基于NF−κB炎症信号通路探讨疏风止痉方逆转耐药性癫痫大鼠的机制

Drug-resistant epilepsy is an intractable problem in the field of epilepsy treatment. In the preliminary clinical study, we found that adding dispersing wind and relieving convulsion formula like Yinqiao Powder was effective for drug-resistant epilepsy. According to that Yinqiao Powder could regulate inflammatory reaction, and that recent researches have confirmed the relation line between "repeated seizures, activate NF -κB inflammatory signaling pathways, up-regulated expression of P-gp, and drug-resistant epilepsy”, we put forward the hypothesis that: “ the potential mechanism of dispersing wind and relieving convulsion formula reversing treating drug-resistant epilepsy is: regulating NF-κB inflammatory signaling pathways, and thus inhibit the up-regulation of P-gp, which improves the concentration of anti-epilepsy drugs in brain, reverses the drug-resistant epilepsy, and thus improve the clinical efficacy”. This study intends to base on the breakthrough points of “inflammatory reaction in epilepsy” and main line of epileptic drug-resistant mechanism. In this study,after drug interventin on all group ,we will make drug resistant rat models, detect and analyze relative indexes like after discharge threshold value, rat onste, drug concentration in the cerebrospinal fluid, NF-κB protein, relative cytokines, P-gp in brain district, to prove the therapeutic effect of adding dispersing wind and relieving convulsion formula and thus interpret the potential mechanism of dispersing wind and relieving convulsion formula reversing drug-resistant epilepsy to verify the hypothesis , and furthermore, to provide a new method for “adding Chinese herbal compound to treat drug -resistant epilepsy.

耐药性癫痫是目前癫痫治疗领域的难点，本项目组在前期临床研究发现以银翘散化裁的疏风止痉方添加治疗耐药性癫痫有一定的疗效，基于银翘散具有调节炎性反应的作用，以及新近研究表明了“癫痫反复发作→NF-κB炎症信号通路激活→P-gp过度表达→形成癫痫耐药”这一主线，据此提出假说：“疏风止痉方逆转耐药性癫痫的可能作用机制是：通过调节癫痫脑内NF-κB炎症信号通路，抑制P-gp的过度表达，提高脑内抗痫药物浓度，从而逆转癫痫耐药，提高临床疗效。”本研究拟以免疫炎症反应为切入点，以癫痫耐药形成机制为主线，筛选出耐药性癫痫模型大鼠，予各组药物干预后，通过对癫痫大鼠后放电阈值及发作情况，脑脊液药物浓度，脑内炎症信号通路中NF-κB蛋白及上下游细胞因子，脑内各区P-gp等指标检测分析，证明疏风止痉方的添加治疗作用，阐释疏风止痉方逆转耐药性癫痫的机制，为中药复方添加治疗耐药性癫痫提供了一个新的思路。

癫痫是多种原因引起的慢性脑疾患，是严重的神经系统疾病，对患者的生活、学习、工作、婚姻及生育等都可能产生很大的不良影响，因此被列入世界卫生组织全球重点防治的五大神经精神疾病之一。癫痫患病率为5%～10%，儿童是癫痫的高发时期，18岁以下儿童占全部癫痫患者的60%以上。近20多年来，尽管不断有新型AEDs问世，但仍有40%的患儿不能得到有效控制、反复发作，成为耐药性癫痫。本项目以癫痫免疫炎症反应为切入点，以“癫痫反复发作→NFκB炎症信号通路激活→P-gp过度表达→形成耐药”为主线，先制作锂-匹罗卡品点燃癫痫大鼠模型，根据痫性发作情况筛选耐丙戊酸钠及卡马西平癫痫大鼠模型，从疏风止痉方对大鼠脑脊液中卡马西平浓度,疏风止痉方对海马NFκB、炎症上下游细胞因子、P-gp的表达来研究疏风止痉方逆转耐药性癫痫的作用及机制。研究结果验证了疏风止痉方可降低NFκB及NFκB炎症信号通路上下游细胞因子的表达，抑制P-gp的表达及转运，提高癫痫大鼠脑内卡马西平浓度，从而减少癫痫发作。这为我们在临床中添加具有调节免疫炎症作用的中药复方治疗耐药性癫痫提供了实验依据。