Study Of SERCA Expression In Intractable Epilepsy Cell Model And The Effect Of Gastrodin Intervention

Objective: To study the expression of SERCA2(ATP2a2)gene and protein in refractory epilepsy cell model,and the effect of gastrodin intervention on the expression of SERCA2(ATP2a2)in refractory epilepsy cell model,thus further exploring the antiepileptic mechanism of gastrodin.Methods: Newborn rats,after sterilizated with 75% alcohol,quickly be decapitated the brain and isolated the bilateral hippocampus.Culturing the hippocampal neurons with the neuron-specific serum-free culture base(neurobasal Plus media)and B-27 supplements in vitro.To identify neurons by immunofluorescence of anti-NeuN and then calculate the purity on the 5th day.7th Day,the cultured neurons were randomly grouped into the normal control group(C),model group(M),model + 1 ?M gastrodin group(G1),model + 10 ?M gastrodin group(G2),model + 100 ?M gastrodin group(G3),Model + 1 mM gastrodin group(G4).Observing the morphological changes of neurons in each group through inverted phase contrast microscope.The expression of ATP2a2(SERCA2)mRNA was detected by real-time fluorescence quantitative PCR(RT-PCR).And the expression changes of ATP2a2 protein and c-PARP protein were detected by western blot technique.Results:(1)The neuronal fusion and death were found in the refractory epilepsy cell model.(2)In the model group,the expression of apoptotic marker c-PARP protein was significantly increased(P<0.01),and gastrodin could decrease the expression of apoptotic c-PARP(P<0.05).(3)Compared with the control group,the expression of ATP2a2(SERCA2)mRNA in the model group decreased(p<0.05),and that of the 100 uM gastrodin group was significantly increased(p<0.05).(4)ATP2a2 is mainly expressed in endoplasmic reticulum.Average light density of normal control group(1.0039±0.0027),model group(0.4371359 ± 0.1522186),model+100 ?M gastrodin group(0.9770±0.0323).There was a statistically significant difference between the normal control group and the model group(p<0.05).And the difference between the model group and the model+100 ?M gastrodin group was statistically significant(p<0.05).Conclusion:1.SERCA2(ATP2a2)gene and protein expression reduction and neural network changes occur in refractory epilepsy models.2.There is neuronal lesion and apoptosis in the cell model of refractory epilepsy.3.Gastrodin can raise the expression of SERCA2(ATP2a2)gene and protein in refractory epilepsy cell model.4.Gastrodin can ease the apoptosis of refractory epilepsy cell model.5.Gastrodin can alleviate the apoptosis of refractory epilepsy cell model through up-regulating SERCA2(ATP2a2)expression.