穴位埋药线对难治性癫痫大鼠癫痫样波的发放及大脑海马和颞叶皮质多药耐药相关蛋白MRP_1、P-gp表达的影响

Effects of Acupoint Embedding Therapy on Epileptiform Wave of Intractable Epilepsy Rats and the MRP_1, P-gp Protein in Hippocampus and Temporal Cortex

目的探讨穴位埋药线对难治性癫痫大鼠癫痫样波的发放及大脑海马和皮质中多药耐药相关蛋白MRP1、P-gp表达的影响。方法 (1)造模:大鼠海马区注射红藻氨酸(KA),经过点燃和再次亚惊厥剂量点燃造模,且脑电图检测有癫痫样波发放者筛选为造模成功耐药难治性癫痫模型鼠。(2)分组与处理:普通线组(PTX组)与药线组(YX组),先埋一侧穴位,间隔15 d后埋对侧穴位。拉莫三嗪组(LTG组)按照人用拉莫三嗪剂量换算灌胃大鼠,每日2次;空白对照组(Normal组)和模型组(Model组)给予灌胃同等体积的蒸馏水,均灌胃30 d。(3)采用VEEG-1518K型数字化视频脑电监测分析系统检测脑波基本节律的波幅与频率变化。(4)标本处理与检测:采用免疫组织化学技术,检测多药耐药相关蛋白MRP1、P-gp在海马与颞叶皮层不同部位的表达。结果各组治疗前比较差异无统计学意义(P>0.05);各组治疗后,YX组、LTG组分别与Model组、PTX组比较差异有统计学意义(P<0.05),YX组与LTG组比较差异无统计学意义(P>0.05);与Model组相比,LTG组和YX组干预后能降低致痫鼠EEG痫波放电持续时间与发放频率以及海马区和颞叶皮质区多药耐药蛋白MRP1、P-gp的表达水平(P<0.05或P<0.01);YX组与PTG组比较差异无统计学意义(P<0.05)。结论(1)埋药线使KA致痫鼠EEG痫波放电持续时间缩短,发放频率减少。(2)KA点燃难治性癫痫模型大鼠大脑海马区存在多药耐药蛋白MRP1、P-gp的表达较皮质区明显升高。(3)埋药线逆转与降低致痫鼠海马区多药耐药蛋白MRP1、P-gp的表达水平,可能是其抗癫痫生物学作用机制之一。

Objective To discussion the acupoint embedding therapy on epileptiform wave of intractable epilepsy rats and the multidrug resistance protein 1（MRP1）、 P-gp protein in hippocampus and temporal cortex. Methods（1） Making models：The rats were injected with kainic acid（KA） in hippocampus region, which kinded and rekinded of sub-convulsant doseto making model. The rats with EEG epileptiform wave are screened as successful modeling refractory epilepsy rats.（2）Grouping and treatment： Buried ordinary wire line group（PTX group） and the drug line group（YX group） were firstly buried side points, spaced 15 days buried in the contralateral acupoints. Lamotrigine（LTG） group rats were fed 2 times a day.The model group were administered the same volume gavage of distilled water for 30 days.（3） The changes of EEG basic rhythm amplitude and frequency were detectd by using VEEG-1518 K type digital video EEG monitoring and analysis system.（4） Selecting and detecting specimens： The expressions of multi-resistant related protein MRP1 and P-gp in the hippocampus and temporal lobe cortex were detected. Results The expressions of each group before treatment had no significant difference（P〈0.05）. After treatment, The YX group and LTG group were compared with Model group and PTX group, respectively,the difference was statistically significant（P〈0.05）, while YX group and LTG group had no significant difference（P〈0.05）.Compared with model group, LTG group and YX group can reduce epileptiform EEG wave discharge duration and frequency of issuance in epileptic rats and the multidrug resistance protein MRP1, P-gp expression level in hippocampus and temporal cortex（P〈0.05 or P〈0.01）, the difference between the YX group and PTX group was statistically significant（P〈0.05）.Conclusion（1） Embedding medicine can short the EEG wave discharge duration time and reduce the frequency of epileptic rats induced by KA seizures.（2） The expression of multidrug resistance related prontein MRP1 and P-gp in hippocampus were obviously higher than that in cortical areas of KA induced epileptic rats.（3） The reversal and reduction of the expression levels of multidrug resistance gene hippocampus of epileptic rats may be one of the mechanisms of embedding medicine on anti-epilepsy.