Study On The Mechanism Of Synaptic Plasticity Damage Of Hippocampal Neurons In Fetal Rats And The Intervention Of Traditional Chinese Medicine In A State Of Simulated Diabetes Complicated With Depression

Objective: To investigate the molecular mechanism of synaptic plasticity in hippocampal neurons leading to the occurrence of DD and the intervention of Chinese herbal compound ZGJTJYF,we study the hippocampal neurons in fetal rats under diabetes mellitus with depression.This study provides new ideas and targets for the research and prevention of the pathogenesis of DD.Methods: 1.The hippocampus of fetuses aged 7-10 days was isolated,purified and cultured,then neuron-specific enolase(NSE)immunocytochemical staining was performed for cell identification.2.The cultured hippocampal primary neurons were randomly divided into six groups: blank group,control group,model group,NR blocker group(MK-801),positive drug group(metformin + fluoxetine),ZGJTJYF drug group.According to the grouping situation,high glucose(15 mmol/L)and corticosterone(50 ?mol/L)was given to the model group;thecontrol group,the positive drug group and the ZGJTJYF drug group were separately add 10% volume blank serum,10% volume positive drug content serum,10% volume ZGJTJYF drug content serum to intervene after modeling,blank group was added 10% culture medium,and MK-801(10?mol/L)was added to blocker group,intervention for 18 h.The expression of CRHR1,GR,NR2 A,NR2B,CaMK II and SynGap proteins were detected by High Content Analysis(HCA).Whole cell patch clamp was used to record mEPSC in hippocampal neurons.The differences of mEPSC frequency and current amplitude in each group were compared.Results: 1.After being observed by inverted microscope and identified by NSE immunocytochemistry,hippocampal neuronal cells were confirmed,and the purity was over 95%.2.HCA analysis showed that the neurons of the blank group were intact in shape,full of cell bodies,and synapses were interwoven into neural networks.Compared with the blank group,hippocampal neurons in the control group and model group were atrophic,synaptic rupture or neural network disappeared,the fluorescence intensity of CRHR1,NR2 A,NR2B and CaMKII protein increased,and the fluorescence intensity of GR and SynGAP protein decreased(P<0.01).Compared with the control group,the synaptic connections of the NR blocker group/the positive drug group/the Zuogui Jiangtang Jieyufang group recovered,the fluorescence intensity of the NR2A/NR2B/CaMKII proteins decreased,and the fluorescence intensity of the SynGAP protein increased(P<0.01 or P<0.05);The fluorescence intensity of CRHR1 protein decreased in the positive drug group and the ZGJTJYF drug group,and the fluorescence intensity of GR protein increased(P<0.01),but no significant difference between the NR blocker group and the blank serum group.In patch clamp test,the frequency and current amplitude of mEPSC in the hippocampal neurons of the control group and the model group increased significantly compared with the blank group(P<0.01).Compared with the control group,the frequency and current amplitude of mEPSC in hippocampal neurons of the NR blocker group,positive drug group,and ZGJTJYF drug group significantly decreased(P<0.01).Conclusion: The Regulatory Effect of Zuo Gui Jiang Tang Jie Yu Fang on GR,CRHR1,NR2 A and NR2 B Proteins in Hippocampal Neurons under Simulated DD State,and the regulation of the synaptic plasticity and function of DD hippocampal neurons may be the protective mechanism against neuronal damage in DD rats.