柴贝止痫汤及其吸收成分贝母素甲对大鼠脑内卡马西平含量的影响及其机制

Effect of Chaibei Zhixian Decoction and Its Absorption Component Peimine on Content of Carbamazepine in Rat Brain and Investigation of Its Mechanism

目的:观察柴贝止痫汤及贝母素甲(Peimine)对难治性癫痫大鼠脑内卡马西平(CBZ)含量及P-糖蛋白(P-gp)和多药耐药基因1(MDR1)表达的影响,了解贝母素甲在该复方中抗难治性癫痫的贡献度。方法:该研究采用侧脑室注射海人酸的方法制备难治性癫痫大鼠模型,造模成功的大鼠随机分为模型组,CBZ组(0. 12 g·kg-1),柴贝止痫汤+CBZ组(8. 39 g·kg-1+0. 12 g·kg-1),贝母素甲+CBZ组(0. 01 g·kg-1+0. 12 g·kg-1),并设假手术组,治疗60 d后,采用蛋白免疫印迹法(Western blot),实时荧光定量聚合酶链式反应(Real-time PCR),液相色谱-质谱联用(LC-MS)方法检测大鼠脑内皮层中P-gp/MDR1的表达,及脑内CBZ,10,11-环氧化卡马西平(CBZE)含量的变化。结果:与假手术组比较,模型组大鼠皮层中P-gp/MDR1表达明显升高(P <0. 05,P <0. 01);与模型组比较,CBZ组大鼠皮层中P-gp/MDR1表达升高;柴贝止痫汤+CBZ组和贝母素甲+CBZ组大鼠皮层中P-gp/MDR1表达降低;与CBZ组比较,柴贝止痫汤+CBZ组和贝母素甲+CBZ组大鼠皮层中P-gp/MDR1表达明显降低(P <0. 05,P <0. 01),贝母素甲+CBZ组大鼠脑内CBZ和CBZE含量升高(P <0. 05,P <0. 01),柴贝止痫汤+CBZ组大鼠脑内CBZE含量明显升高(P <0. 05);与柴贝止痫汤+CBZ组比较,贝母素甲+CBZ组大鼠脑内CBZ和CBZE含量升高,差异没有统计学意义。结论:柴贝止痫汤及贝母素甲可能通过降低难治性癫痫大鼠皮层P-gp/MDR1表达,增加脑内CBZ和CBZE含量。

Objective:To observe the effect of Chaibei Zhixian decoction and peimine on Carbamazepine(CBZ)concentration,P-glycoprotein(P-gp)and multi drug resistance 1(MDR1)expression in the brain tissues of rats with refractory epilepsy,and to understand the contribution of Peimine in the compound prescription to treat the refractory epilepsy.Method:Epilepsy rat models were established by injecting kainic acid(KA)in the lateral ventricle.The successfully modeled rats were randomly divided into model group,CBZ group(0.12 g·kg-1),Chaibei Zhixian decoction+CBZ group(8.39 g·kg-1+0.12 g·kg-1),peimine+CBZ group(0.01 g·kg-1+0.12 g·kg-1)and sham operation group.After 60 days of intervention,the expression levels of P-glycoprotein(Pgp)and MDR1 b mRNA in the brain cortex were detected by Western blot and quantitative real-time fluorescence polymerase chain reaction(Real-time PCR),the contents of CBZ and 10,11-epoxidation of carbamazepine(CBZE)were measured by liquid chromatography-mass spectrometry(LC-MS).Result:Compared with sham group,the expression of P-gp/MDR1 in the cortex of model group was significantly increased(P<0.05,P<0.01).Compared with model group,the P-gp/MDR1 level in CBZ group was increased.The expression of P-gp/MDR1 in the cortex of Chaibei Zhixian decoction+CBZ group and peimine+CBZ group was reduced.Compared with CBZ group,The expression of P-gp/MDR1 was significantly decreased in the cortex of Chaibei Zhixian decoction+CBZ group and peimine+CBZ group(P<0.05,P<0.01),and the content of CBZ and CBZE in the brain of peimine+CBZ group was significantly increased(P<0.05,P<0.01),and the content of CBZE in the brain of the Chaibei Zhixian decoction+CBZ group was significantly increased(P<0.05).Compared with the Chaibei Zhixian decoction+CBZ group,the content of CBZ and CBZE in the brain of the peimine+CBZ group was increased in rats,but the difference was not statistically significant.Conclusion:Chaibei Zhixian decoction and peimine may increase the content of CBZ and CBZE in the brain tissues in rats with intractable epilepsy by reducing the expression of MDR1/P-gp in the cortex.