Effects Of Ganoderic Acid A On The Expression Of Calcium-sensitive Receptors And Apoptotic Proteins In Hippocampal Neurons Incubated With Magnesiu-free Extracellular Fluid

Objective: Detect the morphological changes,survival rate,apoptosis rate and Ca SR and Bax、Bcl-2、Caspase-3 protein expression of hippocampal neurons induced by ganoderma lucidum A in magnesium-free extracellular fluid,to explore the effect of Ganoderma lucidum acid A on incubating hippocampal neurons without magnesium extracellular fluid.Methods: Take the hippocampal tissue of 1-3-day-old SD rats and primary culture of hippocampal neurons in vitro.The hippocampal neurons were cultured until the 12 th day and random Ly divided into seven groups according to the random number table,including the control group,hippocampal neurons in magnesium-free extracellular fluid-induced group,hippocampal neurons in sodium valproate group,hippocampal neurons in ganoderma acid A group,hippocampal neurons in the magnesium-free extracellular fluid in the ganoderma acid A and spermine group,hippocampal neurons in the magnesium-free extracellular fluid in the ganoderma acid A and NPS2390 group,hippocampal neurons in the magnesium-free extracellular fluid in the ganoderma acid A and NPS2390 and spermine group.The growth status of hippocampal neurons and morphological changes of each group was observed by inverted microscope;The survival rate and apoptosis rate of neurons in each group were detected by MTT colorimetry,TUNEL staining method.Western blot protein expression of Ca SR and Bax、Bcl-2、Caspase-3 in each group was detected by Western blot.Results: Observed from the inverted microscope,hippocampal neurons cultured to 12 days,neuronal processes crisscrossed,and neurons formed extensive synaptic connections between cells,typical neural nets;Hippocampal neurons in magnesium-free extracellular fluidinduced group were crumpled,rounded,synapses became less and some synaptic connections were broken,while hippocampal neurons in ganoderma acid A group had more synaptic connections.MTT colorimetry showed that the survival rate of hippocampal neurons in magnesium-free extracellular fluid-induced group was significantly lower than that in normal group(P<0.01);The survival rate of hippocampal neurons in sodium valproate group and ganoderma acid A group decreased significantly compared with that in magnesium-free extracellular fluid-induced group(P<0.01).TUNEL staining showed that,the apoptotic cells were brownish,cell shrinkage and apoptotic bodies were observed.The apoptosis rate of hippocampal neurons in magnesium-free extracellular fluid-induced group was significantly higher than that in normal group(P<0.01);The apoptosis rate of hippocampal neurons in sodium valproate group and ganoderma acid A group decreased significantly compared with that in magnesium-free extracellular fluid-induced group(P<0.01).Western blot showed that there was no significant difference between the two groups(P>0.05);The expression of Ca SR、Bax、Caspase-3 protein in the group induced by magnesium-free extracellular fluid was higher than that in the normal group(P>0.05),the expression of Bcl-2 was decreased(P<0.05);Compared with the magnesium-free extracellular fluid-induced group,the Ca SR、Caspase-3 protein expression in the ganoderma acid A group decreased that was no statistical difference(P>0.05),there was no significant difference in Bax protein expression(P>0.05),the expression of Bcl-2protein increased that was no statistical difference(P>0.05).Conclsions: The magnesium-free extracellular fluid can induce the injury of hippocampal neurons;ganoderma acid A can decreased the expression of Ca SR after magnesium-free extracellular fluid induction;ganoderma acid A can improve the survival rate of hippocampal neurons and reduce apoptosis rate after magnesium-free extracellular fluid induction,and has antiepileptic effect.