电针预处理经miR-34a-5p调节IL-1β/COX2/PGE2炎症通路预防偏头痛机制研究

Migraine is a neurovascular disorder with gastrointestinal and visual symptoms that affects about 15% of people worldwide. Acupuncture is effective for migraine prophylaxis and for management of acute migraine attacks, without the adverse secondary effects associated with medications. Activation of the trigeminovascular system plays an essential role in inducing pain during a migraine attack. IL-1β stimulates COX-2 dependent PGE2 synthesis and CGRP release in rat trigeminal ganglia cells. The IL-1β/COX2/PGE2 inflammatory pathway contributes to activation of the trigeminovascular system. Electroacupuncture pretreatment inhibits IL-1β/COX2/PGE2 inflammatory pathway in the trigeminal ganglion of rats with migraine, and electroacupuncture plays protective roles in some diseases. Migraine attacks are associated with an acute upregulation in miR-34a-5p expression. It had be found that miR-34a/SIRT1 modulates expression of IL-1β. There is a need to investigate whether electroacupuncture can prevent migraine attacks by increasing SIRT1 expression via down-regulating expression of miR-34a-5p in the trigeminal ganglion, which inhibits IL-1β/COX2/PGE2 inflammatory pathway. Firstly, the effect of miR-34a-5p/SIRT1 on IL-1β/COX2/PGE2 inflammatory pathway in the trigeminal ganglion should be investigated in vitro. Secondly, we would testify the hypothesis whether electroacupuncture can prevent migraine attacks through miR-34a-5p/SIRT1 regulating IL-1β/COX2/PGE2 inflammatory pathway in the trigeminal ganglion in vivo.

针刺预防偏头痛疗效确切。IL-1β/COX2/PGE2炎症通路参与三叉神经血管系统激活。前期研究显示电针抑制偏头痛大鼠三叉神经节IL-1β/COX2/PGE2炎症通路。偏头痛发作时miR-34a-5p表达上调。miR-34a/SIRT1调节IL-1β表达。因此，我们提出“电针预处理抑制miR-34a-5p表达，上调SIRT1表达，进而抑制IL-1β/COX2/PGE2炎症通路，减少CGRP 释放，预防偏头痛发作”的工作假说。首先通过体外实验证实miR-34a-5p/SIRT1是否调控三叉神经节IL-1β/COX2/PGE2炎症通路，再以偏头痛大鼠模型为研究对象，验证电针预处理是否经miR-34a-5p/SIRT1抑制三叉神经节IL-1β/COX2/PGE2炎症通路，从而抑制三叉神经血管系统激活，论证上述假设是否存在。

偏头痛是一种常见的、复杂的、伴有感觉改变的致残性神经系统疾病，占据神经系统疾病经济负担的五成以上。针灸防治偏头痛历史悠久，疗效可靠，世界卫生组织已将“偏头痛”列为针灸治疗优势病种之一，但其作用机制尚未完全清楚，有待进一步探究。本课题依据项目计划书的安排如期进行，深入探讨电针治疗偏头痛潜在的分子机制。首先以原代三叉神经节细胞为研究对象，通过过表达转染或沉默技术、RT-PCR、Western blot及ELISA等实验技术，发现miR-34a-5p通过抑制三叉神经节神经元中SIRT1的表达上调IL-1β/COX2/PGE2炎症途径，诱导细胞凋亡并增加CGRP释放。随后以电刺激三叉神经节大鼠偏头痛模型为研究对象，电针处理为干预手段，证实电针可抑制偏头痛大鼠三叉神经节miR-34a-5p表达,增加SIRT1表达,下调IL-1β/COX2炎性反应信号,减少PGE2合成,进而减少三叉神经末梢释放CGRP。本研究为偏头痛治疗开创了新的思路和理论依据，为针刺治疗在偏头痛中的应用提供了进一步的理论支持。在本项目支持下迄今共发表2篇科学论文，其中SCI论文1篇，北大中文核心论文1篇，另有1篇科学论文已接收（待刊）。