芎芷地龙汤对偏头痛模型三叉神经血管系统PKC通路调节作用的研究

In the pathogenesis of migraine, trigeminovascular system is the anatomical basis of pain transmission, At different anatomicallevels，protein kinase C integrates numerous receptor pathways into final effectors that increase excitatory signaling and decrease inhibitory signaling,and then inducing pain occurrence,thus, signaling pathway of protein kinase C plays an important role in the pathophysiology of migraine. This project will Study the ethology of migraine animal,the function level of meningeal vascular of trigeminovascular system, the activation level of protein kinase C and its downstream effector of C-AMP response element binding protein, signal transducer and activator of transcription factor 1 and nuclear factor kappa B In meninges，Trigeminal Ganglion，spinal nucleus of trigeminal nerve in trigeminovascular system,further clarify status of trigeminovascular system and PKC related cell signal transduction in trigeminovascular system after given NO donor,and to elucidate the mechanism of Xiongzhi DiLong Decoction which has expelling wind and removing heat and Promoting blood circulation effect in treatment of migraine.The completion of the project will have important contribution to the biological mechanism of hyperalgesia about the cell signal transduction pathways in migraine which is in pathology condition,and have important theoretical consequences and suitable value in treating migraine with expelling wind and removing heat treatment and Promoting blood circulation treatment,and have an important role in promoting the development of traditional Chinese medicine.

在偏头痛发病过程中，三叉神经血管系统是痛觉传导的解剖基础，在不同的解剖水平，蛋白激酶C集成了众多的受体通路进入可增加兴奋性信号和减少抑制性信号的最终效应器，进而诱发疼痛的产生，蛋白激酶C信号通路在偏头痛的病理生理过程中发挥了重要的作用。本研究通过研究偏头痛模型行为学、三叉神经血管系统脑膜血管功能水平、脑膜、三叉神经节、三叉神经脊束核中蛋白激酶C及其下游效应因子C-AMP反应元件结合蛋白、信号传导及转录激活因子1和细胞核因子κB激活水平，进一步阐明给予NO供体后三叉神经血管系统的激活状态及偏头痛发作的蛋白激酶C相关细胞信号传递过程，并研究祛风清热活血通络方药芎芷地龙汤对其干预的效应机制。课题的完成对从细胞信号传导通路角度阐述偏头痛病理状态下痛觉敏化的生物学机理有重要的理论贡献，对祛风清热活血通络法治疗偏头痛有重要的理论意义和适用价值，对促进祖国医学的发展有重要的作用。

项目的背景.近年研究认为，偏头痛发生相关的解剖结构是三叉神经血管系统。在神经系统的伤害性感受过程，PKC集成了众多的受体通路可增加兴奋性信号和减少抑制性信号的最终效应器，进而诱发疼痛的产生。基因转录是细胞外信号触发神经元长时间变化的关键所在，多种分子或蛋白质，可通过多个激酶激活CREB磷酸化；NF-κB和STAT1依赖的信号转导途径是多种炎性基因转录调控的整合，这些转录因子往往协同调节许多基因的转录激活。.为进一步深入探讨芎芷地龙汤对偏头痛的作用及机理，从PKC及其下游效应因子激活水平方面，进一步阐明其治疗偏头痛的效应机制。.主要研究内容.通过机械痛阈和温度痛阈测定，观察给予NTG后2h动物的痛觉敏化行为，通过对偏头痛动物模型神经肽释放、血管活性物质、PKC活性及其下游效应因子CREB，STAT1和NF-κB表达水平检测，观察模型动物注射NTG后2h血管紊乱水平及PKC介导的细胞信号传导通路，研究芎芷地龙汤对血管紊乱水平及PKC介导的细胞信号传导通路的影响。.重要结果、关键数据.预防给药后3天、7天，芎芷地龙汤均可以提高动物病理状态下的痛阈值，可减弱NO的升高，可升高TXB2降低，缓解6-Keto-PGF1α的降低，升高ET-1含量;减弱NKA的释放;升高CGRP降低水平，预防给药7天效果好于预防给药3天、1天。.芎芷地龙汤预防给药可以减少NTG诱发的三叉神经节、三叉神经脊束核、硬脑膜iNOS、c-fos、CREB1a、CREB1b、STAT1a，升高STAT1b、IL-1β表达，可降低丘脑、三叉神经节Iκ-Bα表达，升高硬脑膜、三叉神经脊束核丘脑Iκ-Bα表达，但以预防给药7天影响明显。.其科学意义.NO 供体 NTG 可诱发动物机械痛阈、温度痛阈降低，并可诱导三叉神经血管系统 PKC 相关的细胞信号传递过程，引起中枢敏化。.芎芷地龙汤能通过调节三叉神经血管反射系统 PKC 相关的细胞信号传递过程，进而减弱因中枢敏化而致痛觉信号的放大和持久，达到预防和治疗偏头痛的作用。