| ObjectiveTo explore the anti epileptic effect and underlying mechanism of the Shi Gan powder on general conditions、behaviouristics、hippocampal neurons ion channel current average and Brain Derived Neurotropic Factor(BDNF)in pentetrazole induced epileptic rat model.Providing necessary theoretical foundation for treating and preventing epilepsy.Methods110grown-upSD rats were selected,15 rats were randomly assigned to control group,the rest were lighted with PTZ.Recording the behavioural changes after molding.Western medicine group were given sodium valproate15.75mg/Kg and Chinese tranditional medicine group received Shi Gan powder0.4g/Kg、1.8g/Kg、1.6g/Kg via intragastric administration,respectively.Control group and model group were administrated the distilled water 3ml/kg.After 21 days treatment,observing the behavioral change、weights change、hippocampal pathologicaltissue change by Nissl’s staining、BDNF content change in hippocampal by Immunohistochemical method and ion channel currents changes in voltage dependent potassium and sodium channels in epileptic rat model.Results1.The behavioral changes were performed before and after the treatment of rats:①The weight change:After the success of the model,the weight and the blank group were significantly decreased(P<0.01),The remaining groups were not significantly different from the normal group(P>0.05);The treatment groups gained weight more than before the treatment,The low dose group of the Shi Gan powder was statistically significant(P<0.05),The significance of the western medicine group,the middle dose and the high dose group of the Shi Gan powder were significant(P<0.01).②The changes of epileptic seizure latency:There was no significant difference between the groups in epileptic seizure latency before the treatment(P>0.05).After the treatment,the epileptic seizure latency of the model group decreased,the other groups had an increase in the incubation period.Among them,the incubation period of the western medicine group and the low,middle and high dose group of the Shi Gan powder was significantly increased.And it has significant means in statistics(P<0.01).③The changes of tonic convulsion latency:After the treatment in each group compared with before the treatment,the period of the tonic convulsion latency was significantly lower in the model group(P<0.05),while the other groups of rats increased their latency,the increase in the incubation period of low dose group of Shi Gan powder was statistically significant(P<0.05).The western medicine group,the middle dose and the high dose groups of the Shi Gan powder have significant significance(P<0.01).④The changes of tonic seizure rate:Compared with each groups before and after treatment,the model group had a significant increase in the tonic seizure rate(P<0.01),the rate of attack of the western medicine group,the middle and the high dose group of Shi Gan powder was significantly lower than before treatment(P<0.01),the low dose group of Shi Gan powder was not significantly different from the pretreatment(P>0.05).2.The observation of pathological changes of pentetrazol causes the hippocampal neurons in the epilepsy rats with Nissl dyeing:(1)Morphological change:Normal group of rat CA1 area neuron cone cell arrangement is closely ordered,the Neanderthal deep dye,the form regulation,the quantity is normal;In the model group,the pyramidal cells were shriveled,and the population was significantly reduced and disorganized,the Nissl cells are light and partially dissolved;The western medicine group neurons form,the cell shape is more regular,some of the neuron morphology is incomplete and shriveled,the amount is partially lost,the Nissl cells are hyperchromatic;The neurons in the low dose group of the Shi Gan powder decreased the number of cone cells in the group,and the Nissl cells were dissolving,Cone cells are loose and irregular;The number of cone cells in the middle dose of the Shi Gan powder was slightly reduced,the morphology was slightly shriveled,and Nissl cells’ staining was slightly shallow;The high-dose group of Shi Gan powder of neurons were arranged in an orderly manner,with a slightly reduced number of cells,and the Nissl body was deeply dyed.(2)The number of Nissl cells was compared:Compared with normal group,the number of the Nissl positive cells in the model group and the low dose group of Shi Gan powder was significantly lower,and it has statistically significant means(P<0.01);Compared to the model group,the normal group,the western medicine group,the middle dose and the high dose group of Shi Gan powder were significantly increased(P<0.01).There was a significant decrease in the number of Nissl positive cells in the model group and the low dose group of Shi Gan powder(P<0.01),the number of cells in the other groups was not very different from those:in the blank group,and the number of positive cells in the western medicine group ard the high dose group of Shi Gan powder were relatively high,and there was no statistically significant difference between the two groups(P>0.05).3.Expression of brain-derived neurotrophic factor BDNF in hippocampal neurons of PTZ rats after treatment(1)To compared with the blank group,the average grey density of BDNF was increased in the model group,the western medicine group,the low,medium and high dose group of Shi Gan powder,the model group:is the most significant(P<0.01),the average light density of BDNF was reduced in the model group,the western medicine group,the low,medium and high dose group of the Shi Gan powder(P<0.05),the model group is the most significant(P<0.01).(2)Compared with the model group,the AGD of the other treatment groups decreased,the western medicine group and the high dose group of the Shi Gan powder’ AGD decreased significantly(P<0.05),The low dose group of Shi Gan powder was not statistically significant(P>0.05);The AOD number of each group increased,the western medicine group and the high dose group Shi Gan powder’ AOD increased(P<0.05),The low dose of Shi Gan powder increment of the stone was not statistically significant(P>0.05).(3)Compare results between groups:The AGD of the model group and the low dose group of the Shi Gan powder was significantly increased by the other groups,the most significant of which was the model group(P<0.01),There was no significant difference between the blank group,the western medicine group,and the high dose group of Shi Gan powder(P>0.05);The AOD of the model group and the low dose group of Shi Gan powder were significantly lower than the other groups,with the most significant model group(P<0.01),there was no significant difference between the AOD in the blank group,the western medicine group,the middle and the high dose group of the Shi Gan powder(P>0.05).4.Effects of ion channel current amplitude:After the experiment,compared with the blank group,the model group was significantly elevated for the voltage dependence of the hippocampal neuron and the current amplitude(INaP)(P<0.05).Compared with the model group,the western medicine group and the low,middle and high dose groups of Shi Gan powder in the western medicine group reduced INaP,and the difference was statistically significant(P<0.05).However,the low,medium,high dose group of Shi Gan powder and the western drug group had no significant change in the voltage dependence of the transient Na + channel(INat),the statistical nonsense(P>0.05).Compared with the blank group,the model group the rat hippocampal neuron voltage and the transient K + channel(IA)decreased significantly(P<0.05).Compared with model group,western medicine group and Shi Gan scattered low,medium and high dose group can rise to reduce IA,the difference was statistically significant(P<0.05),but Shi Gan scattered low,medium and high dose group and western medicine group on the delayed rectifier K + channel(IK)had no significant change,there was no significant difference statistically(P>0.05)Conclution1.The low,medium,high dose group of Shi Gan powder and the sodium valproate were able to increase the weight of the preeclamine rats.2.The low,medium,high dose of Shi Gan powder,and sodium valproate could significantly prolong the incubation period of spasm of the pentium rats.Shi Gan dispersion,the high dose group and valproic acid sodium can prolong the incubation period of the rat tonic convulsion,and reduce the incidence of rat tonic convulsion,prompt Shi Gan scattered with valproate similar antiepileptic effect,and Shi Gan dispersion,high dose effect the most apparent.3.In Shi Gan powder and valproic acid sodium,high dose group were four nitrogen can increase the e model rat’s positive cells,improve its pathological form,prompt both have protective effect on neurons.4.Both high-dose and valproic acid in stone gans can reduce the average grey density of BDNF in the model,increase the average light density,It shows that the two have similar protective effects on hippocampal neurons.5.Shi Gan scattered low,medium and high dose group and valproic acid sodium could significantly lower e four nitrogen epilepsy caused by the voltage dependence of rat hippocampal neurons continuous Na + channel(INaP),current amplitude increase transient outward K + channels(IA),current amplitude of voltage dependent instantaneity Na + channel(INat)and delayed rectifier K + channel(IK)have no obvious effect on current,likely that Shi Gan scattered through the ion channel regulation play a role of anti epilepsy.6.Shi Gan scattered may by increasing the content of brain derived neurotrophic factor BDNF affect ion channel current amplitude,adjust the resting membrane potential,inhibitory synapses excitability,give play to the role of its anti epilepsy. |
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