Observation On The Therapeutic Effect Of Chibai Zhixu Decoction In Treating Refractory Epilepsy And The Mechanism Of Intervention Of P - GP

It has long been known that epilepsy is one of the most common chronic brain disorder of nervous system. About 65 million people worldwide have epilepsy. Epilepsy can lead to a lot of adverse effects, such as accidental injury, neurological or psychiatric disorders comorbidities, social dysfunction, the lower quality of life, and even death. High mortality in epilepsy patients is mostly due to brain injury, status epilepticus, suicide, pneumonia and sudden death. Nearly 30% of the epilepsy patients receiving anti-epileptic drugs (AEDs) still have seizures and most of them will evolve in to intractable epilepsy (IE).In recent years, a large number of experimental studies have shown that one of the most important resistance mechanism of IE may be the high expression of multidrug resistance proteins, especially p-glycoprotein(P-gp).The inhibitions of multidrug resistance proteins expression increasing the concentration of AEDs in ehlesions brain tissue are becoming more and more popular in clinical domain.Treating epilepsy with Traditional Chinese Medicine(TCM) has a long history. A large number of clinical studies have shown that treating epilepsy with TCM has reliable clinical curative effect, and Chinese herbs bring less adverse reaction.Chaibei Zhixian Decoction combined with AEDs treatment of intractable epilepsy can inhibit high expression of multidrug resistance proteins,so as to pretect the brain. Chaibei Zhixian Decoction combined with AEDs has a definite therapeutic effect on Intractable Epilepsy.Objective The aim of this study was to investigate the clinical efficacy and safety of Chaibei Zhixian Decoction as add-on therapy in the treatment of Intractable Epilepsy; and to observe the regulated effects of Chaibei Zhixian Decoction on the expressions and the mechanisms of P-gp in rat brain and blood brain barrier.Methods Clinical research uses the method of matched case-control study, and there are respectively 70 patients with intractable epilepsy in the treatment group and control group Each patient in the control group is treated with only Antiepileptic Drugs (AEDs), and that of the treatment group is treated with Chaibei Zhixian Decoction with AEDs. The primary outcome measure are the total effective rate, scores of seizure severity and the accompanying symptoms of epilepsy attack symptoms, abnormal degree of EEG.Record retention rate of drugs and adverse reactions.The epilepsy model was reproduced by kainic acid which was injected into the cella lateralis of rats.The model rats were randomly divided into 6 groups as following:normal group, sham group, model group, Carbamazepine group, Chinese Medicine group, and Carbamazepine combined with Chinese Medicine group. After 3 months treatment, Western blot is used to assess the expression of COX-2and NF-κB in hippocampus and Real-time PCR is used to detect the amplifications of COX-2 mRNA and NF-κB mRNA. Rat Brain Microvascular Endothelial Cells(RBMEC)were exposed to L-glutamate to up-regulate the expression of P-gp,which were up-regualte as multi-drug resistant epilepsy. After treatments, the solutions were replaced with a concentrationof Chaibei Zhixian Decoction. Western blot is used to assess the expression of COX-2、NF-κB and P-gp, and Elisa is used assess the expression of COX-2 and PGE2.Results (1) The treatment group was superior to the control group in decreasing the seizure frequency(P<0.05),scores of seizure severity (P<0.05),and scores of the accompanying symptoms of epilepsy attack symptoms(P<0.05), without any obvious adverse reactions; (2)Injection kainic acid in the cella lateralis of rats can up-regulate the level COX-2and NF-κB. Treatment with Chaibei Zhixian Decoction can decrease the expression of COX-2, NF-κB and COX-2 mRNA、NF-κB mRNA(P<0.05);(3)Rat Brain Microvascular Endothelial Cells(RBMEC) exposed to L-glutamate can up-ragulate the expression of P-gp. The COX-2、 NF-κB and P-gp expression of a concentration (100 ug/mL、250 ug/mL、500 ug/mL)of Chaibei Zhixian Decoction group is significantly lower than that of model cell group(P< 0.05); The COX-2 expression of a concentration (100 ug/mL,250 ug/mL,500 ug/mL)of Chaibei Zhixian Decoction group is significantly lower than that of model cell group in the cell culture supernatant(P<0.05); The PGE2 expression of a concentration (500 ug/mL)of Chaibei Zhixian Decoction group is significant lower than that of model cell group in the cell culture supernatant(P<0.05).Conclusions Chaibei Zhixian Decoction combined with AEDs has a definite therapeutic effect on Intractable Epilepsy, without obvious adverse reaction. Chaibei Zhixian Decoction decreases the expression of COX-2、NF-κB and COX-2 mRNA、NF-κB mRNA in rat, and it decreases the expression of COX-2、PGE2、NF-κB, P-gp in RBMEC. Chaibei Zhixian Decoction can decrease the expression of P-gp by decreasing the expression of COX-2、PGE2 and NF-κB, so as to increase the AEDs in the brain, then it can reverse the multidrug resistant of Intractable Epilepsy.