Effects Of Phenobarbital On Apoptosis Of Neurons And Neurogliocyte In Frontal Lobe Parietal Lobe And Cerebellum Of Rats

Objective: Epilepsy is a group of conditions involving transient disturbance in cerebral function caused by abnormal neuronal discharges .It is a common disease in neurology. Drug therapy has remained the main approach so far for treating patients with epilepsy. Although phenobarbital(PB) is not the most optimal drug, it has the advantages of definite effect cheap and abundant. Recently doctors find PB has lots of function beyond antiepileptic effect.In department of pediatrics, it is used to treat neonatal hypoxie-ischemic encephalopathy, prevent preterm infant intracranial hemorrhage and the relapse of febrile convulsions. In gastroenterology department, it is used to treat acute icteric hepatitis and biliary ascariasis. In department of neurology, it can also be used to treat severe epidemic encephalitis B, subarachnoid hemorrhage, vertigo, neurodystrophic edema, and prevent post craniotomy seizure[1]. Thus phenobarbital is still used in many epilepsy patients although multiple drugs available.So far, systemic researches have been finished on brain injury of infant rat induced by antiepileptic drugs administration. Antiepileptic drugs lead to apoptosis of neuron through affecting the expression of gene, ion channels, neurotransmitters and the second messenger systems. Antiepileptic drugs also infect infant rat brain by influencing the migration of cell, differentiation, synapse formation and synaptic plasticity, nerve fiber myelinogenesis. Because the same targets regulate brain processes essential both for propagation of seizures and for learning, memory and emotional behavior, the normal function was suppressed when controlling seizure. Reasearches about effects of phenobarbital on apoptosis of neuron and neurogliocyte in frontal lobe, parietal lobe and cerebellum of rats are rare. It is worth exploring whether the apoptosis of neuron and neurogliocyte in these areas associate with the adverse effects in nervous system.This passage aims at phenobarbital's adverse effects in nervous system. Phenobarbital was given to healthy rats. Neuron and neurogliocyte apoptosis in frontal lobe, parietal lobe and cerebellum of rats was detected by terminal deoxynucleotidyl transfers-mediated biotinylated deoxyuridine triphosphate nickel end labeling. Expression of apoptosis-related proteins Bcl-2 and Bax were detected by immunohistochemistry stain.Methods: 30 healthy Sprague Dawley rats aged ten weeks were divided into five groups. Group A was treated with normal saline (2ml/100g/d) by stomach lavaging, it was divided to two subgroups, and half of rats were killed separately at 15 day and 30 day. Group B was treated with PB (30mg/kg/d) for fifteen days. Group C was treated with PB (30mg/kg/d)for thirty days. Group D was treated with PB (75mg/kg/d) for fifteen days. Group E was treated with PB (75mg/kg/d) for thirty days. Then rats were sacrificed and brains were made to paraffin blocks. Use the TUNEL and immunohistochemistry stain to detect the level of apoptosis of neuron and neurogliocyte and expression of proteins Bcl-2 and Bax in frontal lobe, parietal lobe and cerebellum of rats under optical microscope.Statics: Use One-Way ANOVA to compare the difference between groups, the comparison between two groups was achieved by SNK method. P=0.05 as the check standard.Results:1 Group E has lots of TUNEL positive cells in frontal lobe. A small quantity of TUNEL positive cells were observed in other groups. P<0.01through group E comparing with group D, group C, group A long-term subgroup in frontal lobe, it has statistically significant. P>0.05 through group E comparing with group D, group C, group A long-term subgroup in parietal lobe and cerebellum, it is not statistically significant. P>0.05 through group D comparing with group A short-term subgroup, group B in frontal lobe, parietal lobe and cerebellum, it is not statistically significant. P>0.05 through group B comparing with group C, group A short-term subgroup, and group C comparing with group A long-term subgroup in frontal lobe, parietal lobe and cerebellum, it is not statistically significant. P>0.05 through group A short-term subgroup comparing with long-term subgroup in frontal lobe, parietal lobe and cerebellum, it is not statistically significant.2 The expression of Bax and Bcl-2 exist in both cytolymph and nucleus and presenting sepia color.Group A has small quantity of positive expression of Bax and Bcl-2 in two subgroups. P>0.05 through group A short-term subgroup comparing with long-term subgroup in frontal lobe, parietal lobe and cerebellum, it is not statistically significant.There are small quantity of positive expression of Bax and Bcl-2 in Group B and Group C. P>0.05 through group B comparing with group C in frontal lobe, parietal lobe and cerebellum, it is not statistically significant. P>0.05 through group B comparing with group A short-term subgroup and group C comparing with group A long-term subgroup in frontal lobe, parietal lobe and cerebellum, it is not statistically significant.Group E has a lot of positive expression of Bax in frontal lobe. P<0.01through group E comparing with group D, group C, group A long-term subgroup in frontal lobe, it has statistically significant. There are small quantity of positive expression of Bcl-2 in Group E and Group D in frontal lobe, P>0.05through Group E omparing with Group D, group C, group A long-term subgroup in frontal lobe, it has statistically significant. There are small quantity of positive expression of Bcl-2 in Group E and Group D in parietal lobe and cerebellum. P>0.05through Group E omparing with Group D, group C, group A long-term subgroup in parietal lobe and cerebellum, it has statistically significant. P>0.05through positive expression of Bax and Bcl-2 in Group D omparing with group A short-term subgroup, group B in frontal lobe, parietal lobe and cerebellum, it is not statistically significant. Conclusions:1 Treatment concentration PB(30mg/kg/d) have no significance effect to the apoptosis of neuron and gliocyte in frontal parietal and cerebellum.2 Administration high concentration PB(75mg/kg/d) for a longer time make the expression of Bax and the apoptosis of neuron and gliocyte in frontal increase.It suggests PB may promote apoptosis of neuron and gliocyte by increasing the expression of Bax.3 It has no significant effect to the apoptosis of neuron and gliocyte in parietal and cerebellum after long-term and high concentration PB administration.