柴贝止痫汤对难治性癫痫大鼠多药耐药蛋白P-gp表达影响的研究
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摘要
目的通过比较不同药物干预对多药耐药蛋白P-gp的影响,观察难治性癫痫大鼠脑内P-gp的表达和分布,探讨难治性癫痫多药耐药可能的分子病理机制,及中药复方柴贝止痫汤对其表达的影响。方法 80只Wistar大鼠腹腔注射氯化锂-盐酸匹罗卡品建立难治性癫痫模型,12只同周龄Wistar大鼠作为正常组。筛选出59只造模成功大鼠,随机分为模型组14只,柴贝止痫汤组(15只;8. 1 g/kg灌胃),卡马西平组(15只; 36 mg/kg灌胃)和联合用药组(15只;柴贝止痫汤和卡马西平联合灌胃)。干预2周后观察各致痫组大鼠癫痫发作的平均持续时间、癫痫发作级别,以判定、分析药物的疗效。采用免疫组织化学和蛋白质印迹法,分析、比较P-gp在各组大鼠海马和皮质等部位的表达和分布。结果与正常组大鼠比较,在难治性癫痫大鼠海马和皮质部位,各致痫组P-gp表达含量均有明显增高,具有统计学意义(P <0. 05)。结论柴贝止痫汤联合卡马西平的给药方式对难治性癫痫大鼠的疗效优于单独应用卡马西平及柴贝止痫汤。中药复方柴贝止痫汤具有一定的抑制P-gp药物转运体功能。
Objective By investigating the effects of Chaibei Zhixian( Radix Bupleuri and Bulbus Fritillariae Thunbergii anti-epilepsy,CBZX) decoction,a Chinese herbal compound,on the expression and distribution of multidrug resistance protein( P-glycoprotein,P-gp) in intractable epileptic rats,to explore the possible molecular pathogenesis mechanism of intractable epilepsy. Methods 80 Wistar rats were intraperitoneally injected lithium chloride-pilocarpine hydrochloride to establish intractable epilepsy model,and 12 rats at the same week age were used as the normal control. 59 rats were selected as successful model and randomly divided into model group( n = 14),CBZX group( n = 15; 8. 1 g/kg intragastric gavage),and carbamazepine( CBZ) group( n = 15; 36 mg/kg intragastric gavage) and combination treatment group( n =15; CBZX plus CBZ intragastric gavage). The average duration of seizures and the Racine's grade of epileptic seizure were evaluated after 2 weeks' intervention. Immunohistochemical method and Western blotting assay were used to compare the expression and distribution of P-gp in the hippocampus and cerebral cortex of the rats. Results Compared with the normal group,the expression of P-gp increased significantly in all epileptic groups( P < 0. 05). Conclusion CBZX decoction combined with carbamazepine is superior to using single therapy. The Chinese herbal compound CBZX decoction seems to show some effects in inhibiting P-gp drug transporters.
Objective By investigating the effects of Chaibei Zhixian( Radix Bupleuri and Bulbus Fritillariae Thunbergii anti-epilepsy,CBZX) decoction,a Chinese herbal compound,on the expression and distribution of multidrug resistance protein( P-glycoprotein,P-gp) in intractable epileptic rats,to explore the possible molecular pathogenesis mechanism of intractable epilepsy. Methods 80 Wistar rats were intraperitoneally injected lithium chloride-pilocarpine hydrochloride to establish intractable epilepsy model,and 12 rats at the same week age were used as the normal control. 59 rats were selected as successful model and randomly divided into model group( n = 14),CBZX group( n = 15; 8. 1 g/kg intragastric gavage),and carbamazepine( CBZ) group( n = 15; 36 mg/kg intragastric gavage) and combination treatment group( n =15; CBZX plus CBZ intragastric gavage). The average duration of seizures and the Racine's grade of epileptic seizure were evaluated after 2 weeks' intervention. Immunohistochemical method and Western blotting assay were used to compare the expression and distribution of P-gp in the hippocampus and cerebral cortex of the rats. Results Compared with the normal group,the expression of P-gp increased significantly in all epileptic groups( P < 0. 05). Conclusion CBZX decoction combined with carbamazepine is superior to using single therapy. The Chinese herbal compound CBZX decoction seems to show some effects in inhibiting P-gp drug transporters.
引文
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[2]Kwan P,Arzimanoglou A,Berg A T,et al. Definition of drug resistant epilepsy:Consensus proposal by the ad hoc Task Force of the ILAE Commission on Therapeutic Strategies[J]. Epilepsia,2010,51(6):1069-1077.
[3] Dewanjee S,Dua TK,Bhattacharjee N,et al. Natural Products as Alternative Choices for P-Glycoprotein(P-gp)Inhibition[J]. Molecules,2017,22(6):E871.
[4] Racine RJ. Modification of seizure activity by electrical stimulation:Ⅱ. Motor seizures[J]. Electroencephalogr.Clin Neurophysiol,1972,32(3):269-279.
[5]陈鸿雁,王弛,舒燕,等.抗肿瘤药物与维拉帕米联合对KBV-200细胞P-糖蛋白表达影响的研究[J].重庆医科大学学报,2006,31(3):369-372.Chen HY,Wang C,Shu Y,et al. Investigation of the effect of verapamil and chemotherapeutic agents on the Pglucoprotein of the KBV200 cell line[J]. Journal of Chongqing Medical University,2006,31(3):369-372.
[6]Shitan N,Tanaka M,Tepai K,et al. Human MDR1 and MRP1 recognize berberine as their transport substrate[J].Biosci Biotechnol Biochem,2007,71(1):242-245.
[7]Raad I,Terreux R,Richomme P,et al. Structure activity relationship of natural and synthetic coumarins inhibition the multidrug transporter P-glycoprotein[J]. Bioorg Med Chem,2006,14(20):6979-6987.