从相关GPCRs参与痛觉调制探讨疏肝调神针法防治偏头痛的作用机制

Migraine is a common clinical primary headache，which the incidence is increasing year by year. The acupunture therapy by dispersing the stagnated liver-qi and regulating the spirit is one of the clinical experience of Shan Qiuhua who is a famous TCM professor in China. Earlier clinical and experimental studies found that acupuncture or bloodletting could effectively reduce the frequence，relieve the pain，regulate the related vasoactive substances.The acupuncture method by dispersing the stagnated liver-qi and regulating the spirit play a good analgesic effect. However，the mechanism is still not clear. In this study，we use the migraine model rats which copied by hypodermic injection nitroglycerin as the research object，to observe the contents of some related vasoactive substances and inflammatory factors，as well as the expressing of GPCRs in central nervous system and GPCRs mediated signal transduction pathways of migraine rats with the related GPCRs as breakthrough points and the trigeminovascular system as the main line. Then，we will further observe the effects of acupuncture therapy on the expressing of GPCRs to discuss the information transfer process of the migraine and central regulation mechanism for migraine, in hope that to study the mechanism of acupuncture therapy by multi-faceted, multi-target, multi-channel methods，and to provide scientific evidence for clinic better.

偏头痛是临床常见的原发性头痛，其发病率逐年增加。疏肝调神针法是全国名中医单秋华教授临床经验之一，前期临床及实验研究发现针刺、放血等疗法治疗偏头痛，可有效减少发作次数，缓解疼痛程度，对相关血管活性物质具有一定调控作用，发挥良好的镇痛作用，但其作用机制尚不明确。本研究以硝酸甘油型偏头痛模型大鼠为研究对象，以相关GPCRs为切入点，以三叉神经血管系统为主线，观察偏头痛模型大鼠相关神经递质、炎性因子的含量变化，以及相关 GPCRs在中枢的表达和GPCRs介导的信号转导通路，进一步观察针刺对其表达的影响，从而深入探讨偏头痛的信息传递过程及针刺对偏头痛的中枢调控机制，以期从多方位、多靶点、多途径等方面研究疏肝调神针法治疗偏头痛的镇痛镇静作用机理，更好的为临床提供科学佐证。

本研究参照经典硝酸甘油造模法，复制偏头痛大鼠模型。对模型大鼠及不同针刺处理后的不同时间段进行行为学评分，探讨疏肝调神针法对模型大鼠的镇痛镇静作用。对各种处理方法后的大鼠，保存组织标本。采用ELISA检测方法，观察针刺对偏头痛大鼠血清相关血管活性物质和炎性介质含量的变化。应用免疫组化、RT-PCR、Western Blot法，观察大鼠三叉神经脊束核和中脑相关GPCRs的表达情况，探讨针刺的防治偏头痛的中枢机制。结果显示，疏肝调神预防组、疏肝调神治疗组和普通针刺组均可改善偏头痛大鼠的行为学症状，具有预防和治疗作用，且疏肝调神治疗组优势更明显。造模后，血清中5-HT含量较正常组降低，SP、CGRP含量升高，提示5-HT、SP和CGRP均参与了偏头痛的发生。针刺后，与模型组比较，疏肝调神预防组、疏肝调神治疗组和普通针刺组5-HT含量升高，SP、CGRP含量均降低，说明针刺干预可调节偏头痛大鼠血管活性物质水平，尤其在调节5-HT、SP方面，疏肝调神治疗组优于普通针刺组。针刺组 IL-1β、TNF-α、ICAM-1 、COX-1、NF-kB等的含量均明显低于模型组，而IL-6的含量却高于模型组，表明疏肝调神针法能降低偏头痛大鼠血清致炎因子IL-1β、TNF-α、ICAM -1 、COX-1、NF-kB等的含量，升高抗炎因子IL-6的含量。同时，针刺对偏头痛大鼠三叉神经脊束核和中脑中相关CGRPs及c-fos、c-jun的蛋白和mRNA表达均有一定的上调或者下调。结论：疏肝调神针法可能通过调节炎性因子的含量，起到抗炎作用，从而调节血管的舒缩以改善症状。针刺通过调节相关CGRPs在三叉神经血管系统通路的表达，从而进一步激活G蛋白通路，起到中枢调节作用，而且疏肝调神针法的调节作用更为显著。