The Clinical Study Of Chaibei Zhixian Decoction In The Treatment Of Drug-resistant Epilepsy And The Effect Of Its Blood Components On The Concentration Of Carbamazepine In The Brain Tissue Of KA-induced Epilepsy Rats

Drug resistant epilepsy has become the primary problem of epilepsy treatment since it is insensitive to antiepileptic drugs(AEDs).Recently,more and more new AEDs are developed but there are fewer breakthroughs on improving drug resistant epilepsy,which may be related with the failure to regulate patient's potential resistance mechanisms.Long-term clinical practices show that adding Chinese medicine in AEDs can improve antiepileptic effect.The Chaibei Zhixian Decoction which based on the TCM syndrome research of drug resistant epilepsy develops from two classic ancient formulas and shows remarkable effect in controlling seizures.To evaluate the clinical efficacy and safety of Chaibei Zhixian Decoction adding AEDs in improving drug resistant epilepsy is meaningful for investigating the optimized treatment scheme of drug resistant epilepsy.Further analysis the regulation of the compound and its main components in plasma in resistance proteins and genes will help us to clear the anti-drug resistant epilepsy mechanism of Chinese medical.[Objectives]To evaluate the effect and safety of Chaibei Zhixian Decoction adding AEDs in treating drug resistant epilepsy by randomized control trial;to observe the influence of Chaibei Zhixian Decoction and its main components in plasma like Gastrodin,Gastrodigenin and Vanillin on the expression of P-glycoprotein(P-gp)and Multi-Drug Resistance Genel(Mdrl)in rat's hippocampus as well as the concentrations of carbamazepine(CBZ)and its anticonvulsant active metabolite Carbamazepine 10,11-Epoxide(CBZE)in rat's whole brain tissue by the animal experiment.[Methods]Clinical trial:The patients with drug resistant epilepsy focal seizure accompanied by disturbance of consciousness were included and randomly divided into treatment group and control group.Patients from the treatment group were given original AEDs together with Chaibei Zhixin granules and patients from the control group were given original AEDs together with placebo granules.The treatment course was 3 months.The seizure frequency,50%response rate(RR),freedom rate and Liverpool Seizure Severity Scale(LSSS)score were used to assess the improvement of seizures;the Neurological Disorders Depression Inventory for Epilepsy(NDDI-E)score,the Hamilton Depression Scale(HAMD)-24 score and Hamilton Anxiety Scale(HAMA)score were used to evaluate the improvement of depression and anxiety symptoms.The Quality of Life in Epilepsy(QOLIE)-89 score was selected to evaluate the improvement of life quality in patients.The rate of adverse events include Blood routine,liver and kidney function,other abnormal symptoms and signs was selected to evaluate safety of the combined treatment.Animal experiment:The drug resistant epilepsy rat models were prepared by injecting kainic acid(KA)into the lateral ventricle.And the successful model rats were randomly divided into model group,Carbamazepine group,Chaibei Zhixian Decoction +Carbamazepine group,Gastrodin+ Carbamazepine group,Gastrodigenin +Carbamazepine group,Vanillin + Carbamazepine group,and set a blank group.The course of gavage treatment was 60 days.The expressions of P-gp and Mdr1a/b mRNA in the hippocampus of rats were detected by Western Blot and RT-PCR.And the concentrations of CBZ and CBZE in the whole brain tissue were detected by Liquid chromatography-mass spectrometry(LC-MS).[Results]Clinical trial1.Basic data:A total of 67 patients were included with 34 in treatment group and 33 in control group.There were statistical differences in gender and current application of AEDs(P<0.01,P<0.05)while no statistical differences in other basic clinical data.2.Situation of Seizures:(1)seizure frequency:The seizure frequency of treatment group was lower than that of the control group after 3 months treatment(P<0.05);the reduction of seizure frequency in the treatment group was higher than the control group after treated for 1,2,and 3 months(P<0.05).(2)50%RR:The 50%RR of treatment group was higher than the control group after treated for 2 and 3 months(P<0.05,P<0.01).(3)Freedom rate:The freedom rate of treatment group was higher than the control group after treated for 3 months(P<0.05).(4)LSSS score:The LSSS score of treatment group was lower than the control group(P<0.05)and the reduction of LSSS score in treatment group was larger than the control group after 3 months treatment(P<0.01).3.Accompanied depression symptoms:(1)Number of patients with depression evaluated by NDDI-E and HAMD scales:The number of patients with depression evaluated by NDDI-E and HAMD scales showed no statistical differences between the two groups after treated for 3 months.HAMD score:There was no statistical difference on HAMD score between the two groups after treated for 3 months while the reduction of HAMD score in treatment group was larger than the control group(P<0.01).(2)The results of NDDI-E and HAMD scales showed medium consistency without statistical differences,but this comparison result is related to that whether 8?HAMD<20 was evaluated as "depression".4.Accompanied anxiety symptoms:Number of patients with anxiety evaluated by HAMA scale:There was no statistical difference on the number of patients with anxiety evaluated by HAMA scale between the two groups after 3 months treatment.HAMA score:The HAMA score showed no statistical difference between the two groups after treated for 3 months while the HAMA score reduction of treatment group was larger than the control group(P<0.01).5.Life quality:QOLIE-89 total score:There was no statistical difference between on the QOLIE-89 total score the two group after 3 months treatment but the increase of QOLIE-89 total score in the treatment group was higher than the control group(P<0.01).Score for single item:The scores for 3 single items and 1 clause of the treatment group were higher than the control group after 3 months treatment(energy/fatigue,attention/concentration,medication effects,change in health,P<0.05).The score increase amplitudes of 6 single items and 2 clauses of treatment group were larger than the control group(health perceptions,work/drive/social function,emotional well-being,seizure worry,P<0.05;pain,energy/fatigue,change in health,over health,P<0.01).6.Safety:There were no adverse events relating to the treatment of adding Chaibei Zhixian Decoction.Animal experiment1.Expression of P-gp in hippocampus:compared with the blank group,the P-gp expression of hippocampus in model and Carbamazepine group increased(P<0.01).Compared with the model group,the P-gp expression of hippocampus in the Carbamazepine group increased(P>0.05),but the P-gp expression of hippocampus in the Chaibei Zhixian Decoction + Chaibei Zhixian Decoction group decreased(P<0.05).Compared with the Carbamazepine group,that of Chaibei Zhixian Decoction+Chaibei Zhixian Decoction group,Gastrodin+ Chaibei Zhixian Decoction group,Gastrodigenin + Carbamazepine group decreased(P<0.01,P<0.05,P<0.05).Compared with the Chaibei Zhixian Decoction+Chaibei Zhixian Decoction group,that of Vanillin + Carbamazepine group increased(P<0.05).2.Expression of Mdrla/b mRNA in hippocampus:Compared with the blank group,the expression of Mdrla mRNA in model group increased without statistical difference and expression of Mdrlb mRNA in model group increased(P<0.05),expression of Mdrla mRNA and Mdrlb mRNA in Carbamazepine group increased(P<0.01,P<0.05).Compared with the model group,expression of Mdrla mRNA and Mdrlb mRNA in Carbamazepine group increased without statistical difference.Compared with Carbamazepine group,expression of Mdr1a mRNA and Mdr1 b mRNA in Chaibei Zhixan Decoction+Carbamazepine group and Gastroigenin+ Carbamazepine group decreased(P<0.01,P<0.05,P<0.05).Compared with Chaibei Zhixan Decoction+Carbamazepine group,expression of Mdrla mRNA and Mdrlb mRNA in Gastrodin+Carbamazepine group increased(P<0.05),expression of Mdrlb mRNA in Vanillin+ Carbamazepine group increased(P<0.05).Compared with Gastroigenin +Carbamazepine group,expression of Mdrlb mRNA in Vanillin+ Carbamazepine group increased(P<0.05).3.Cerebral CBZ and CBZE concentrations:CBZ concentration:There were no statistical differences among the five groups.CBZE concentration:Compared with Carbamazepine group,CBZE concentration of Chaibei Zhixan Decoction?Carbamazepine group and Gastroigenin+Carbamazepine group increased(P<0.05).Compared with Chaibei Zhixian Decoction + Carbamazepine group,CBZE concentration of Gastrodin +Carbamazepine group decreased(P<0.05).For both cerebral CBZ and CBZE concentration,the concentration orders from highest to lowest were Chaibei Zhixian Decoction +Carbamazepine group,Gastrodigenin + Carbamazepine group,Vanillin + Carbamazepine group,Carbamazepine group,Gastrodin+Carbamazepine group[Conclusions]1.Treating patients with drug resistant epilepsy focal seizure accompanied by disturbance of consciousness by Chaibei Zhixian Decoction adding AEDs can reduce the seizure frequency,decrease the severity,improve the symptoms of accompanied depression and anxious so as to improve the life quality.The clinical application of Chaibei Zhixian Decoction adding AEDs is safe and effective.2.Chaibei Zhixian Decoction and Gastrodigenin can increase the CBZE concentration in the brain of kainic acid-induced epilepsy rats and down-regulation of P-gp,Mdrla/b mRNA expression in hippocampus may be one of the possible mechanisms.