从脊髓胶质细胞-神经元交互作用探索乌头汤对神经病理性疼痛的镇痛机制

The latest “nerve inflammation theory” views that the action of the activation of glia cells in spinal cord mediating neuronal sensitization, induced by nerve lesions, plays an important role in the pathogenesis of neuropathic pain (NP), and might be a new therapeutic target in the treatment of NP. Wutou decoction, originated from “Synopsis of prescriptions of the golden chamber”, is a classic and has outstanding curative effect in treating pain in clinics. Our research team have previously proved that wutou decoction have new analgesic effect except the known effect on inflammatory pain, while the exact mechanisms is unclear. In this project, we adopt the classic SNL mouse model of NP and combine the non-contact co-cultivation or separate cultivation system of primary microglia/astrocytes and neurons at the same time to study the analgesic effect and characteristics of wutou decoction on NP in different pathological stages by methods such as confocal, calcium imaging, double immunostaining, electron microscope, molecular biology, et al. This project will provide scientific basis for wutou decoction in treating pain in clinics and provide basis for subsequent mechanism exploration. Also, it is helpful for further development of new drugs.

我们前期已研究并初步阐释了来自《金匮要略》“痛痹”要方乌头汤抗类风湿性关节炎的免疫抗炎调节原理。本研究针对乌头汤对神经病理性疼痛（NP）有良好镇痛作用，但机制未明的情况，根据NP是脊髓胶质细胞活化介导的神经炎症的慢性病理过程的认识，从“脊髓胶质细胞-炎症反应-神经元”这一重要调控途径入手，探索乌头汤对不同病理时期NP的镇痛作用、特点及相关机制。研究拟采用NP经典的脊神经结扎（SNL）小鼠模型，通过电镜、激光共聚焦、免疫双标、钙离子荧光成像、WB和Real Time-PCR等多种技术和方法，从形态到功能，局部到系统，多层面、多角度进行研究，以明确乌头汤的NP镇痛作用和特点，并了解初步的作用途径和环节，为后续的机制探索提供依据，也有助于临床的合理运用和进一步的新药研发。

经典名方--乌头汤临床治疗痛痹效果显著，对治疗坐骨神经痛等神经病理性疼痛也取得良好疗效，但其药效特点及相关机制不明确。本研究以经典脊神经结扎模型（SNL）小鼠为研究对象，聚焦于慢性疼痛维持的脊髓中枢，探索了乌头汤对神经病理性疼痛的干预效应及相关机制。发现乌头汤对神经病理性疼痛有良好的干预效应，具体表现为：对SNL小鼠机械痛敏、热辐射痛敏及对机械痛敏基础阈值的改善效应；且无明显的副作用，具体表现为：无耐药性，无明显的毒副作用，不改变动物的痛敏感觉基础阈值。乌头汤对SNL的镇痛效应和抑制脊髓背角星形胶质细胞IL-1R1/TRAF6/JNK信号通路相关，并采用基因芯片结合网络分析的方法发现乌头汤对SNL小鼠的镇痛效应和MAPK、钙离子信号通路、神经受体配基等多个信号通路相关。相关研究深入揭示了乌头汤治疗痛痹的科学内涵，对基于多靶点的新药开发提供了思路，并能以此为典范促进经典名方分子生物学的发展及新药研发，具有重要的现实意义。