摘要
探讨酸枣仁汤对条件性恐惧致焦虑大鼠海马离子型谷氨酸受体N-甲基-D-天冬氨酸受体(N-methyl-D-aspartate receptor,NMDAR)和α-氨基-3-羟基-5-甲基-4-异唑丙酸受体(α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor,AMPAR)的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考察酸枣仁汤对焦虑大鼠行为学的影响;利用酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测大鼠海马中谷氨酸(glutamate,Glu)和γ-氨基丁酸(γ-aminobutyric acid,GABA)含量;采用实时荧光定量PCR(real-time fluorescence quantitative PCR,qRT-PCR)、蛋白免疫印迹法(Western blot)检测大鼠海马区离子型谷氨酸受体的基因及蛋白表达;采用透射电镜技术观察大鼠海马区神经突触超微结构的变化,并通过在体长时程增强(long-term potentiation,LTP)检测技术记录各组大鼠海马区的群峰电位(polulation spike,PS)幅值变化率。行为学结果显示,与模型组相比,酸枣仁汤能有效增加焦虑大鼠的开臂次数、开臂运动时间、开臂占总运动时间百分比、开臂占入臂总次数百分比(P<0.01),显著升高明箱时间和穿梭次数(P<0.01),跨格次数、进入中央格次数、进入中央格时间有增加趋势,表现出明显的抗焦虑作用;ELISA结果显示,与模型组相比,酸枣仁汤组大鼠海马组织中的Glu含量和Glu/GABA比值显著降低(P<0.01),GABA含量显著升高(P<0.01);此外,与模型组相比,酸枣仁汤能明显降低NMDAR(GluN2B、GluN2A)、AMPAR(GluA1、GluA2)的基因和蛋白表达。透射电镜结果表明,酸枣仁汤组海马区的突触、神经元细胞及细胞器较模型组均有改善。LTP检测结果显示,与模型组比较,酸枣仁汤组大鼠海马区的PS幅值变化率在5~35 min显著增加(P<0.05,P<0.01)。综上,酸枣仁汤具有显著的抗焦虑作用,其作用机制可能与减少NMDAR、AMPAR表达水平以及改善神经突触可塑性有关。
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
作者
王红坤
何金铭
颜岳衡
汪子皓
李若暄
王艳艳
WANG Hong-kun;HE Jin-ming;YAN Yue-heng;WANG Zi-hao;LI Ruo-xuan;WANG Yan-yan(Heilongjiang University of Chinese Medicine,Harbin 150040,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2023年第20期5583-5591,共9页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(82003974)
黑龙江省自然科学基金项目(LH2022H086)
哈尔滨市科技局项目(2013RFQXJ070)
黑龙江省中医药科研项目(LH2023)。
