Modified Shuyuwan Ameliorate Neuronal Apoptosis of APP/PS1 Mice Via Akt/GSK3β/Nrf2 Pathway

Jing QIU; Zi-hu TAN; Qiong YANG; Xiao-yan WANG; Fang HUANG

doi:10.13422/j.cnki.syfjx.20192140

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Modified Shuyuwan Ameliorate Neuronal Apoptosis of APP/PS1 Mice Via Akt/GSK3β/Nrf2 Pathway

Pharmacology | 更新时间：2021-02-09

- Modified Shuyuwan Ameliorate Neuronal Apoptosis of APP/PS1 Mice Via Akt/GSK3β/Nrf2 Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 21, Pages: 38-44(2019)
- 作者机构：
  
  1.湖北中医药大学，武汉　 430065；
  2.湖北省中医院，武汉　 430061
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20192140
  CLC： R2-0; R285; R289; R338
- Published：05 November 2019，
  
  Published Online：19 July 2019，
  
  Received：18 June 2019，
- 稿件说明：
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Jing QIU, Zi-hu TAN, Qiong YANG, et al. Modified Shuyuwan Ameliorate Neuronal Apoptosis of APP/PS1 Mice Via Akt/GSK3β/Nrf2 Pathway. [J]. Chinese Journal of Experimental Traditional Medical Formulae 25(21):38-44(2019)
DOI：

Jing QIU, Zi-hu TAN, Qiong YANG, et al. Modified Shuyuwan Ameliorate Neuronal Apoptosis of APP/PS1 Mice Via Akt/GSK3β/Nrf2 Pathway. [J]. Chinese Journal of Experimental Traditional Medical Formulae 25(21):38-44(2019) DOI： 10.13422/j.cnki.syfjx.20192140.

摘要

目的：

探讨加减薯蓣丸对APP/PS1模型小鼠神经保护的效应和作用机制。

方法：

5月龄雄性APP/PS1小鼠20只和野生型小鼠10只，分为空白组、模型组、加减薯蓣丸组(14 g·kg

－1

·d

－1

)，灌胃28 d，空白组、模型组给予等体积生理盐水；APP/PS1背景和野生型背景原代神经元，分为空白组、模型组、加减薯蓣丸组、衣霉素组和磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(Akt)抑制剂组，空白组、模型组给予10%空白血清，加减薯蓣丸组给予5%加减薯蓣丸含药血清干预，衣霉素组和PI3K/Akt抑制剂组分别在加减薯蓣丸基础上加用2 mg·L

－1

的衣霉素和10 μmol·L

－1

的LY294002。采用Morris水迷宫检测小鼠学习记忆能力；采用蛋白免疫印迹法(Western blot)检测海马核因子E2相关因子2(Nrf2)蛋白表达；Western blot检测内质网应激相关蛋白糖调节蛋白78(GRP78)，蛋白激酶样内质网激酶(PERK)，磷酸化(p-)PERK，凋亡通路蛋白真核生物的翻译起始因子2的

亚基(eIF2

)，p-eIF2

，增强子结合蛋白同源蛋白(CHOP)，半胱氨酸蛋白酶-3(Caspase-3)和p-Akt，Akt，糖原激酶3

(GSK3

)，Nrf2蛋白表达。

结果：

体内实验中，与空白组比较，模型组APP/PS1小鼠学习记忆能力显著下降(

0.01)，海马Nrf2表达显著下调(

0.01)；与模型组比较，加减薯蓣丸干预4周后，加减薯蓣丸组APP/PS1小鼠学习记忆能力明显提升，海马Nrf2表达水平显著增加(

0.01)。体外实验中，与空白组比较，模型组GRP78，p-PERK/PERK，p-eIF2

/eIF2

，CHOP，cleaved Caspase-3蛋白表达显著增加(

0.01)；与模型组比较，加减薯蓣丸含药血清明显降低GRP78，p-PERK/PERK，p-eIF2

/eIF2

，CHOP，cleaved Caspase-3蛋白表达(

0.05，

0.01)；而衣霉素抑制加减薯蓣丸对内质网应激诱导凋亡的保护效应(

0.05)；与空白组比较，模型组p-Akt/Akt，Nrf2蛋白表达显著下降，GSK-3

表达显著增加(

0.01)；与模型组比较，加减薯蓣丸含药血清干预后p-Akt/Akt，Nrf2蛋白表达显著增加，GSK-3

表达明显降低(

0.05，

0.01)；LY294002抑制加减薯蓣丸对p-Akt/Akt，Nrf2和GSK3

蛋白表达的影响(

0.05，

0.01)。

结论：

加减薯蓣丸通过PI3K/Akt/GSK3

信号通路上调Nrf2蛋白表达，减轻内质网应激诱导的神经元凋亡，改善APP/PS1模型小鼠学习记忆能力。

Abstract

Objective:

To explore the effect and mechanism of modified Shuyuwan neuroprotection in APP/PS1 model mice .

Method:

Selecting 20 male APP/PS1 mice of 5 months old and 10 wild type mice.The mice were divided into blank group

model group and modified Shuyuwan group(14 g·kg

-1

·d

-1

)

drug delivery for 28 days

and blank group and model group were given the same amount of normal saline

APP/PS1 background primary neuron model and wild type primary neurons were divided into blank group

model group and modified Shuyuwan group

tunicamycin group

phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) inhibitor group.The blank group and model group were given 10%blank serum

the modified Shuyuwan group was given 5%modified Shuyuwan-containing serum

the tunicamycin group and the PI3K/Akt inhibitor group were respectively added with 2 mg·L

-1

tunicamycin and 10 μmol·L

-1

LY294002 on the basis of 5%modified Shuyuwan-containing serum.The spatial learning and memory ability of mice was measured by Morris water maze

and Western blot was used to detect nuclear factor erythroid-2 related factor 2(Nrf2) protein expression in hippocampus.Western blot was used to detect the protein expression of endoplasmic reticulum stress related proteins glucose regulatory protein 78(GRP78)

protein kinase-like endoplasmic reticulum kinase(PERK)

phosphorylation (p-)PERK and apoptosis expression of the pathway proteins eukaryotic translation initiation factor 2

(eIF2

)

p-eIF2

enhancer binding protein homologous protein(CHOP)

and cysteinyl aspartate apecific proteinase 3 (Caspase-3)

p-Akt

Akt

Glycogen Synthase kinase-3

(GSK3

)

Nrf2.

Result:

In vivo

experiment

compared with blank group

the learning and memory ability of APP/PS1 mice in the model group was impaired(

0.01)

and the expression level of Nrf2 in the hippocampus was decreased(

0.01). Compared with model group

after 4 weeks of modified Shuyuwan intervention

the learning and memory ability of APP/PS1 mice in the modified Shuyuwan group was improved

and the expression level of Nrf2 in the hippocampus was significantly increased(

0.01).

In vitro

experiment

Western blot analysis showed that compared with the blank group

the expression of GRP78

p-PERK/PERK

p-eIF2

/eIF2

CHOP

and cleaved Caspase-3 proteins was increased in the model group(

0.01). Compared with model group

modified Shuyuwan-containing serum intervention significantly reduced the expression of GRP78

p-PERK/PERK

p-eIF2

/eIF2

CHOP

and cleaved Caspase-3 proteins(

0.05

0.01). Whereas tunicamycin inhibited the protection effect of SHhuyuwan on the endoplasmic reticulum and ERS-induced apoptosis(

0.05). Western blot results showed that compared with blank group

the expression of p-Akt/Akt and Nrf2 protein was significantly decreased and the expression of GSK-3

was increased in the model group(

0.01). Compared with model group

the expression of p-Akt/Akt and Nrf2 protein was significantly increased and the expression of GSK-3

was decreased after modified Shuyuwan-containing serum intervention(

0.05

0.01). The effect of modified Shuyuwan on the expression of p-Akt

Nrf2 and GSK3

protein was inhibited by LY294002.(

0.05

0.01).

Conclusion:

Modified Shuyuwan can increase Nrf2 protein expression through PI3K/Akt/GSK3

signaling pathway

reduce neuronal apoptosis induced by endoplasmic reticulum stress

improve the learning and memory ability of APP/PS1 model mice.

关键词

阿尔茨海默病加减薯蓣丸核因子E2相关因子2内质网应激凋亡

Keywords

Alzheimer's diseasemodified Shuyuwannuclear factor erythroid-2 related factor 2endoplasmic reticulum stressapoptosis

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