Chinese Medicine Cai Tony Ended Epilepsy Soup Of Refractory Epilepsy In Rats Attack And Resistance Gene Expression

Background:Although a series of new antiepileptic drugs(AEDs)have been launched in the last two decades,drug-refractoriness remains a major problem.About 30%of patients are refractory to treatment using more than one AEDs,which is called medically intractable epilepsy.It has poor prognosis with increased morbidity and mortality.Therefore it is far-reaching important to investigate the drug resistance mechanism of medically intractable epilepsy,and develop nee strategies for treatment.Researchers found that many patients with tumor would yield resistance to drugs of chemotherapy.An important characteristic of pharmacoresistance is that most patients are resistant to one drug in the beginning,and it will be found that the other drugs will also be noneffective,even though these drugs act by different mechanisms.Which is called multidrug resistance(MDR).The following research indicate that membrane of cells of tumor express multidrug transporters,they act as drug efflux pump and restrict penetration of drug into cells,which produce chemotherapy-resistance in tumor.In the same way,there are the same pathophysiological mechanisms of drug resistance in medically intractable epilepsy.A series of researches concerning the relationship between multidrug transporters and drug resistance in medically intractable epilepsy have been done.The results suggest that several kinds of multidrug transporters are expressed in capillary endothelial cells and astrocytes of normal blood-brain barrier.There is accumulating ebidence demonstrating that multidrug transporters are over-expressed in capillary endothelial cells and astrocytes in epileptogenic brain tissue surgically resected from patients with medically intractable epilepsy. Multidrug transporters in the BBB are thought to act as an active defense mechanism,restricting the penetration of lipophilic substances into the brain. Over-expression of such transporters in epileptogenic tissue is thus likely to reduce the amount of drug that reaches the epileptic neurons,which would be a likely explanation for pharmacoresistance.P-glycoprotein(P-gp)and multidrug resistance-associated protein 1(MRP1)are candidates to cause AEDs resistance. They are ATP-binding cassette superfamily,and can accept a wide range of AEDs with different mechanisms as their potential substrates,and actively efflux drugs out of the brain.P-gp and MRP1 can be blocked by specific inhibitors which raises the option to use such inhibitors as adjunctive treatment for medically refractory epilepsy.Objective:In this study,In the base of building the animalmodel of refractory epilepsy by pilocarpine,we investigated P-gp/MDR1 mRNA and MRP1 mRNA influence by different medicine intervention.According to functional parmeter and pathohistology to analyses curative effect.Then the expression and distrbution of P-gp/MDR1 mRNA and MRP1 was research,localization in brain of rats with medically intractable epilepsy.We intended to study the role of P-gp/MDR1 mRNA and MRP1 mRNA on the the drug resistance mechanisms of medically intractable epilepsy,and investigate the effect on them by the herbs of complex prescription of chai bei zhi xian hot water.Materials and methods:We divided this research into three sections:In the first section,the model of medically intractable epilepsy was established by PILO,and it's praxiology was observed.5 group were used to excute this study:the normal control group(8 rats),the blank model group(20 rats), the herbs and western medicine group(20 rats),the herbs groups(rats),the western medicine group(20 rats).We observe the organism status of rats,the praxiology of rats,such as frequency,time length,averge time length,grade and defense response grade of epileptic seizure and body weight to analyses the curative effect.In the second section,We observed the rats brain tissue histomorphlogy by campeachy-eosin staining and Nissl's attaining.We investigate the change of neuron morphosis and the survivorship state of neuron and Nissl's body on the district of CA1,CA2,CA3 and dentate band of hippocampus,the district of frontal lobe and cortex of temporal lobe of cortical by campeachy-eosin staining and Nissl's attaining.In the third section,The expression of protein of P-gp and mRNA of MDR1 and MRP1 was investigeate in cortex and hippocampus of rats by the herbs of complex prescription of chai bei zhi xian hot water.We observed the expression and distrbution of P-gp/MDR1 mRNA and MRP1 in cortex and hippocampus of rats by Realtime PCR,immunohistochemical method and Western blotting.We intended to study the role of P-gp/MDR1 mRNA and MRP1 mRNA on the the drug resistance mechanisms of medically intractable epilepsy,and investigate the effect on them by the herbs of complex prescription of chai bei zhi xian hot water and carbamazepine intervention. Results:The results of first section:1 The change of seizur frequency of rats with intractable epilepsyCompared with that of the blank model group,the seizur frequency in the herbs and western medicine group,the herbs group and the western medicine group showed an obviously statistical decreasing(P＜0.05);Compared with that of the herbs and western medicine group,the seizur frequency in the herbs group showed an obviously statistical increasing(P＜0.05).2 The seizur time length of rats with intractable epilepsyCompared with that of the blank model group,the seizur time length in the herbs and western medicine group showed an obviously statistical decreasing(P＜0.05);Compared with that of the herbs and western medicine group,the seizur time length in the herbs group and the western medicine group showed an obviously statistical increasing(P＜0.05).3 The seizur averge time length of rats with intractable epilepsyAfter administer,compared with that of the blank model group,the seizur averge time length in the herbs and western medicine group,the herbs group and the western medicine group showed an obviously statistical decreasing(P＜0.05); Compared with that of the herbs and western medicine group,the seizur averge time length in the herbs group showed an obviously statistical increasing(P＜0.05).Compared with that of nulli-medicine,the seizur averge time length in the herbs and western medicine group showed an significant statistical decreasing(P＜0.01),in the herbs group and the western medicine group showed an obviously statistical decreasing(P＜0.05).4 The seizur grade of rats with intractable epilepsyAfter administer,compared with that of the blank model group,the seizur grade in the herbs and western medicine group and the western medicine group showed an obviously statistical decreasing(P＜0.05);Compared with that of nulli-medicine,the seizur grade in the herbs and western medicine group showed an significant statistical decreasing(P＜0.01),in the western medicine group showed an obviously statistical decreasing(P＜0.05).5 The defense response grade of rats with intractable epilepsyAfter administer,compared with that of the blank model group,the defense response grade in the herbs and western medicine group,the herbs group and the western medicine group showed an obviously statistical decreasing(P＜0.05); Compared with that of the herbs and western medicine group,the defense response grade in the western medicine group showed an obviously statistical increasing(P＜0.05).Compared with that of nulli-medicine,the defense response grade in the herbs and western medicine group and the herbs group showed an significant statistical decreasing(P＜0.01),in the western medicine group showed an obviously statistical decreasing(P＜0.05).6 The change of body weight of rats with all groupsAfter administer,compared with that of the normal control group,the body weight in the blank model group,the herbs and western medicine group,the herbs group and the western medicine group showed an obviously statistical decreasing(P＜0.05);Compared with that of nulli-medicine,compared with that of the normal control group,the body weight in the blank model group,the herbs and western medicine group,the herbs group and the western medicine group showed an obviously statistical decreasing(P＜0.05);Compared with that of the blank model group,the the body weight in the herbs and western medicine group showed an obviously statistical increasing(P＜0.05).The results of second section:1 The neuron showed obviously damage on the district of CA1,CA2,CA3 and dentate band of hippocampus,the district of frontal lobe and cortex of temporal lobe of cortical in the model of medically intractable epilepsy established by PILO.The most severe diseased region was the district of CA1 and CA3 of hippocampus,then is the district of CA2 and dentate band of hippocampus,the district of frontal lobe and cortex of temporal lobe of cortical.2 The herbs of complex prescription of chai bei zhi xian hot water and carbamazepine could relieve neuron damage on the district of CA1,CA2,CA3 and dentate band of hippocampus,the district of frontal lobe and cortex of temporal lobe of cortical in rats with intractable epilepsy,which hinted the two kinds of medinine could protect neuron.The results of third section:1 Compared with that of the normal control group,P-gp/MDR1 mRNA and MRP1 mRNA expression in cortex and hippocampus on rats with intractable epilepsy of the blank model group,the herbs and western medicine group,the herbs group and the western medicine group showed an obviously statistical increasing(P＜0.05);The amount of P-gp/MDR1 mRNA and MRP1 mRNA expression from high to low was by turns the western medicine group,the blank model group,the herbs group and the herbs and western medicine group.2 There was no obviously statistical difference between cortex and hippocampus about P-gp/MDR1 mRNA and MRP1 mRNA expression in all the groups.Conclusions:1 The manipulateion of the model of medically intractable epilepsy established by PILO was simple and it's achievement ratio and stability was fine.The curative effect of the way of the herbs combined western medicine is greatest than the others.2 The expression of P-gp/MDR1 mRNA and MRP1 mRNA are significantly increased in cortex and hippocampus of rats with medically intractable epilepsy.They are concerned with drug resistance of medically intractable epilepsy.3 The present study indicated that seizure may induce over-expression of P-gp/MDR1 mRNA and MRP1 mRNA in cortex and hippocampus of rats with medically intractable epilepsy,seizure and AED may be involved in up-regulation of P-gp/MDR1 mRNA and MRP1 mRNA expression.4 The herbs of complex prescription of chai bei zhi xian hot water may inhibit multidrug transporters,can decrease over-expression of P-gp/MDR1 mRNA and MRP1 mRNA.The one of mechanism on the herbs of complex prescription of chai bei zhi xian hot water healing refractory epilepsy may be it's inhibiting multidrug transporters,when it combining proper AED will make for the treatment of refractory epilepsy.