Study On The Mechanism Of Sodium Valproate Combined With Wuling Capsule In Treatment Of Epileptic Rats

ObjectiveThe study is based on the pentylenetetrazol epilepsy rats model by sodium valproate,Wuling capsule alone or integrated therapy,observing the behavior of the pentylenetetrazol epilepsy rats,studying the protective effect on hippocampal neurons by HE staining and the expressions of NMDAR1,GABRA2,GRM5,IL-1β,NF-?B / p65 and TLR4 in the hippocampus of epilepsy rats detected by immunohistochemically,real-time quantitative PCR method and Western Blot.MethodsSD rats both male and female were divided randomly into 5 groups: blank control group(group A),model group(group B),sodium valproate group(group C),Wuling capsule group(group D),sodium valproate combined with Wuling capsule group(group E).Each group had 10 rats and was given drugs by gavage treatment.Group A and group B were given normal saline;group C was given VPA;group D was given Wuling capsule;group E was given VPA and Wuling capsule.Given drugs at 9:00 every day,1h later,group A was injected with saline,and the other groups were injected PTZ solution,1 time a day,4 consecutive weeks.Observing and recording the level of epileptic seizure in half an hour after the injection of PTZ solution.After 4 weeks of treatment,5 rats were randomly selected in each group,anesthetized by 10% chloral hydrate intraperitoneal injection,then perfused with saline and 4% formaldehyde and removed the brain tissues to prepare paraffin sections to perform the pathological examination and used immunohistochemically to detect the expressions of NMDAR1,GABRA2,GRM5,IL-1β,NF-?B / p65 and TLR4 in the hippocampus of epilepsy rats.The last rats were randomly selected in each group,anesthetized by 10% chloral hydrate intraperitoneal injection,then perfused with saline and 4% formaldehyde and removed the hippocampus,used real-time quantitative PCR method and Western Blot to detect the expressions of NMDAR1,GABRA2,GRM5,IL-1β,NF-?B / p65 and TLR4 in the hippocampus of epilepsy rats.1.In the model of epilepsy induced by PTZ,sodium valproate combined with Wuling capsule group could suppress epilepsy seizures,significantly reduced the level and total number of seizures,and had better effect than sodium valproate group,Wuling capsule group.2.Sodium valproate group,Wuling capsule group,sodium valproate combined with Wuling capsule group all could improve the damage of neurons in hippocampal CA3 area,sodium valproate group and sodium valproate combined with Wuling capsule group were better than Wuling capsule group.3.The result of immunohistochemically: NMDAR1、GRM5、NF-?B/p65、TLR4 and IL-1β in hippocampus of rats in the blank control group,the expression was less;in the model group,there was a lot of expression;in the sodium valproate group,Wuling capsule group,sodium valproate combined with Wuling capsule group,the expression decreased significantly(P<0.05).However,IL-1β in hippocampus of rats has no obvious difference(P>0.05).The expression of GABRA2 in hippocampus of rats in the blank control group was large;in the model group,the expression was less;in the sodium valproate group,Wuling capsule group,sodium valproate combined with Wuling capsule group,the expression increased significantly(P<0.05).4.The result of real-time quantitative PCR method: NMDAR1、GRM5、NF-?B/p65、TLR4 and IL-1β in hippocampus of rats in the blank control group,the expression was less;in the model group,there was a lot of expression;in the sodium valproate group,Wuling capsule group,sodium valproate combined with Wuling capsule group,and the expression trend decreased significantly.The expression of GABRA2 in hippocampus of rats in the blank control group was large;in the model group,the expression was less;in the sodium valproate group,Wuling capsule group,sodium valproate combined with Wuling capsule group,the expression had ricing trend.5.The result of Western Blot : NMDAR1、GRM5、NF-?B/p65、TLR4 and IL-1β in hippocampus of rats in the blank control group,the expression was less;in the model group,there was a lot of expression;in the sodium valproate group,Wuling capsule group,sodium valproate combined with Wuling capsule group,the expression decreased significantly(P<0.05).The expression of GABRA2 in hippocampus of rats in the blank control group was large;in the model group,the expression was less;in the sodium valproate group,Wuling capsule group,sodium valproate combined with Wuling capsule group,the expression increased significantly(P<0.05).ConclusionFrom the results,sodium valproate combined with Wuling capsule could better control the seizure of epileptic rats,reduced the damage of hippocampal neurons,the mechanism could be related of through GABA/GLU signaling pathway and TLR4/NF-?B signaling pathway,which Results were related,to increase the expression of NMDAR1、GRM5、NF-?B/p65、TLR4and IL-1β,and to reduce the expression of GABRA2.