To Investigate The Effects Of Huazhuo Jiedu Shugan Decoction On Learning And Memory In Epilepsy Model Rats Via CAMP-PKA-CREB Pathway

Objective: To observe the changes of behavior,pathological morphology and Cyclic adenosine monophosphate-protein kinase A-cyclic adenosine monophosphate effector binding protein(cAMP-PKA-CREB)pathway protein expression in epileptic rats treated with Huazhuo Jiedu Shugan decoction(HJSD),and to investigate the effect of learning and memory ability in epileptic model and its possible mechanism.Methods: Sixty SPF grade SD rats were randomly divided into normal group,model group,western medicine group,Huazhuo Jiedu Shugan decoction low dose group(HJSD-L),Huazhuo Jiedu Shugan decoction medium dose group(HJSD-M)and Huazhuo Jiedu Shugan decoction high dose group(HJSD-H).The model of epileptic rats was established by pentylenetetrazol chemical ignition,and the epileptic seizures and behavior of the rats were observed.After the epilepsy model was established,HJSD and valproate acid were given for grouping treatment.During the treatment cycle,changes in epileptic seizures behavior were observed and recorded in each group.After the treatment,the learning and memory ability of the rats was evaluated by cognitive behavioral experiments,the pathomorphological changes of hippocampal neurons in rats were observed by optical microscope,and the expression of cAMP-PKA-CREB pathway related protein immunoreactive cells in the hippocampus of the rats was observed by immunohistochemistry.All experimental data were recorded and analyzed by medical statistics.Results: 1.General observation: rats in the normal group grew well and gained weight rapidly.At the end of modeling,compared with the normal group,the epileptic model rats showed significant growth retardation,weight loss(P<0.01),hair removal and poor luster.After drug treatment,the diet and water consumption of the rats in each group increased,and the indumentum was improved and the depilation was alleviated,but the body weight was still significantly less than that of the normal group(P<0.01).The weight gain of rats in western medicine group,HJSD-M group and HJSD-H group after treatment was significantly higher than that in model group(P<0.01).2.Seizure status: at the 21 st day of treatment,compared with the model group,the seizure grade of the western medicine group and the HJSD-H group were significantly lower and the seizure latency were significantly prolonged(P<0.05).At the 28 th day of treatment,the seizure grade in HJSD-M group was lower and the seizure latency was longer than that in the model group(P<0.05).Compared with the model group,there were significant differences between the western medicine group and the HJSD-H group(P<0.01).3.Morris Water Maze test:(1)Place navigation: there was no significant difference between each group in the first two days of the experiment(P>0.05),and the escape latency of rats in the normal group was significantly less than that in the model group in the following two days(P<0.05).On the 4th day of the experiment,the escape latency of rats in the HJSD-M group was significantly shorter than that in the model group(P<0.05),while the escape latency of rats in the western medicine group and the HJSD-H group was significantly different from that in the model group(P<0.01).(2)Spatial probe test: The number of rats crossing the platform in the model group was significantly less than that in the normal group(P<0.01).Compared with the model group,the number of rats crossing the platform in the HJSD-M group was significantly higher(P<0.05),while there were significant differences between the western medicine group and the HJSD-H group(P<0.01).4.Radial Arm Maze test: Compared with the normal group,the values of working memory error(WME),reference memory error(RME)and total error(TE)in the model group were significantly increased(P<0.01).Compared with the model group,the values of WME,RME and TE in the western medicine group and the HJSD-H group were significantly lower(P<0.05,P<0.01),while the values of RME and TE in the HJSD-M group were significantly lower(P<0.05,P<0.01).5.Pathomorphological observation: the hippocampal nerve cells of rats in the normal group were abundant,arranged in a regular structure and had distinct levels.In the model group,the number of nerve cells decreased sharply,and the nerve cells were arranged in disorder,deforming and necrosis,cell nucleus shrinkage,and there were more apoptotic bodies,glial cells and inflammatory cells.After treatment,the nerve cells in the western medicine group and the HJSD group were arranged in a orderly manner,with slight nuclear shrinkage,a small amount of apoptotic corpuscule,glial cells and inflammatory cells.6.Immunohistochemical detection: the expression levels of cAMP-PKA-CREB pathway related protein immunoreactive cells in the hippocampus of the normal group,western medicine group,HJSD-M group and HJSD-H group were significantly higher than those in the model group(P<0.05,P<0.01).Conclusion: 1.The model of epileptic rats can be successfully induced by pentylenetetrazol chemical ignition,and the model is stable and reliable.The epilepsy model rats showed significant learning and memory disorders.2.HJSD has significant antiepileptic effect,and can significantly improve the learning and memory ability of epileptic rats.3.The improvement of learning and memory ability in epileptic rats by HJSD may be caused by the up-regulation of the signal of the cAMP-PKA-CREB pathway and the improvement of the expression level of pathway-related proteins,thus resulting in the improvement of the learning and memory behavior.