| Objective: To explore the possible affections of neuron survival rate and the change of BDNF and p-CREB induced by different concentration of Lamotrigine, Valproic acid and Phenobarbital.Methods: The hippocampus was harvested from newborn Sprague - Dawley rats. Primary hippocampal neuron cultures of 8 days in vitro were used and divided randomly into four groups: Valproic acid group, Phenobarbital group, lamotrigine group and 0.1%DMSO used as control group. Lamotrigine group was divided into low concentration (4 mg/ L) , middle concentration(8 mg/ L) ,high concentration(12 mg/ L). Valproic acid group was divided into low concentration (6.5mg/ L), middle concentration (13mg/ L), high concentration (26mg/ L). Phenobarbital group was divided into low concentration (6.5mg/ L), middle concentration (13mg/ L), high concentration (26mg/ L).The BDNF and p-CREB levels were determined by immunohistochemistry, and the cell survival rate was tested by MTT. Results: (1) 1 day after the drug administration, the light ring of neuron of the high concentration PB group was not bright and the neuritic network began to reduce. Others were the same as the control group. (2) 4 days after the drugs administration,in the high and the middle PB group, all concentration of VAP group and the LTG group, neurons demonstrated degenerative change, the neurites were thin the and the neuritic network began to reduce. The debrises of the neurons as well as bubbles in the neurons were seen in the high and middle concentration PB group and high concentration VAP group. (3) 7 days after the drugs administration, the loss of the fine neuritic network was seen, the neurites were swollen in the PB of high concentration group and the bubble of the neuron cell in the VAP of high concentration group was seen. The other neurons of the other drug groups were the same as the control group. (4)1 day after the drug administration, comparing with the control group, the survival of the neuron of high concentration of PB group decreased significantly(P<0.01). (5)After 4 days adding the drugs, comparing with the control group, the survival of the neuron of all the VAP group, LTG group, the middle and the high concentration PB group decreased significantly (P<0.05). (6) After 7 days adding the drugs, comparing with the control group, the survival of the neuron of all the VAP group, LTG group and PB group did not change significantly.(7)The cell survival rate decreased as the culture time going on with high concentration VAP group , high and middle concentration phenobarbital group and all concentration LTG group (P<0.05).(8)After 1 days adding the drugs, BDNF and p-CREB expressing on the cell were significantly different in the high concentration PB group(P<0.01).(9)After 4 days adding the drugs,comparing with the control group, BDNF and p-CREB expressing on the cell are significantly different in the middle and the high concentration PB group(P<0.05); comparing with the control group,low concentration VAP group and the middle VAP concentration group, the express of the BDNF in the high concentration of VAP, decreaed (P<0.05) and the express of the p-CREB decreaed(P<0.01); comparing with the control group and the middle LTG concentration group, the express of the BDNF in the high concentration of lamotrigine group,decreaed(P<0.01), comparing with the low LTG concentration group, the express of the BDNF in the high concentration of lamotrigine group,decreaed(P<0.05) and comparing with the control group,low concentration LTG group and the middle LTG concentration group, the express of the p-CREB in the high concentration of lamotrigine group,decreaed (P<0.05).(10)After 7 days adding the drugs, comparing with the control group, the expression of the BDNF and p-CREB in all the drug groups did not change significantly(P>0.05).(11)As the time goes by, all the groups, except the low PB concentration, the low and middle concentration VAP group ,the BDNF and p-CREB expressing on the cell are different significantly. Conclusion:(1)Except of PB, LTG and VAP can affect the cultured neurons'survival rate. (2)The damage caused by the AEDs has relationship with the drug concentration and the high concentration PB can affect the neuron survival rate soon after administration as well as the express of the BDNF and the p-CREB, significantly. (3) VAP and LTG can affect the neuron survival rate when keeping those drugs with the cultured neurons, but only high concentration can effect the BDNF and p-CREB express by changing the culture environment. (4) There is different between the clinical observing and the cell experiment but the mechanism should be discussed deeply further.
|
PDF Full Text Request