白藜芦醇经PI3K/Akt途径促进脑出血后大鼠轴突再生

王冠宇; 刘畅; 周昌龙; 夏小辉; 张光伟; 谭兴卫; 崔敏; 贺学农

doi:10.16016/j.1000-5404.201904006

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白藜芦醇经PI3K/Akt途径促进脑出血后大鼠轴突再生

基础医学 | 更新时间：2026-01-09

- 白藜芦醇经PI3K/Akt途径促进脑出血后大鼠轴突再生
- Resveratrol promotes axonal regeneration via PI3K/Akt pathway in rats after intracerebral hemorrhage
- 第三军医大学学报 2019年41卷第16期页码：1527-1537
- 作者机构：
  
  402160 重庆，重庆医科大学附属永川医院神经外科
- 作者简介：
  
  [ 贺学农，E-mail:cqychxn1965@yahoo.cn HE Xuenong, E-mail: cqychxn1965@yahoo.cn
- 基金信息：
  
  重庆市科学技术委员会项目(CSTC2018jscx-msybX0125)
- DOI：10.16016/j.1000-5404.201904006
  中图分类号： R282.71; R285.5; R743.34
- 收稿：2019-04-01，
  
  修回：2019-05-04，
  
  纸质出版：2019-08-30
- 稿件说明：
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王冠宇, 刘畅, 周昌龙, 等. 白藜芦醇经PI3K/Akt途径促进脑出血后大鼠轴突再生[J]. 第三军医大学学报, 2019,41(16):1527-1537.

Guanyu WANG, Chang LIU, Changlong ZHOU, et al. Resveratrol promotes axonal regeneration via PI3K/Akt pathway in rats after intracerebral hemorrhage[J]. Journal of Third Military Medical University, 2019, 41(16): 1527-1537.
王冠宇, 刘畅, 周昌龙, 等. 白藜芦醇经PI3K/Akt途径促进脑出血后大鼠轴突再生[J]. 第三军医大学学报, 2019,41(16):1527-1537. DOI： 10.16016/j.1000-5404.201904006.

Guanyu WANG, Chang LIU, Changlong ZHOU, et al. Resveratrol promotes axonal regeneration via PI3K/Akt pathway in rats after intracerebral hemorrhage[J]. Journal of Third Military Medical University, 2019, 41(16): 1527-1537. DOI： 10.16016/j.1000-5404.201904006.

摘要

目的

探讨白藜芦醇通过PI3K/Akt途径调控AMPKα通路，促进脑出血后大鼠轴突再生的机制。

方法

选取220~250 g雄性SD大鼠，按随机数字表法分为假手术组、脑出血组、小剂量白藜芦醇组(20 mg/kg)、大剂量白藜芦醇组(40 mg/kg)、抑制剂组(PI3K/Akt抑制剂LY294002)，采用自体血注射法对SD大鼠建立脑出血模型。评估大鼠神经功能缺损，HE染色观察脑组织病理变化，ELISA检测各组大鼠氧化应激相关指标，Western blot、免疫组化和免疫荧光检测各组大鼠神经丝蛋白200(neurofilament 200，NF200)、微管相关蛋白-2(microtubule-associated protein-2，MAP-2)、髓鞘碱性蛋白(myelin basic protein，MBP)、生长相关蛋白(growth-associated protein 43，GAP43)及蛋白激酶B(protein kinase B，PKB or Akt)、腺苷酸活化蛋白激酶α(AMP-activated protein kinase alpha

AMPKα)、糖原合酶激酶-3β(glycogen synthase kinase 3 beta，GSK-3β)的表达。

结果

脑出血后大鼠的神经功能缺损程度显著升高，HE染色可见明显病理损伤，氧化应激产物水平明显增高，使用白藜芦醇干预之后损伤程度减轻，且大剂量白藜芦醇组效果明显优于小剂量白藜芦醇组(

0.05)。Western blot与免疫组化检测显示：与脑出血组比较，注射白藜芦醇后大鼠脑出血区域组织中NF200和MAP-2表达明显升高，且大剂量白藜芦醇组效果明显优于小剂量白藜芦醇组(

0.05)。Western blot与免疫荧光结果显示：注射白藜芦醇后MBP表达显著高于脑出血组，大剂量白藜芦醇组效果明显优于小剂量白藜芦醇组(

0.05)，且白藜芦醇通过AMPK作用于轴突。对通路蛋白进一步进行Western blot检测，与脑出血组比较，注射白藜芦醇后大鼠脑出血区域组织中p-AMPKα、p-GSK-3β表达降低，NF200、MAP-2、MBP、GAP43和p-Akt表达升高，且大剂量白藜芦醇组效果明显优于小剂量白藜芦醇组(

0.05)，使用抑制剂后可逆转白藜芦醇干预后的效应。

结论

白藜芦醇可促进脑出血后大鼠的轴突再生，对神经功能的恢复有明显改善，其机制可能与通过PI3K/Akt途径抑制AMPK通路有关。

Abstract

Objective

To investigate the mechanism by which resveratrol (RES) promotes axonal regeneration in rats after intracerebral hemorrhage (ICH) and explore the role of the PI3K/Akt pathway that regulates the AMPKα pathway.

Methods

A total of 126 male adult SD rats were randomized into sham- operated group (

=30)

ICH group (

=30)

low-dose (20 mg/kg) RES group (

=30)

high-dose (40 mg/kg) RES group and PI3K/Akt inhibitor (LY294002) group (

=6). Rat models of ICH were established by stereotaxic injection of autologous blood into the basal ganglia with subsequent trea

tments as indicated. The neurological deficits of the rats were evaluated

and the pathological changes in the brain tissues were observed with HE staining. The parameters of oxidative stress in each group were detected using ELISA. Western blotting

immunohistochemistry and immunofluorescence assay were performed to detect the expressions of neurofilament 200 (NF200)

microtubule-associated protein-2 (MAP-2)

myelin basic protein (MBP)

growth-associated protein 43 (GAP43)

protein kinase B (PKB or Akt)

AMP-activated protein kinase alpha (AMPKα) and glycogen synthase kinase 3 beta (GSK-3β) in the brain tissue.

Results

The rats exhibited significant neurological deficits

obvious brain pathologies

and elevated oxidative stress level following ICH. Intervention with RES

especially at the high dose

obviously alleviated the injuries of the brain tissue. Western blotting and immunohistochemistry showed that compared with those in ICH group

the expression of NF200 and MAP-2 increased significantly in the hemorrhagic region in rats receiving RES treatment

and the effect was more obvious in the high-dose group than in the low-dose group (

0.05). Immunofluorescence assay showed that RES treatment of the rats

especially at the high dose

obviously enhanced MBP expression in the brain tissue as compared with that in ICH group (

0.05)

and AMPK was found to mediate the effect of RES on the axons. The results of Western blotting further confirmed that RES treatment significantly decreased the expressions of p-AMPKα and p-GSK-3β and increased the expressions of NF200

MAP-2

MBP

GAP43 and p-Akt in the hemorrhagic brain tissues of the rats

as compared with those in ICH group

and high-dose RES produced significantly stronger effects on these proteins (

0.05). These effects of RES were obviously reversed by the application of PI3K/Akt inhibitor in the rat models of ICH.

Conclusion

RES can promote axonal regeneration and neurological function recovery in rats after ICH possibly by down-regulating the AMPK pathway through the PI3K/Akt pathway.

关键词

Keywords

references

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