Effect of Isopimpinellin on Expression of Inhibitory Neurotransmitter and Receptor Genes in Primary Hippocampal Neurons

Yu-meng HAO; Ruo-yu WANG; Qi-ming ZHONG; Li-guo TONG; Mei-qing SONG; Qian YANG; Ma-li FENG

doi:10.13422/j.cnki.syfjx.20191302

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Effect of Isopimpinellin on Expression of Inhibitory Neurotransmitter and Receptor Genes in Primary Hippocampal Neurons

Pharmacology | 更新时间：2021-02-09

- Effect of Isopimpinellin on Expression of Inhibitory Neurotransmitter and Receptor Genes in Primary Hippocampal Neurons
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 14, Pages: 93-98(2019)
- 作者机构：
  
  山西省中医药研究院，太原　 030012
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20191302
  CLC： R2-0; R22; R285.5
- Received：10 December 2018，
  
  Published Online：19 March 2019，
  
  Published：20 July 2019
- 稿件说明：
移动端阅览
Yu-meng HAO, Ruo-yu WANG, Qi-ming ZHONG, et al. Effect of Isopimpinellin on Expression of Inhibitory Neurotransmitter and Receptor Genes in Primary Hippocampal Neurons[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(14): 93-98.
DOI：

Yu-meng HAO, Ruo-yu WANG, Qi-ming ZHONG, et al. Effect of Isopimpinellin on Expression of Inhibitory Neurotransmitter and Receptor Genes in Primary Hippocampal Neurons[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(14): 93-98. DOI： 10.13422/j.cnki.syfjx.20191302.

摘要

目的：

观察异虎耳草素对原代海马神经元细胞

-氨基丁酸(GABA)，5-羟色胺(5-HT)含量及受体基因表达的影响，探讨其催眠作用机制。

方法：

体外培养SD乳鼠原代海马神经元细胞，在细胞生长状态最佳时，经神经元特异性烯醇化酶(NSE)免疫组化鉴定纯度并给药。分为空白组，地西泮组(25 mg·L

－1

)，异虎耳草素低、中、高剂量组(5，10，20 mg·L

－1

)，共5组，给药24 h后流式细胞术检测海马神经元细胞早期凋亡率，酶联免疫吸附测定(ELISA)检测细胞上清液GABA，5-HT含量，实时荧光定量聚合酶链式反应(Real-time PCR)检测细胞内

-氨基丁酸A型受体(GABAA)基因GABRA

，GABRA

，GABRB

，

-氨基丁酸 B型受体(GABAB)基因GABBR1，5-羟色胺1A型受体(5-HT1

)基因5-HT1

A(A)

，5-HT1

A(B)

，5-HT1

A(C)

表达的变化。

结果：

第7天海马神经元细胞生长状态良好，鉴定纯度

90%；与空白组比较，海马神经元细胞早期凋亡率异虎耳草素中、高剂量组显著降低(

0.01)，GABA含量异虎耳草素高剂量组显著升高(

0.01)，GABRA

，GABRA

，5-HT1

A(A)

，5-HT1

A(C)

mRNA表达异虎耳草素中、高剂量组显著升高(

0.05，

0.01)，GABBR

，5-HT1

A(B)

mRNA表达异虎耳草素各组均显著升高(

0.05，

0.01)。

结论：

异虎耳草素催眠作用机制可能与抑制海马神经元细胞凋亡，升高抑制性神经递质GABA含量，上调GABA与5-HT相关受体基因表达有关。

Abstract

Objective:

To observe the effect of isopimpinellin on primary hippocampal neuron cells

-aminobutyric acid (GABA)

5-hydroxytryptamine (5-HT) and receptor genes expressions

in order to explore its hypnotic mechanism.

Method:

The primary hippocampal neurons of neonatal Sprague-Dawley rats were cultured

in vitro

. And subsequent experiments were conducted in the optimal state of cell growth

and the purity was identified by immunohistochemistry of neuron-specific enolase. Hippocampal neurons were randomly divided into five groups

namely blank control group

diazepam group (25 mg·L

-1

)

and low-dose (5 mg·L

-1

)

moderate-dose (10 mg·L

-1

) and high-dose (20 mg·L

-1

) isopimpinellin groups. Early apoptosis of hippocampus neuron cells were detected using flow cytometry technique after 24 h administration

and the changes in the levels of GABA and 5-HT were detected using enzyme-linked immunosorbent. The changes in mRNA expressions of receptor genes relating to gamma-aminobutyric acid type A receptor(GABAA) genes GABRA

GABRA

GABBR

gamma-aminobutyric acid type B receptor genes (GABAB) GABRB

5-hydroxytryptamine 1A receptor (5-HT1

)5-HT1

A(A)

5-HT1

A(B)

5-HT1

A(C)

were detected by real-time quantitative PCR(Real-time PCR).

Result:

On the 7

day

the hippocampal neurons grew in a good condition

and the purity was above 90%. Apoptosis rates of hippocampal neurons in the low-dose and moderate-dose groups were significantly lower than that in the blank control group (

0.01). The level of GABA secreted by hippocampal neurons in the high-dose isopimpinellin group were significantly higher than that in the blank control group (

0.01). The mRNA expression levels of GABRA

GABRA

5-HT1

A(A)

5-HT1

A(C)

in the moderate-dose and high-dose isopimpinellin groups were significantly higher than those in the blank control group (

0.05

0.01). The mRNA expression levels of GABBR

5-HT1

A(B)

in the low-dose

moderate-dose and high-dose isopimpinellin groups were significantly higher than those in the blank control group (

0.05

0.01).

Conclusion:

The hypnotic mechanism of isopimpinellin may be related to the inhibition of hippocampal neuron apoptosis

the increase of the content of inhibitory neurotransmitter GABA

and the up-regulation of GABA and 5-HT-related receptor genes.

关键词

Keywords

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