疏风止痉方对耐药癫痫大鼠发作行为及NF-κB通路影响

Effect of Shufeng Zhijing Decoction on Seizure Behavior and NF-kappa B Pathway in Drug-resistant Epilepsy Rats

目的:基于NF-κB通路探讨疏风止痉方对耐药性癫痫大鼠的发作机制。方法:健康幼龄SD大鼠100只,随机选取10只作为正常组大鼠,其余90只用于匹罗卡品点燃癫痫模型,将点燃成功的癫痫模型大鼠通过2周卡马西平、2周丙戊酸钠预处理后仍有癫痫发作的大鼠作为耐药性癫痫模型。将耐药性癫痫模型大鼠随机分5组,分别予卡马西平(Carbamazepine,CBZ)、特异性抑制剂(Pyrrolidine dithiocarbamate,PDTC)、卡马西平联合疏风止痉方低剂量、卡马西平联合疏风止痉方中剂量、卡马西平联合疏风止痉方高剂量灌胃治疗28 d,观察大鼠癫痫发作情况,免疫组化法检测及免疫印记蛋白(Western blot)法检测核转录因子κB(Nuclear factor kappa-B,NF-κB)、P糖蛋白(P-glycoprotein,P-gp)及NF-κB信号通路上下游细胞因子的表达。结果:正常组大鼠未见发作,海马NF-κB、P-gp及相关炎性因子表达最低。与正常组比较,CBZ组、PDTC组、疏卡联合组发作次数减少(P<0.01),各实验组NF-κB、P-gp及相关炎性因子表达增强(P<0.05)。与CBZ组比较,PDTC组、疏卡高组发作次数明显减少(P<0.01),疏卡高组发作级别降低(P<0.05),发作时间无差异(P>0.05),NF-κB、P-gp及相关炎性因子表达降低(P<0.01),PDTC组在降低NF-κB方面更有优势。与PDTC组比较,疏卡高组发作次数减少(P<0.05),发作级别降低(P<0.05),NF-κB、P-gp表达增强(P<0.01),相关炎性因子表达降低(P<0.05),CBZ组发作次数增多、发作级别升高(P<0.05),NF-κB、P-gp表达增强(P<0.01),相关炎性因子表达无差异(P>0.05)。结论:疏风止痉方可以减少耐药性癫痫模型大鼠发作次数、减轻发作级别,对发作持续时间无影响,其机制可能与疏风止痉方抑制相关炎性因子的表达,下调NF-κB、P-gp水平有关。

Objective:To explore the mechanism of Shufeng Zhijing decoction on drug-resistant epilepsy rats based on the NF-kappa B pathway.Methods:100 healthy young SD rats were randomly selected as normal group,and the remaining 90 rats were used as pilocarpine kindling epilepsy model.The successful kindling epilepsy model rats were used as drug-resistant epilepsy model after 2 weeks of pretreatment with carbamazepine and 2 weeks of sodium valproate.Drug-resistant epilepsy model rats were randomly divided into five groups.They were given Carbamazepine(CBZ),Pyrrolidine dithiocarbamate(PDTC),Carbamazepine combined with Shufeng Zhicong decoction at low dose,Carbamazepine combined with Shufeng Zhicong decoction at medium dose and Carbamazepine combined with Shufeng Zhicong decoction at high dose for 28 days respectively.The seizures and immunity of rats were observed.The expression of downstream cytokines in nuclear factor kappa-B(NF-kappa-B),P-glycoprotein(P-gp)and NFkappa B signaling pathway was detected by histochemical method and Western blot.Results:There was no seizure in the normal group,and the expression of NF-kappa B,P-gp and related inflammatory factors in hippocampus was the lowest.Compared with the normal group,the number of seizures in CBZ group,PDTC group and Shuka combined group decreased(P<0.01),and the expression of NF-kappa B,P-gp and related inflammatory factors increased(P<0.05).Compared with CBZ group,the number of attacks in PDTC group and Shukagao group decreased significantly(P<0.01),the level of attacks in Shukagao group decreased(P<0.05),the duration of attacks had no difference(P>0.05),and the expression of NF-kappa B,P-gp and related inflammatory factors decreased(P<0.01).PDTC group had more advantages in reducing NF-kappa B.Compared with the PDTC group,the frequency of seizures decreased(P<0.05),the level of seizures decreased(P<0.05),the expression of NF-kappa B and P-gp increased(P<0.01),the expression of related inflammatory factors decreased(P<0.05),the number of seizures increased,the level of seizures increased(P<0.05),the expression of NF-kappa B and P-gp increased(P<0.01),and the expression of related inflammatory factors had no difference(P>0.05).Conclusion:Shufeng Zhiquan Decoction can reduce the number of seizures and the level of seizures in drug-resistant epilepsy model rats,and has no effect on the duration of seizures.The mechanism may be related to the inhibition of the expression of inflammatory factors and the down-regulation of the levels of NF-kappa B and P-gp by Shufeng Zhiquan Decoction.