Study On The Development Of Hippocampal Neurons In Fetal Rats With Gambogic Acid

Objective:The effects of gambogic acid on hippocampal neuron development in fetal rats were observed and evaluated.Cell proliferation was detected by cck-8,apoptosis was detected by flow cytometry,axon,dendritic length and number of branches were observed under microscope,and the expression level of RGS gene was detected,so as to investigate the protective effect of Gambogic acid(GA)on rat fetal rat neuron model.Methods:1.Establish the cells of the control group and the observation group: take the E16 day pregnant mice,isolate the sea horse tissue to culture the hippocampal neuron under the sterile condition,divide into the control group and the observation group,the control group does not carry on the treatment,the control group does not carry on the treatment,In observation group,normal hippocampal neurons were damaged by addition of A ?1-42..2.The cells were propagated to 96-well plate.The concentration gradient of GA,was 0,1,10,100,1 000 ? M respectively.72 hours later,the cells were digested and collected for CCK-8 assay,and the absorbance of each hole was measured under 450 nm.3.The final concentration of GA,in each group was 0?250 ? M by flow cytometry.48 hours later,the apoptosis of the cells in each group was observed.4.The axon,dendrite length and branching number of the four groups of cells(control group and A ? group cells without treatment and control group and A ? group cells added with GA)were observed under electron microscope.5.RT-q PCR,was used to observe the expression of RGS12 gene in the control cells and A ? cells treated with DMSO and GA dissolved in DMSO without any treatment.The expression level of RGS12 gene in the cells of each group was observed under the effect of GA.Results:1.The proliferation of hippocampal neurons in A ? group was significantly inhibited,while GA could effectively repair the damage induced by A ? in hippocampal neurons.2.With the increase of GA concentration,the apoptosis rate of the control group increased gradually,but there was no significant difference(P > 0.05).With the increase of GA concentration,the apoptosis rate of A ? group decreased gradually,and it was significantly different from that of 0 ?M GA group(P < 0.01).At the same concentration of GA,the apoptosis rate of the control group was significantly lower than that of the observation group.3.Compared with the control group,the number of dendrites and branches in A ? group decreased significantly,and the number of cell dendrites and branches in GA group decreased to a certain extent.Compared with A ? group,the number of dendrites and branches in A? GA group increased significantly.Compared with GA group,the number of dendritic cells in A ? GA group decreased significantly.4.After treated with GA,the relative expression level of m RNA of RGS12 in the A ?model cells was observed.Compared with the control group,the expression of RGS12 in the treated group was significantly up-regulated by RT-q PCR analysis,and the expression of RGS12 in the treated group was significantly higher than that in the control group(P < 0.05).In normal hippocampal neurons and hippocampal neuron A ?model cells,the expression of RGS12 in A ? model cells was significantly lower than that in normal hippocampal cells.Conclusion:Through this experiment,we studied that gambogic acid can promote cell proliferation,inhibit cell apoptosis,promote the growth of synapses,up-regulate the expression level of gene RGS,play the role of neurotrophic factors and protect the apoptosis mechanism of neurons,and provide further experimental research basis for clinical treatment.