越鞠丸合甘麦大枣汤加减对谷氨酸诱导的HT22细胞损伤模型的神经保护作用

葛渴敏; 王薇; 薛文达; 陈刚; 王福顺

doi:10.13422/j.cnki.syfjx.20190836

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越鞠丸合甘麦大枣汤加减对谷氨酸诱导的HT22细胞损伤模型的神经保护作用

经典名方 | 更新时间：2021-02-09

- 越鞠丸合甘麦大枣汤加减对谷氨酸诱导的HT22细胞损伤模型的神经保护作用
- Neuroprotective Effect of Modified Yuejuwan and Ganmai Dazao Tang on Glutamate-induced HT22 Cell Injury Model
- 中国实验方剂学杂志 2019年25卷第12期页码：22-27
- 作者机构：
  
  南京中医药大学　医学与生命科学学院，医学心理学院，基础医学院，南京　 210023
- 作者简介：
  
  葛渴敏，在读硕士，从事抑郁症中医研究，E-mail：1543803120@qq.com
  王福顺，博士，教授，从事神经生物学分子机制研究，E-mail：13814541138@163.com
- 基金信息：
  
  国家自然科学基金项目(81603497);江苏省自然科学基金项目(BK20151565)
- DOI：10.13422/j.cnki.syfjx.20190836
  中图分类号： R289; R322.8; R246.6; R749.4+ 1
- 收稿日期：2018-10-23，
  
  网络出版日期：2019-01-04，
  
  纸质出版日期：2019-06-20
- 稿件说明：
移动端阅览
葛渴敏, 王薇, 薛文达, 等. 越鞠丸合甘麦大枣汤加减对谷氨酸诱导的HT22细胞损伤模型的神经保护作用[J]. 中国实验方剂学杂志, 2019,25(12):22-27.

Ke-min GE, Wei WANG, Wen-da XUE, et al. Neuroprotective Effect of Modified Yuejuwan and Ganmai Dazao Tang on Glutamate-induced HT22 Cell Injury Model[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(12): 22-27.
葛渴敏, 王薇, 薛文达, 等. 越鞠丸合甘麦大枣汤加减对谷氨酸诱导的HT22细胞损伤模型的神经保护作用[J]. 中国实验方剂学杂志, 2019,25(12):22-27. DOI： 10.13422/j.cnki.syfjx.20190836.

Ke-min GE, Wei WANG, Wen-da XUE, et al. Neuroprotective Effect of Modified Yuejuwan and Ganmai Dazao Tang on Glutamate-induced HT22 Cell Injury Model[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(12): 22-27. DOI： 10.13422/j.cnki.syfjx.20190836.

摘要

目的：

探讨越鞠丸合甘麦大枣汤加减(YJGZ)对谷氨酸诱导的小鼠海马神经元细胞系(HT22)细胞损伤的神经保护作用。

方法：

HT22细胞系无菌培养，以高浓度谷氨酸建立细胞损伤模型，并制备YJGZ水提液和含药血清，分为正常组，模型组，YJGZ含药血清组(1%，5%，10%)，YJGZ水提液组(166 mg·L

－1

)，尼莫地平组(100 μmol·L

－1

)。噻唑蓝(MTT)比色法检测各组细胞存活率；酶联免疫吸附测定(ELISA)检测乳酸脱氢酶(LDH)释放率及细胞内活性氧自由基(ROS)的含量；蛋白免疫印迹法(Western blot)检测海马

-甲基-

-天冬氨酸受体亚单位2B蛋白(NR2B)，环磷腺苷效应元件结合蛋白(CREB)，磷酸化CREB(p-CREB)，细胞外调节蛋白激酶(ERK)，磷酸化ERK(p-ERK)的蛋白表达水平。

结果：

与正常组比较，模型组细胞存活率显著下降(

0.01)，乳酸脱氢酶(LDH)释放率明显增加(

0.05)，细胞内活性氧(ROS)水平明显增加(

0.05)，NR2B蛋白的表达明显升高(

0.05)，CREB，p-CREB，ERK，p-ERK蛋白表达水平明显降低(

0.05)；与模型组比较，YJGZ水提液组和尼莫地平组均能显著提高谷氨酸模型条件下HT22细胞的存活率(

0.01)，减少细胞损伤，减少LDH的释放率，降低细胞内ROS水平(

0.05，

0.01)，降低NR2B蛋白的表达水平(

0.05)，增加CREB，p-CREB，ERK，p-ERK蛋白的表达(

0.05)；YJGZ含药血清组与模型组比较，HT22细胞存活率并没有提高。

结论：

YJGZ水提液对谷氨酸诱导的HT22细胞损伤具有显著的保护作用。

Abstract

Objective:

To investigate the neuroprotective effect of modified Yuejuwan and Ganmai Dazao Tang (YJGZ) on glutamate-induced cell injury of mouse hippocampal neuron cell line (HT22).

Method:

The HT22 cell was cultured aseptically

and the cell injury model was established with high concentration of glutamate acid

YJGZ aqueous extract and drug-containing serum were prepared. Then experiment was divided into normal group

model group

YJGZ drug-containing serum groups (1%

10%)

YJGZ aqueous extract group (166 mg·L

-1

)

and nimodipine group (100 μmol·L

-1

). Methye thiazolye telrazlium (MTT) was used to detect the cell survival rate of each group

the level of lactate dehydrogenase (LDH) release and intracellular reactive oxygen species (ROS) were detected by enzyme-linked immunosorbent assay (ELISA). Western blot was used to detect the protein expression of hippocampal

-methyl-

-aspartate receptor subunit 2B protein (NR2B)

cyclic adenosine response element binding protein (CREB)

phosphorylated cyclic adenosine response element binding protein (p-CREB)

extracellular regulated protein kinase (ERK)

and phosphorylated extracellular regulated protein kinase (p-ERK).

Result:

As compared with normal group

the cell survival rate was significantly decreased in model group (

0.01)

LDH release rate was significantly increased (

0.05)

intracellular reactive oxygen species (ROS) level was increased (

0.05)

expression of NR2B protein was increased (

0.05)

and the protein expression levels of CREB

p-CREB

ERK and p-ERK were decreased (

0.05). As compared with model group

YJGZ aqueous extract group and nimodipine group can significantly improve the survival rate of HT22 cells under glutamate model conditions (

0.01)

reduce cell damage

reduce LDH release rate and intracellular ROS levels (

0.05

0.01)

decrease the expression level of NR2B protein (

0.05)

and increase the protein expression levels of CREB

p-CREB

ERK and p-ERK (

0.05). However

the survival rate of HT22 cells was not increased in the YJGZ-containing serum group as compared with model group.

Conclusion:

YJGZ aqueous extract has a significant protective effect on glutamate-induced HT22 cell injury.

关键词

Keywords

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