柴贝止痫汤及其主要入血成分柴胡皂苷a对癫痫模型大鼠卡马西平脑内分布的影响
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摘要
目的观察柴贝止痫汤及柴胡皂苷a治疗难治性癫痫的可能作用机制。方法 88只大鼠随机分为空白组12只及模型组、卡马西平组、柴贝止痫汤+卡马西平组、柴胡皂苷a+卡马西平组19只。除空白组外,其余各组采用侧脑室注射海人酸制备癫痫大鼠模型。卡马西平组给予卡马西平混悬液0.75 ml/(100 g·d),柴贝止痫汤+卡马西平组给予柴贝止痫汤0.5 ml/(100 g·d)和卡马西平混悬液0.75 ml/(100 g·d),柴胡皂苷a+卡马西平组给予柴胡皂苷a混悬液0.25 ml/100 g和卡马西平混悬液0.75 ml/100 g,模型组及空白组给予等量羧甲基纤维素钠混悬液。给药60天后比较各组海马P糖蛋白、Mdr1a mRNA表达及全脑卡马西平(CBZ)、1011-环氧化卡马西平(CBZE)含量。结果与空白组比较,模型组海马P糖蛋白表达升高(P<0.05);与模型组比较,卡马西平组、柴胡皂苷a+卡马西平组海马P糖蛋白表达差异无统计学意义(P>0.05),柴贝止痫汤+卡马西平组海马P糖蛋白表达降低(P<0.05);与卡马西平组比较,柴贝止痫汤+卡马西平组和柴胡皂苷a+卡马西平组海马P糖蛋白表达均降低(P<0.05),柴贝止痫汤+卡马西平组和柴胡皂苷a+卡马西平组海马P糖蛋白表达差异无统计学意义(P>0.05)。模型组Mdr1a mRNA表达较空白组升高。卡马西平组的Mdr1a mRNA表达高于空白组(P<0.05),空白组、模型组MDR1a mRNA表达分别为卡马西平组的0.36倍、0.87倍。柴贝止痫汤+卡马西平组较卡马西平组Mdr1a含量降低(P<0.01),为卡马西平组的0.14倍。与卡马西平组比较,柴贝止痫汤+卡马西平组CBZE含量升高(P<0.01),柴胡皂苷a+卡马西平组CBZE含量降低(P<0.01),且柴贝止痫汤+卡马西平组和柴胡皂苷a+卡马西平组CBZ差异无统计学意义(P>0.05);与柴贝止痫汤+卡马西平组比较,柴胡皂苷a+卡马西平组CBZ、CBZE含量均降低(P<0.01)。结论柴贝止痫汤联合卡马西平可降低癫痫模型大鼠海马Mdr1a/P糖蛋白表达,促进CBZ、CBZE入脑,对CBZE作用突出,柴胡皂苷a在其中不发挥主要作用。
Objective To observe the possible mechanism of Chaibei Zhixian Decoction( CBZXD,柴贝止痫汤)and Saikosaponin a( SSa) in treating intractable epilepsy. Methods A total of 88 rats were randomly divided into blank group of 12 rats and every 19 rats for model group,carbamazepine( CBZE) group,the CBZXD + CBZE group,and the saikosaponin a + CBZE group. Except for the blank group,other groups used the lateral ventricle injection of kainate to prepare a rat model of epilepsy. The CBZE group was given CBZE suspension 0. 75 ml/( 100 g·d). The CBZXD + CBZE group was given CBZXD 0. 5 ml/( 100 g·d) and CBZE suspension 0. 75 ml/( 100 g·d).The saikosaponin a + CBZE group was given saikosaponin a suspension 0. 25 ml/100 g and CBZE suspension0. 75 ml/100 g. The model group and the blank group were given the same amount sodium carboxymethylcellulose suspension. After 60 days of administration,the expression of P-glycoprotein,Mdr1 a mRNA and the content of whole brain carbamazepine( CBZ) and 10 11-epoxidized CBZE were compared. Results Compared with the blank group,the expression of P-glycoprotein in the hippocampus of the model group was increased( P < 0. 05). Compared with the model group,the expression of P-glycoprotein in the hippocampus of the CBZE group,saikosaponin a + carbazepine group was not statistically significant( P > 0. 05). The expression of P-glycoprotein in hippocampus of CBZXD + CBZE group was decreased( P < 0. 05). Compared with CBZE group,the expression of P-glycoprotein in hippocampus of CBZXD + CBZE group and saikosaponin a + CBZE group was decreased( P < 0. 05). There was no significant difference in the expression of P-glycoprotein in hippocampus between CBZXD + CBZE group and saikosaponin a + CBZE group( P > 0. 05). Mdr1 a mRNA expression in the model group was increased compared with that in the blank group. The expression of Mdr1 a mRNA in CBZE group was higher than that in the blank group( P <0. 05). The expression of MDR1 a mRNA in blank group and model group was 0. 36 times and 0. 87 times higher than that in CBZE group. The content of Mdr1 a in the CBZXD + CBZE group was decreased when compared with that in the CBZE group( P < 0. 01),which was 0. 14 times higher than that in the CBZE group. Compared with CBZE group,CBZE content in CBZXD + CBZE group was increased( P < 0. 01). CBZE content in saikosaponin a +CBZE group was decreased( P < 0. 01),and there was no significant difference in CBZ between CBZXD + CBZE group and saikosaponin a + CBZE group( P > 0. 05). Compared with CBZXD + CBZE group,saikosaponin a +CBZE group,the content of CBZ,CBZE was reduced( P < 0. 01). Conclusion CBZXD combined with CBZE can reduce the expression of Mdr1 a/P glycoprotein in hippocampus of rats with epilepsy,promote CBZ and CBZE into the brain,and play an important role in CBZE. The saikosaponin a dose not play a major role in the above process.
Objective To observe the possible mechanism of Chaibei Zhixian Decoction( CBZXD,柴贝止痫汤)and Saikosaponin a( SSa) in treating intractable epilepsy. Methods A total of 88 rats were randomly divided into blank group of 12 rats and every 19 rats for model group,carbamazepine( CBZE) group,the CBZXD + CBZE group,and the saikosaponin a + CBZE group. Except for the blank group,other groups used the lateral ventricle injection of kainate to prepare a rat model of epilepsy. The CBZE group was given CBZE suspension 0. 75 ml/( 100 g·d). The CBZXD + CBZE group was given CBZXD 0. 5 ml/( 100 g·d) and CBZE suspension 0. 75 ml/( 100 g·d).The saikosaponin a + CBZE group was given saikosaponin a suspension 0. 25 ml/100 g and CBZE suspension0. 75 ml/100 g. The model group and the blank group were given the same amount sodium carboxymethylcellulose suspension. After 60 days of administration,the expression of P-glycoprotein,Mdr1 a mRNA and the content of whole brain carbamazepine( CBZ) and 10 11-epoxidized CBZE were compared. Results Compared with the blank group,the expression of P-glycoprotein in the hippocampus of the model group was increased( P < 0. 05). Compared with the model group,the expression of P-glycoprotein in the hippocampus of the CBZE group,saikosaponin a + carbazepine group was not statistically significant( P > 0. 05). The expression of P-glycoprotein in hippocampus of CBZXD + CBZE group was decreased( P < 0. 05). Compared with CBZE group,the expression of P-glycoprotein in hippocampus of CBZXD + CBZE group and saikosaponin a + CBZE group was decreased( P < 0. 05). There was no significant difference in the expression of P-glycoprotein in hippocampus between CBZXD + CBZE group and saikosaponin a + CBZE group( P > 0. 05). Mdr1 a mRNA expression in the model group was increased compared with that in the blank group. The expression of Mdr1 a mRNA in CBZE group was higher than that in the blank group( P <0. 05). The expression of MDR1 a mRNA in blank group and model group was 0. 36 times and 0. 87 times higher than that in CBZE group. The content of Mdr1 a in the CBZXD + CBZE group was decreased when compared with that in the CBZE group( P < 0. 01),which was 0. 14 times higher than that in the CBZE group. Compared with CBZE group,CBZE content in CBZXD + CBZE group was increased( P < 0. 01). CBZE content in saikosaponin a +CBZE group was decreased( P < 0. 01),and there was no significant difference in CBZ between CBZXD + CBZE group and saikosaponin a + CBZE group( P > 0. 05). Compared with CBZXD + CBZE group,saikosaponin a +CBZE group,the content of CBZ,CBZE was reduced( P < 0. 01). Conclusion CBZXD combined with CBZE can reduce the expression of Mdr1 a/P glycoprotein in hippocampus of rats with epilepsy,promote CBZ and CBZE into the brain,and play an important role in CBZE. The saikosaponin a dose not play a major role in the above process.
引文
[1]GAO L,XIA L,ZHAO FL,et al.Clinical efficacy and safety of the newer antiepileptic drugs as adjunctive treatment in adults with refractory partial-onset epilepsy:a meta-analysis of randomized placebo-controlled trials[J].Epilepsy Res,2013,103(1):31-44.
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[3]郑香春,李淑芳,刘金民.柴贝止痫汤单药及合并用药治疗难治性癫痫临床观察[J].天津中医药,2012,29(3):224-227.
[4]聂莉媛,鄢泽然,张青,等.柴贝止痫汤添加治疗难治性癫痫复杂部分性发作的临床研究[J].环球中医药,2015,8(1):13-18.
[5]KANDRATAVICIUS L,BALISTA PA,LOPES-AGUIAR C,et al.Animal models of epilepsy:use and limitations[J].Neuropsychiatr Dis Treat,2014,10:1693-1705.doi:10.2147/NDT.S50371.
[6]PAXINOS G,WATSON C.The Rat Brain:In Stereotaxic Coordinates[M].New York:Academic Press,1982:1-104.
[7]RACINE JR.Modification of seizure activity by electrical stimulation:ⅡMotor seizures[J].Electroencephalogr Clin Neurophysiol,1972,32(3):269-279.
[8]崔燎.药理学实验教程[M].北京:科学出版社,2011:8.
[9]ENGEL J JR.Mesial temporal lobe epilepsy:what have we learned?[J].Neuroscientist,2001,7(4):340-352.
[10]VARMA MV,ASHOKRAI Y,DEY CS,et al.P-glycoprotein inhibitors and their screening:a perspective from bioavailability enhancement[J].Pharmacol Res,2003,48(4):347-359.
[11]FISCHER V,EINOLF HJ,COHEN D.Efflux transporters and their clinical relevance[J].Mini Rev Med Chem,2005,5(2):183-195.
[12]NOBILI S,LANDINI I,MAZZEI T,et al.Overcoming tumor multidrug resistance using drugsable to evade P-glycoprotein or to exploit its expression[J].Med Res Rev,2012,32(6):1220-1262.
[13]DOMBROWSKI SM,DESAI SY,MARRONI M,et al.Overexpression of multiple drug resistancegenes in endothelial cells from patients with refractory epilepsy[J].Epilepsia,2010,42(12):1501-1506.
[14]BARTMANN H,FUEST C,LA FOUGERE C,et al.Imaging of P-glycoprotein-mediated pharmacoresistance in the hippocampus:proof-of-concept in a chronic rat model of temporal lobe epilepsy[J].Epilepsia,2010,51(9):1780-1790.
[15]VOLK HA,LOSCHER W.Multidrug resistance in epilepsy:rats with drug-resistant seizures exhibit enhanced brain expression of P-glycoprotein compared with rats with drug-responsive seizures[J].Brain,2005,128(6):1358-1368.
[16]肖争,晏勇.大鼠耐苯妥英钠和苯巴比妥钠杏仁核点燃模型及多药耐受基因的表达[J].中华神经科杂志,1999,32(6):365-368.
[17]WEN T,LIU YC,YANG HW,et al.Effect of 21-day exposure of phenobarbital,carbamazepine and phenytoin on P-glycoprotein expression and activity in the rat brain[J].J Neurol Sci,2008,270(1):99-106.
[18]WANG Y,ZHOU D,WANG B,et al.A kindling model of pharmacoresistant temporal lobe epilepsy in spraguedawley rats induced by coriaria lactone and its possible mechanism[J].Epilepsia,2010,44(4):475-488.
[19]李淑芳.颞叶癫痫中医证候研究及柴贝止痫汤对癫痫大鼠GABAR、NMDAR1表达的影响[D].北京:北京中医药大学,2009:58.
[20]王越,刘金民.中药柴贝止痫汤对难治性癫痫大鼠模型行为学及P-糖蛋白和MDR1表达的影响[J].中华中医药杂志,2017,32(4):1735-1738.
[21]牛倩倩,陈梦钰,曹博群,等.脑心清对卡马西平在癫痫大鼠体内药动学和脑组织分布的影响[J].中药药理与临床,2018,34(1):114-117.
[22]卜书红,张健,陆晓彤,等.儿童癫痫患者CYP酶活性与血清卡马西平浓度和临床疗效的相关性分析[J].医药导报,2010,29(4):430-432.
[23]CHEN SY,WANG K,Wan YJY.Retinoids activate RXR/CAR-mediated pathway and induce CYP3A[J].Biochem Pharmacol,2010,79(2):270-276.
[24]王永辉.柴胡总皂苷对小鼠部分药物代谢酶及P-糖蛋白的影响[D].广州:广州中医药大学,2012:33-35.
[25]谢炜,林佳,张作文,等.柴胡皂苷a对体外培养大鼠海马星形胶质细胞激活的抑制作用[J].南方医科大学学报,2008,28(10):1798-1801.
[26]YU YH,WEI X,YONG B,et al.Saikosaponin a mediates the anticonvulsant properties in the HNC models of AE and SE by inhibiting NMDA receptor current and persistent sodium current[J].Plo S One,2012,7(11):e50694.
[27]YE M,BI YF,DING L,et al.Saikosaponin a functions as anti-epileptic effect in pentylenetetrazol induced rats through inhibiting m TOR signaling pathway[J].Biomed Pharmacother,2016,81:281-287.doi:10.1016/j.biopha.2016.04.012.
[2]ZHANG C,KWAN P,ZUO Z,et al.The transport of antiepileptic drugs by P-glycoprotein[J].Adv Drug Deliv Rev,2012,64(10):930-942.
[3]郑香春,李淑芳,刘金民.柴贝止痫汤单药及合并用药治疗难治性癫痫临床观察[J].天津中医药,2012,29(3):224-227.
[4]聂莉媛,鄢泽然,张青,等.柴贝止痫汤添加治疗难治性癫痫复杂部分性发作的临床研究[J].环球中医药,2015,8(1):13-18.
[5]KANDRATAVICIUS L,BALISTA PA,LOPES-AGUIAR C,et al.Animal models of epilepsy:use and limitations[J].Neuropsychiatr Dis Treat,2014,10:1693-1705.doi:10.2147/NDT.S50371.
[6]PAXINOS G,WATSON C.The Rat Brain:In Stereotaxic Coordinates[M].New York:Academic Press,1982:1-104.
[7]RACINE JR.Modification of seizure activity by electrical stimulation:ⅡMotor seizures[J].Electroencephalogr Clin Neurophysiol,1972,32(3):269-279.
[8]崔燎.药理学实验教程[M].北京:科学出版社,2011:8.
[9]ENGEL J JR.Mesial temporal lobe epilepsy:what have we learned?[J].Neuroscientist,2001,7(4):340-352.
[10]VARMA MV,ASHOKRAI Y,DEY CS,et al.P-glycoprotein inhibitors and their screening:a perspective from bioavailability enhancement[J].Pharmacol Res,2003,48(4):347-359.
[11]FISCHER V,EINOLF HJ,COHEN D.Efflux transporters and their clinical relevance[J].Mini Rev Med Chem,2005,5(2):183-195.
[12]NOBILI S,LANDINI I,MAZZEI T,et al.Overcoming tumor multidrug resistance using drugsable to evade P-glycoprotein or to exploit its expression[J].Med Res Rev,2012,32(6):1220-1262.
[13]DOMBROWSKI SM,DESAI SY,MARRONI M,et al.Overexpression of multiple drug resistancegenes in endothelial cells from patients with refractory epilepsy[J].Epilepsia,2010,42(12):1501-1506.
[14]BARTMANN H,FUEST C,LA FOUGERE C,et al.Imaging of P-glycoprotein-mediated pharmacoresistance in the hippocampus:proof-of-concept in a chronic rat model of temporal lobe epilepsy[J].Epilepsia,2010,51(9):1780-1790.
[15]VOLK HA,LOSCHER W.Multidrug resistance in epilepsy:rats with drug-resistant seizures exhibit enhanced brain expression of P-glycoprotein compared with rats with drug-responsive seizures[J].Brain,2005,128(6):1358-1368.
[16]肖争,晏勇.大鼠耐苯妥英钠和苯巴比妥钠杏仁核点燃模型及多药耐受基因的表达[J].中华神经科杂志,1999,32(6):365-368.
[17]WEN T,LIU YC,YANG HW,et al.Effect of 21-day exposure of phenobarbital,carbamazepine and phenytoin on P-glycoprotein expression and activity in the rat brain[J].J Neurol Sci,2008,270(1):99-106.
[18]WANG Y,ZHOU D,WANG B,et al.A kindling model of pharmacoresistant temporal lobe epilepsy in spraguedawley rats induced by coriaria lactone and its possible mechanism[J].Epilepsia,2010,44(4):475-488.
[19]李淑芳.颞叶癫痫中医证候研究及柴贝止痫汤对癫痫大鼠GABAR、NMDAR1表达的影响[D].北京:北京中医药大学,2009:58.
[20]王越,刘金民.中药柴贝止痫汤对难治性癫痫大鼠模型行为学及P-糖蛋白和MDR1表达的影响[J].中华中医药杂志,2017,32(4):1735-1738.
[21]牛倩倩,陈梦钰,曹博群,等.脑心清对卡马西平在癫痫大鼠体内药动学和脑组织分布的影响[J].中药药理与临床,2018,34(1):114-117.
[22]卜书红,张健,陆晓彤,等.儿童癫痫患者CYP酶活性与血清卡马西平浓度和临床疗效的相关性分析[J].医药导报,2010,29(4):430-432.
[23]CHEN SY,WANG K,Wan YJY.Retinoids activate RXR/CAR-mediated pathway and induce CYP3A[J].Biochem Pharmacol,2010,79(2):270-276.
[24]王永辉.柴胡总皂苷对小鼠部分药物代谢酶及P-糖蛋白的影响[D].广州:广州中医药大学,2012:33-35.
[25]谢炜,林佳,张作文,等.柴胡皂苷a对体外培养大鼠海马星形胶质细胞激活的抑制作用[J].南方医科大学学报,2008,28(10):1798-1801.
[26]YU YH,WEI X,YONG B,et al.Saikosaponin a mediates the anticonvulsant properties in the HNC models of AE and SE by inhibiting NMDA receptor current and persistent sodium current[J].Plo S One,2012,7(11):e50694.
[27]YE M,BI YF,DING L,et al.Saikosaponin a functions as anti-epileptic effect in pentylenetetrazol induced rats through inhibiting m TOR signaling pathway[J].Biomed Pharmacother,2016,81:281-287.doi:10.1016/j.biopha.2016.04.012.
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