Study On Mechanism Of Songling Xuemaikang Capsules Combined With CBZ In The Treatment Of Kainic Acid Refractory Epileptic Rats

ObjectiveThis study is based on the kainic acid refractory epilepsy rats model by the Songling Xuemaikang Capsules,Carbamazepine alone or integrated therapy,observing the kainic acid refractory epilepsy rats' behavioral changes and situation of seizure.To study the effect of Songling Xuemaikang Capsules on learning and memory ability of epilepsy rats by Morris water maze.To study the effect of Songling Xuemaikang Capsules on hippocampal neurons by Nissl.To study the expression of p-Akt?Akt?Caspase-9?P-GP?MRP1 in hippocampus by Western Blot.Methods1?70 SD male rats were divided randomly into 5 groups: blank control group,model group,Carbamazepine group,Songling Xuemaikang Capsules group and Songling Xuemaikang Capsules combined with Carbamazepine group.Each group was further divided into 2 sub-groups: Administration of 30 d group and 60 d group.Blank control group was received saline,the other groups were injected kainic acid in lateral ventr icle.Blank control group and model group were treated with normal saline.The other groups were given the corresponding drugs.During the experiment,the symptoms changes and the frequency of the spontaneous recurrent seizures were recorded.2?70 rats were selected from 30 d and 60 d sub-groups(7 rats in each group)to perform Morris water maze test.The escape latency and the swimming frequency of crossing the platform were recorded.3?30 rats selected randomly from 30 d and 60 d sub-groups(3 rats in each group)were anesthetized by 10% chloral hydrate intraperitoneal injection,then perfused with saline and 4% formaldehyde.Nissl and HE staining were performed.4?40 rats selected randomly from 30 d and 60 d sub-groups(4 rats in each group)were anesthetized by 10% chloral hydrate intraperitoneal injection.The hippocampal tissue was removed immediately.Western blot was used to detect the expression of p-Akt?Akt?Caspase-9?P-GP?MRP1 of each group.Results1?Comparing with model group,the stage of seizures and SRS frequency were decreased in Songling Xuemaikang Capsules group,but there was no significant difference(P>0.05).With the extention of treatment time,the stage of seizures was decreased in Carbamazepine group and there was no signif icant difference(P>0.05).The stage of seizures and SRS frequency in Songling Xuemaikang Capsules combined with Carbamazepine group were observably decreased(P<0.05).Comparing with Carbamazepine group,SRS frequency was markedly decreased(P<0.05).2?In place navigation test,with the increased of the number of training days,the latency of each group were gradually decreased and model group was significantly longer than the blank control group on the same day(P<0.05,P<0.01).Comparing with model group,the latency of carbamazepine group had been shortened,but there was no significant difference(P>0.05).The latency of Songling Xuemaikang Capsules group and the combined group was signif icantly shortened(P<0.05,P<0.01).In spatial probe test,comparing with the model group,the times of crossing platform in each group were increased.The times of crossing platform were observably increased in Songling Xuemaikang Capsules group and the combined group(P<0.05 or P<0.01).Carbamazepine could not observably increased the times of crossing platform(P>0.05).3?With the extention of treatment time,comparing with the model group,the number of neurons in rats' hippocampal CA3 area was increased in Songling Xuemaikang Capsules group,but there was no significant difference(P>0.05).The number of neurons in rats were obviously increased in Carbamazepine group and combined group(P<0.01).The combined group has a larger number of neurons in hippocampal CA3 area than Carbamazepine group(P<0.05).4?With the extention of treatment time,comparing with the model group,Carbamazepine and Songling Xuemaikang Capsules alone could not markedly increase the expression of p-Akt(P>0.05)and Songling Xuemaikang Capsules could not evidently decrease the expression of Caspase-9(P>0.05).Songling Xuemaikang Capsules combined with Carbamazepine could increase the expression of p-Akt and inhibit the expression of Caspase-9,while the effect of combined group was more efficient than Songling Xuemaikang Capsules group and Carbamazepine group.Therefore,while taking Carbamazepine to control epilepsy,integrated Songling Xuemaikang Capsules can inhibit apoptosis better.5?Comparing with Carbamazepine group,the P-GP expression in hippocampus of rats of combined group was signif icantly reduced(P<0.01),MRP1 expressionin hippocampus of rats was decreased,but there was no significant difference(P>0.05).It showed that Songling Xuemaikang Capsules combined with Carbamazepine could weaken Carbamazepine' induction of P-GP,but had no significant effect on MRP1.ConclusionSongling Xuemaikang Capsules combined with Carbamazepine could more efficiently control the seizures of kainic acid refractory epilepsy rats,reduce the damage of hippocampal neurons,improve their learning and memory ability.The mechanism may be related to enhancing the expression of p-Akt,inhibiting the expression of Caspase-9 and decreasing the expression of P-GP.