元轴突的生长以及改善小鼠运动能力的效应(

0.05)。

结论

miR-124通过下调PTEN促进脑缺血小鼠神经元轴突生长，且有助于小鼠运动功能的恢复。

Abstract

Objective

To explore the effect of microRNA-124 (miR-124) on the improvement of neurological function in mice with cerebral ischemia injury and its underlying mechanism.

Methods

① A total of 32 C57 mice were randomly divided into sham group and cerebral ischemia group (

=16). The cerebral infarction area was detected by TTC staining and motor status of the mice was evaluated with neurobehavioral scoring in 3 d after brain injury. The expression change of miR-124 was detected by RT-qPCR and compare

d between the 2 groups. ② Primary neurons were isolated from fetal mice and cultured

and ischemia-reperfusion injury was simulated by oxygen glucose deprivation. The changes of miR-124 at 0 and 72 h after oxygen glucose deprivation were observed. ③ Another 32 C57 mice were randomly divided into cerebral ischemia group and cerebral ischemia+miR-124 group (

=16). TTC staining

rotating rod test and limb placement test were performed respectively to detect whether the motor function of the mice was improved. ④ The primary neurons were divided into oxygen-glucose deprivation group and oxygen-glucose deprivation+miR-124 group. The neurite elongation in the 2 groups was detected by immunofluorescence assay. The downstream target gene of miR-124 was predicted with Targetscan database. Double luciferase gene reporter assay and Western blotting were used to verify the effectiveness of miR-124 regulation on phosphatase and tensin homolog deleted on chromosome 10 (PTEN) in the control and miR-124 groups. Then

the effect of PTEN up-regulation on neurite elongation was observed in the oxygen glucose deprivation+miR-124 cells and oxygen glucose deprivation+miR-124+PTEN groups. ⑤ Rotating rod test and limb placement test were performed again for motor functions in the mice of cerebral ischemia+miR-124 group and cerebral ischemia+miR-124+PTEN group (

=8).

Results

Cerebral ischemia-reperfusion resulted in the area of cerebral infarction about 40% (

0.001) and significantly reduced neurocognitive function in mice (

0.001)

reaching the condition of severe cognitive impairment. The expression of miR-124 was significantly higher in the cerebral ischemia group than the control group (

0.05). Overexpression of miR-124 could promote the growth of neuronal axons after oxygen glucose deprivation (

0.05)

reduce the area of cerebral infarction (

0.05)

and thu

s promote the recovery and improvement of animal motor ability (

0.05). With the aid of Targetscan database

PTEN was predicted as the downstream target of miR-124

and double luciferase gene reporter assay and Western blotting confirmed that miR-124 could effectively down-regulate the expression of PTEN (

0.05); Finally

up-regulation of PTEN reversed the effect of miR-124 on promoting the growth of neuronal axons and improving the motor ability of mice (

0.05).

Conclusion

MiR-124 promotes neuronal axonal growth and contributes to the recovery of motor function in mice with cerebral ischemia by down-regulating PTEN.

关键词

Keywords

references

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