Effect of Isopimpinellin on Expression of Inhibitory Neurotransmitter and Receptor Genes in Primary Hippocampal Neurons
10.13422/j.cnki.syfjx.20191302
- VernacularTitle: 异虎耳草素对原代海马神经元细胞抑制性神经递质及其受体基因表达的影响
- Author:
Yu-meng HAO
1
;
Ruo-yu WANG
1
;
Qi-ming ZHONG
1
;
Li-guo TONG
1
;
Mei-qing SONG
1
;
Qian YANG
1
;
Ma-li FENG
1
Author Information
1. Shanxi Academy of Traditional Chinese Medicine, Taiyuan 030012, China
- Publication Type:Research Article
- Keywords:
isopimpinellin;
hippocampal neuron;
inhibitory neurotransmitter;
cell apoptosis;
in vitro culture
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(14):93-98
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To observe the effect of isopimpinellin on primary hippocampal neuron cells γ-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 GABRA1,GABRA5,GABBR1, gamma-aminobutyric acid type B receptor genes (GABAB) GABRB2, 5-hydroxytryptamine 1A receptor (5-HT1A)5-HT1A(A),5-HT1A(B),5-HT1A(C) were detected by real-time quantitative PCR(Real-time PCR). Result:On the 7th 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 (P<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 (P<0.01). The mRNA expression levels of GABRA1,GABRA5,5-HT1A(A),5-HT1A(C) in the moderate-dose and high-dose isopimpinellin groups were significantly higher than those in the blank control group (P<0.05, P<0.01). The mRNA expression levels of GABBR1,5-HT1A(B) in the low-dose, moderate-dose and high-dose isopimpinellin groups were significantly higher than those in the blank control group (P<0.05, P<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.
