| Part 1 Effect of gossypol acetate on synaptic transmission of Schaffer col lateral-CA1 pathway in isolated hippocampal slicesObjectiveIn order to study the effect of gossypol acetate on the LTP of synaptic transmission in the Schaffer collateral-CA1 pathway on the isolated hippocampal slices of C57 mice,we conducted an in vitro LTP experiment,hoping to find out the specificity of gossypol acetate in the LTP of the Schaffer collateral-CA1 region effect.MethodsUsing the method of recording the field potential extracellularly,the mice were anesthetized with 10%chloral hydrate and the brain was quickly decapitated and placed on ice.After the mouse brain was properly trimmed with a blade,the brain slices were cut into 350μ M thick After incubation at room temperature for 30 minutes,incubate in a constant temperature water bath,and continue oxygenation.The stimulation electrode(concentric double-stranded electrode)is placed on the Schaffer collateral fibers emitted by the pyramidal cells in the hippocampal CA3 area and projected on the CA1 area,and the recording electrode(Glass electrode)placed in the pyramidal cell body layer of CA1 area[1].Stable fEPSP waveforms were induced with basic stimuli and recorded stably for 30 min,then high-frequency stimulation was given to induce the formation of LTP,and the effects of 5mg/kg,15mg/kg,and 30mg/kg gossypol acetate on LTP in mice were studied.ResultsCompared with the olive oil solvent control group,the gossypol acetate low,medium and high dose administration groups(5mg/kg,15mg/kg,30mg/kg)significantly increased the slope of fEPSP after high frequency stimulation,and the medium dose(15mg/kg)gossypol acetic acid administration group increased the slope of fEPSP after high frequency stimulation was higher than the low dose(5mg/kg)and high dose(15mg/kg)gossypol acetate groups.ConclusionGossypol acetate can promote LTP induced by high-frequency stimulation in the Schaffer lateral branch-CA1 area of mouse hippocampal slices;compared with the low-dose(5mg/kg)and high-dose(30mg/kg)gossypol acetate groups,The medium dose(15mg/kg)gossypol acetate has a stronger promoting effect on LTP,and the specific best dose of gossypol acetate needs further experimental verification.Part 2 Effect of gossypol acetate on Synaptic Transmission in the Anterior Peritoneal-Dentate Gyrus of the Hippocampal Entorhinal RegionObjectiveIn order to study the effect of gossypol acetate on the synaptic transmission of the pretransmembrane-dentate gyrus pathway in the rat olfactory region of the hippocampus,we conducted an in vivo LTP experiment,hoping to investigate the specific effect of gossypol acetate on the LTP of PP-DG.MethodsUsing in vivo field potential recording method,the rats were anesthetized with 25%urethane intraperitoneally,and their heads were fixed on the brain stereotaxic apparatus.According to the stereotaxic map of the rat brain,the PP and DG regions were located.The upper hole is used to place recording electrodes and bipolar stimulation electrodes[2].Stable PS waveforms were induced with basic stimuli and recorded stably for 30 min,after which high-frequency stimuli were induced to induce the formation of LTP[2],to study the low-dose(10mg/kg)and high-dose(30mg/kg)gossypol acetate The role of LTP in PP-DG area of rats.ResultsCompared with the blank control group,the low-dose(10mg/kg)gossypol acetate group significantly increased the PS wave amplitude after high-frequency stimulation.GonclusionLow-dose(10mg/kg)gossypol acetate has a significant effect on enhancing LTP in the brain circuit of rats.Part 3 Effect of gossypol acetate on the Expression of Synaptic Related Proteins and MeCP2ObjectiveIn order to study the effect of gossypol acetate on the expression of synaptic plasticity-related proteins in the rat brain,and to study the possible molecular mechanism of gossypol acetate improving synaptic transmission,we conducted proteomics and western blotting studies,hoping to explore gossypol acetate The specific role of LTP and its mechanism.MethodsThe rats were randomly divided into 5 groups,namely blank control group,olive oil control group,corn oil control group,olive oil gossypol acetate(30mg/kg)group and corn oil gossypol acetate(30mg/kg)group.The medicine was given by gavage every 48h for a total of 8 days.After 2 hours from the last dose,the brain was decapitated and the hippocampus and cortex were separated.Through proteomics experiments,the effect of gossypol acetate on the expression of synaptic plasticity-related proteins in the rat brain was studied.The mice were randomly divided into 3 groups:blank control group,solvent control group,and 15mg/kg gossypol acetate group,which were given by intragastric administration every 48h for a total of 8 days.The brain was decapitated 2 hours after the last administration Isolate mouse hippocampus and cortex.Through western blotting experiments,the effect of gossypol acetate on the expression of MeCP2 protein in mouse brain was studied.ResultsProteomics experiments showed that in the gossypol acetate administration group,there were changes in the expression of 20 synapse-related proteins,including KCNQ5.Many differentially expressed proteins were related to neuronal function and calcium ion regulation.Western blotting results showed that compared with the solvent control group,the expression of MeCP2 protein in the brain of the 15mg/kg gossypol acetate group increased significantly.Compared with the blank group,the expression of MeCP2 protein in the brain of the solvent control group did not change significantly.ConclusionThe results of proteomics experiments showed that the expression of 20 synaptic plasticity-related proteins in the gossypol acetate administration group changed.It may be that gossypol acetate regulates synaptic transmission and synaptic plasticity by regulating the changes of synapse-related proteins.The specific mechanism Further verification is required.Gossypol acetate(15mg/kg)can significantly increase the expression of MeCP2 in the mouse brain,and the solvent does not affect the expression of MeCP2 in the brain. |
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