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酸枣仁汤对焦虑大鼠海马离子型谷氨酸受体表达及突触可塑性的影响

[Effect of Suanzaoren Decoction on expression of ionotropic glutamate receptors and synaptic plasticity in hippocampus of anxiety rats].

作者信息

Wang Hong-Kun, He Jin-Ming, Yan Yue-Heng, Wang Zi-Hao, Li Ruo-Xuan, Wang Yan-Yan

机构信息

Heilongjiang University of Chinese Medicine Harbin 150040, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2023 Oct;48(20):5583-5591. doi: 10.19540/j.cnki.cjcmm.20230704.401.

Abstract

This study investigated the effect of Suanzaoren Decoction on the expression of N-methyl-D-aspartate receptors(NMDAR) and α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptors(AMPAR) in the hippocampus and synaptic plasticity in rats with conditioned fear-induced anxiety. The effect of Suanzaoren Decoction on rat behaviors were evaluated through open field experiment, elevated plus maze experiment, and light/dark box experiment. Enzyme-linked immunosorbent assay(ELISA) was used to measure the levels of glutamate(Glu) and γ-aminobutyric acid(GABA) in the rat hippocampus. Real-time fluorescence quantitative PCR(qRT-PCR) and Western blot were employed to assess the gene and protein expression of ionotropic glutamate receptors in the hippocampal region. Transmission electron microscopy was utilized to observe the changes in the ultrastructure of synaptic neurons in the hippocampal region. Long-term potentiation(LTP) detection technique was employed to record the changes in population spike(PS) amplitude in the hippocampal region of mice in each group. The behavioral results showed that compared with the model group, the Suanzaoren Decoction group effectively increased the number of entries into open arms, time spent in open arms, percentage of time spent in open arms out of total movement time, number of entries into open arms out of total entries into both arms(P<0.01), and significantly increased the time spent in the light box and the number of shuttle crossings(P<0.01). There was an increasing trend in the number of grid crossings, entries into the center grid, and time spent in the center grid, indicating a significant anxiolytic effect. ELISA results showed that compared with the model group, the Suanzaoren Decoction group exhibited significantly reduced levels of Glu, Glu/GABA ratio(P<0.01), and significantly increased levels of GABA(P<0.01) in the rat hippocampus. Furthermore, Suanzaoren Decoction significantly decreased the gene and protein expression of NMDAR(GluN2B and GluN2A) and AMPAR(GluA1 and GluA2) compared with the model group. Transmission electron microscopy results demonstrated improvements in synapses, neuronal cells, and organelles in the hippocampal region of the Suanzaoren Decoction group compared with the model group. LTP detection results showed a significant increase in the PS amplitude changes in the hippocampal region of Suanzaoren Decoction group from 5 to 35 min compared with the model group(P<0.05, P<0.01). In conclusion, Suanzaoren Decoction exhibits significant anxiolytic effects, which may be attributed to the reduction in NMDAR and AMPAR expression levels and the improvement of synaptic plasticity.

摘要

本研究探讨了酸枣仁汤对条件性恐惧诱导焦虑大鼠海马中N-甲基-D-天冬氨酸受体(NMDAR)和α-氨基-3-羟基-5-甲基-4-异恶唑丙酸受体(AMPAR)表达及突触可塑性的影响。通过旷场实验、高架十字迷宫实验和明暗箱实验评估酸枣仁汤对大鼠行为的影响。采用酶联免疫吸附测定法(ELISA)检测大鼠海马中谷氨酸(Glu)和γ-氨基丁酸(GABA)的水平。运用实时荧光定量PCR(qRT-PCR)和蛋白质免疫印迹法评估海马区离子型谷氨酸受体的基因和蛋白表达。利用透射电子显微镜观察海马区突触神经元超微结构的变化。采用长时程增强(LTP)检测技术记录各组小鼠海马区群体峰电位(PS)幅度的变化。行为学结果显示,与模型组相比,酸枣仁汤组显著增加了进入开放臂的次数、在开放臂停留的时间、在开放臂停留时间占总运动时间的百分比、进入开放臂次数占进入双臂总次数的比例(P<0.01),并显著增加了在明箱停留的时间和穿梭次数(P<0.01)。方格穿越次数、进入中央方格次数和在中央方格停留时间呈增加趋势,表明具有显著的抗焦虑作用。ELISA结果显示,与模型组相比,酸枣仁汤组大鼠海马中Glu水平、Glu/GABA比值显著降低(P<0.01),GABA水平显著升高(P<0.01)。此外,与模型组相比,酸枣仁汤显著降低了NMDAR(GluN2B和GluNla)和AMPAR(GluA1和GluA2)的基因和蛋白表达。透射电子显微镜结果表明,与模型组相比,酸枣仁汤组海马区的突触、神经细胞和细胞器有所改善。LTP检测结果显示,与模型组相比,酸枣仁汤组海马区在5至35分钟时PS幅度变化显著增加(P<0.05,P<0.01)。综上所述,酸枣仁汤具有显著的抗焦虑作用,这可能归因于NMDAR和AMPAR表达水平的降低以及突触可塑性的改善。

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