苯妥英钠耐药性颞叶内侧癫痫大鼠模型的构建
|
摘要
目的验证锂-匹罗卡品化学诱导及抗癫痫药物(AEDs)筛选构建的苯妥英钠耐药性颞叶内侧癫痫大鼠模型脑中AEDs浓度及多药转运蛋白表达。方法实验选取雌性6～8周SD大鼠30只,体重160～180 g(广东省实验动物研究所),采用锂-匹罗卡品化学诱导大鼠发生慢性颞叶内侧癫痫,视频脑电图(VEEG)监测苯妥英钠筛选期治疗效果,将颞叶内侧癫痫大鼠分成耐药性及不耐药性模型。采用活体微透析技术验证耐药性及不耐药性模型鼠脑中AEDs的浓度差异,免疫组织化学方法检测两组模型鼠脑中P糖蛋白的表达。结果有16只大鼠成功构建为慢性颞叶内侧癫痫模型,筛选出耐药模型大鼠6只(6/16)。苯妥英钠耐药性模型大鼠脑/血浆的时间药物浓度曲线的曲线下面积比值显著低于不耐药性模型大鼠(0.15±0.03 vs. 0.28±0.05,P<0.05)。苯妥英钠耐药性模型鼠较不耐药性模型鼠海马各区P糖蛋白表达明显增高(P<0.05)。结论苯妥英钠耐药性颞叶内侧癫痫大鼠脑中AEDs浓度低,可能与脑中P糖蛋白过表达有关。
Objectives The purpose of this study is to verify the phenytoin-resistant mesial temporal lobe epilepsy(MTLE) induced by Li-pilocarpine and screened by antiepilepsy drug(AEDs). Methods The rats with MTLE were induced by Li-pilocarpine, which were screened by effect of phenytoin treatment monitored by vedio-EEG. The living microdialysis technology was used for verification of drug concentration in brain of drug-resistant and drugresponsive rat model, and the P-glycoprotein expression was detected by immunohistochemical method. Results Sixteen rats with chronic MTLE were successfully induced in total 30 rats, among which, 6 drug-resistant rats with MTLE were screened. The brain/plasma ratio of area under the curve in drug-resistant rats was significantly lower than that of drugresponsive rats(0.15±0.03 vs. 0.28±0.05, P<0.05). In addition, the P-glycoprotein expression in brain of drug-responsive rats was obviously higher than that of drug-responsive rats(P<0.05). Conclusions The low concentration of phenytoin in drug-resistant rat model with MTLE was verified that might be related to the over-expressed P-glycoprotein in brain.
Objectives The purpose of this study is to verify the phenytoin-resistant mesial temporal lobe epilepsy(MTLE) induced by Li-pilocarpine and screened by antiepilepsy drug(AEDs). Methods The rats with MTLE were induced by Li-pilocarpine, which were screened by effect of phenytoin treatment monitored by vedio-EEG. The living microdialysis technology was used for verification of drug concentration in brain of drug-resistant and drugresponsive rat model, and the P-glycoprotein expression was detected by immunohistochemical method. Results Sixteen rats with chronic MTLE were successfully induced in total 30 rats, among which, 6 drug-resistant rats with MTLE were screened. The brain/plasma ratio of area under the curve in drug-resistant rats was significantly lower than that of drugresponsive rats(0.15±0.03 vs. 0.28±0.05, P<0.05). In addition, the P-glycoprotein expression in brain of drug-responsive rats was obviously higher than that of drug-responsive rats(P<0.05). Conclusions The low concentration of phenytoin in drug-resistant rat model with MTLE was verified that might be related to the over-expressed P-glycoprotein in brain.
引文
1Kwan P and Brodie MJ. Definition of refractory epilepsy:defining the indefinable?. Lancet Neurol, 2010, 9(1):27-29.
2Golyala A and Kwan P. Drug development for refractory epilepsy:The past 25 years and beyond. Seizure, 2017, 44:147-156.
3Santulli L, Coppola A, Balestrini S, et al. The challenges of treating epilepsy with 25 antiepileptic drugs. Pharmacol Res, 2016, 107:211-219.
4Banerjee Dixit A, Sharma D, Srivastava A, et al. Upregulation of breast cancer resistance protein and major vault protein in drug resistant epilepsy. Seizure, 2017, 47:9-12.
5Hartz AM, Pekcec A, Soldner EL, et al. P-gp Protein Expression and Transport Activity in Rodent Seizure Models and Human Epilepsy. Mol Pharm, 2017, 14(4):999-1011.
6Feldmann M and Koepp M. ABC Transporters and Drug Resistance in Patients with Epilepsy. Curr Pharm Des, 2016,22(38):5793-5807.
7Volk HA, Loscher W. Multidrug resistance in epilepsy:rats with drug-resistant seizures exhibit enhanced brain expression of Pglycoprotein compared with rats with drug-responsive seizures.Brain, 2005, 128(Pt 6):1358-1368.
8方子妍,郭彩凤,吴逢春,等.普朗尼克P85修饰的苯妥英钠纳米粒对颞叶内侧癫痫大鼠模型的脑靶向作用.中国神经精神疾病杂志, 2017, 43(06):356-361.
9方子妍,郭彩凤,吴逢春,等. Tariquidar提高苯妥英钠在颞叶内侧癫痫模型鼠脑中的分布.临床医学工程, 2017, 24(06):762-764.
10Fang ZY, Chen SD, Chen YS, et al. Pluronic P85 enhances the delivery of phenytoin to the brain versus verapamil in vivo. Latin American Journal of Pharmacy, 2014, 33(5):812-818.
11Fang Z, Chen S, Qin J, et al. Pluronic P85-coated poly(butylcyanoacrylate)nanoparticles overcome phenytoin resistance in P-glycoprotein overexpressing rats with lithiumpilocarpine-induced chronic temporal lobe epilepsy. Biomaterials,2016, 97:110-121.
12Racine RJ. Modification of seizure activity by electrical stimulation.II. Motor seizure. Electroencephalogr Clin Neurophysiol, 1972,32(3):281-294.
13Loscher W. Animal Models of seizures and epilepsy:past, present,and future role for the discovery of antiseizure drugs. Neurochem Res, 2017, 42(7):1873-1888.
14刘阳,韩春雷,孟凡刚.匹罗卡品癫痫模型研究进展.中华神经外科杂志, 2014, 30(1):88-90.
15 陈树达,陈子怡,方子妍,等.普朗尼克P85与维拉帕米对苯妥英钠脑靶向分布的比较.中山大学学报(医学科学版), 2014, 35(2):161-168.
2Golyala A and Kwan P. Drug development for refractory epilepsy:The past 25 years and beyond. Seizure, 2017, 44:147-156.
3Santulli L, Coppola A, Balestrini S, et al. The challenges of treating epilepsy with 25 antiepileptic drugs. Pharmacol Res, 2016, 107:211-219.
4Banerjee Dixit A, Sharma D, Srivastava A, et al. Upregulation of breast cancer resistance protein and major vault protein in drug resistant epilepsy. Seizure, 2017, 47:9-12.
5Hartz AM, Pekcec A, Soldner EL, et al. P-gp Protein Expression and Transport Activity in Rodent Seizure Models and Human Epilepsy. Mol Pharm, 2017, 14(4):999-1011.
6Feldmann M and Koepp M. ABC Transporters and Drug Resistance in Patients with Epilepsy. Curr Pharm Des, 2016,22(38):5793-5807.
7Volk HA, Loscher W. Multidrug resistance in epilepsy:rats with drug-resistant seizures exhibit enhanced brain expression of Pglycoprotein compared with rats with drug-responsive seizures.Brain, 2005, 128(Pt 6):1358-1368.
8方子妍,郭彩凤,吴逢春,等.普朗尼克P85修饰的苯妥英钠纳米粒对颞叶内侧癫痫大鼠模型的脑靶向作用.中国神经精神疾病杂志, 2017, 43(06):356-361.
9方子妍,郭彩凤,吴逢春,等. Tariquidar提高苯妥英钠在颞叶内侧癫痫模型鼠脑中的分布.临床医学工程, 2017, 24(06):762-764.
10Fang ZY, Chen SD, Chen YS, et al. Pluronic P85 enhances the delivery of phenytoin to the brain versus verapamil in vivo. Latin American Journal of Pharmacy, 2014, 33(5):812-818.
11Fang Z, Chen S, Qin J, et al. Pluronic P85-coated poly(butylcyanoacrylate)nanoparticles overcome phenytoin resistance in P-glycoprotein overexpressing rats with lithiumpilocarpine-induced chronic temporal lobe epilepsy. Biomaterials,2016, 97:110-121.
12Racine RJ. Modification of seizure activity by electrical stimulation.II. Motor seizure. Electroencephalogr Clin Neurophysiol, 1972,32(3):281-294.
13Loscher W. Animal Models of seizures and epilepsy:past, present,and future role for the discovery of antiseizure drugs. Neurochem Res, 2017, 42(7):1873-1888.
14刘阳,韩春雷,孟凡刚.匹罗卡品癫痫模型研究进展.中华神经外科杂志, 2014, 30(1):88-90.
15 陈树达,陈子怡,方子妍,等.普朗尼克P85与维拉帕米对苯妥英钠脑靶向分布的比较.中山大学学报(医学科学版), 2014, 35(2):161-168.