数字中医药（英文）2020，Vol.3Issue(2)：80-95,16.DOI:10.1016/j.dcmed.2020.06.003
复方柴金解郁片通过改善海马神经元突触功能发挥抗抑郁作用
Compound Chai Jin Jie Yu Tablets, Acts as An Antidepressant by Promoting Synaptic Function in the Hippocampal Neurons
摘要
Abstract
Objective To investigate the effectiveness of compound Chai Jin Jie Yu Tablets (CJJYT) in ameliorating cognitive impairment associated with depression and its potential mechanism of action. Methods In vitro experiments, the hippocampus wasisolated from the whole brain of the fetal rat and cultured into hippocampal neuron cells. 50 μM corticosterone (CORT) was added to each group 18 h before the experiment for modeling depression, with the exception of the control group. After modeling, the blank serum group was added with 10％ blank serum, the CJJYT group and the venlafaxine group were added with the corresponding 10％ drug-containing serum, and the control group and the model group were added with equal volumes of culture medium. The intracellular Ca2 + mean fluorescence intensity, miniature excitatory postsynaptic current (mEPSC) current amplitude, and frequency of different hippocampal neurons were evaluated as indicators of synaptic function in the hippocampal neurons. In addition, the expression of synaptic plasticity related proteins, synaptophysin-α (SYN-α), N-methyl-D-aspartate receptor 2A (NR2A), N-methyl-D-aspartate receptor 2B (NR2B), post synaptic density 95 protein (PSD-95), calcium/calmodulin dependent protein kinaseⅡ(CaMKⅡ) and synaptic associated protein (SynGAP) were detected in the hippocampal neurons by immunofluorescence staining and high content analysis (HCA) system. Then, reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA expression levels of SYN-α, NR2A, NR2B, PSD-95, CaMKⅡ and SynGAP. For in vivo experiments, except for those in the blank control group, all rats were treated within a single cage for chronic unpredictable stress-induced depression modeling and subjected to corresponding drug interventions. Behavioral tests were used to detect depressive behavior and determine learning, memory and other cognitive abilities, whereas enzyme-linked immunosorbent assay (ELISA) was used to detect the CORT levels. Golgi-Cox staining was used to observe changes in the synaptic morphology of parahippocampal gyrus CA1 area (CA1) and dentategyrus (DG).Results In vitro, CJJYT treatment reduced the intracellular Ca2 +mean flurorescence intensityin the hippocampal neurons (P < 0.05), causing a reduction in the frequency and current amplitude of mEPSC(P <0.05), and thus inhibited the excessive activation of post-synaptic receptors. CJJYT treatment reduced the protein and mRNA expression of SYN-α, NR2A, NR2B and PSD-95 in the hippocampal neurons(P <0.05), increased the mRNA and protein expression of CaMKⅡ and SynGAP(P <0.05), and thereby improved the synaptic plasticity of the hippocampal neurons. In vivo, CJJYT intervention improved sucrose preference, voluntary activity, learning and memory ability of Morris water maze test, and suppressed appetite (P <0.05), and increased the despair feeling of forced swimming test(P <0.05). The CORT level wasreduced(P <0.05), leading to the repair of synaptic damage in the hippocampal neurons. Conclusions CJJYT can improve the synaptic function of hippocampal neurons and has obvious protective effects on neurons. It can repair the structural damage in the hippocampal neurons, improving the cognitive ability of the depressed model rats. The mechanism ofCJJYT improving cognition in depressed rats may be related to the transmission and function of SYN-α/NR and its downstream neurotransmitters.关键词
复方柴金解郁片/抑郁症/SYN-α/NR/突触可塑性/认知Key words
Compound Chai Jin Jie Yu Tablets (CJJYT)/Depression/SYN-α/NR/Synaptic plasticity/Cognition引用本文复制引用
李姿蓉,韩远山,吴梦瑶,刘检,金狮,张熙,王宇红..复方柴金解郁片通过改善海马神经元突触功能发挥抗抑郁作用[J].数字中医药（英文）,2020,3(2):80-95,16.基金项目
We thank for the funding support from the National Major New Drug Development Project (No. 2017ZX09309026) and Provincial Department of Graduate Research Innovation Project of Hunan (No. CX20190565). （No. 2017ZX09309026）
