蒙药那如-3通过抑制脊髓胶质细胞活化治疗神经病理性疼痛的机制研究

According to TCM syndrome square measuring principle, the important pathogenesis of neuropathic pain (NP): spinal glial cell activation-driven neuroinflammation, could be attributable to cold “Xie-Ri-Wu-Su” disease category in Mongolian Medicine, which was also verified by the remarkable effect of Mongolian Medicine Naru-3 (NR-3) in clinical treatment of sciatica et al., but the mechanisms remain to be clear. Our previous study found that mouse hyperalgesia thresholds had a cumulative effect and were associated with the reduction of spinal positive glial cells by NR-3 chronic administration in NP model, so we hypothesized that “It is the suppression of glial cell activation in the spinal cord, which caused by chronic administration of NR-3, and the subsequent dispersion of ‘Xie-Ri-Wu-Su’ that lead to the alleviation of central sensitization in NP mouse”. Thus, our project intends to adopt the classic L5 spinal nerve ligation model to observe thermal and mechanical hyperalgesia to clarify the pharmacodynamics of NR-3 on NP mice; and screen the efficacy of target cells by ICH and AimPlex Multiple Immunoassays for Flow; by using glial cells or neurons cultures, glial-glial cells or glial-neurons co-cultures in vitro, and in line with westernblotting, electrophysiological testing, confocal laser scanning techniques to explore the approach, links and targets of NR-3 on the regulation of glial cells. Our results will help explain the multi-targeted mechanisms of NR-3 on NP treatment, guide rational NR-3 use in clinic, and further, for the development of new ethnic medicine, which possesses important significance for NP study and treatment.

根据中医以方测证原理，脊髓胶质细胞活化介导的神经性炎症（神经病理性疼痛 [NP]重要发病机制）可归属于蒙医寒性“协日乌素”病范畴，这也在那如-3治疗坐骨神经痛的显著疗效中得到验证。但其药效机制不明。我们前期研究发现那如-3慢性给药对NP小鼠痛敏阈值有累加效应并与脊髓阳性胶质细胞数目降低相关，因此提出“那如-3存在对胶质细胞活化抑制，进而驱除脊髓‘协日乌素’达到缓解中枢敏化的镇痛机制”假说。项目拟采用经典脊神经结扎模型观察那如-3对NP小鼠机械、热痛敏的影响；IHC筛选药效靶细胞；AimPlex流式高通量多因子检测技术筛选药效相关因子；并配合体外胶质细胞、神经元培养及共培养方法，通过电生理检测、激光共聚焦、WB、PCR等技术多方面探索那如-3对胶质细胞功能调控的途径、环节、靶点；相关研究结果将有助于阐释蒙药那如-3治疗NP的多靶向作用机制，对指导民族药的临床合理用药及进一步研发也具重要意义。

受单靶点药物副作用的限制，如何有效干预神经病理性疼痛（NP）仍是困扰临床医生的棘手问题。基于蒙药那如-3（NR-3）治疗坐骨神经痛等效果明显而机制不明的情况，本研究在成功建立经典脊神经结扎模型（SNL）的基础上开展NR-3药效学研究发现，NR-3具有良好的瞬时镇痛效应及基础痛敏阈值的改善效应，无明显副作用，不改变正常动物痛敏基础阈值，且发现早期给药对基础痛敏阈值的改善效应更明显，提示伴随神经损伤类疾病积极处理原发病的同时应积极早期镇痛；研究还发现SNL模型为“病证结合”的“寒痹”动物模型；机制研究中发现，NR-3瞬时镇痛效应通过阿片受体（δ，μ）激动剂样作用发挥；miRNA基因芯片分析表明，NR-3可能通过调节NP病理状态下的神经免疫系统、内分泌系统、脂质代谢系统、细胞信号转导系统等发挥多方面的药理作用；其对NP早期、维持期基础痛敏阈值的改善效应与上调mmu-miR-7047-5p基因表达调控MMP9/IL-1β、MMP2/IL-1β信号通路进而抑制小胶质细胞及星形胶质细胞活化相关。总之，本研究揭示了NR-3对NP不同病理阶段的药效学特点，发现了一种“寒痹”动物模型，提出了神经损伤应积极早期镇痛的理念，同时探索了NR-3多靶点、多维度干预NP的机制及特点。相关研究结果将有助于揭示民族医药的科学内涵并指导临床实践，对民族医药基础理论的创新发展及基于多靶点、多维度小分子组合物镇痛药物的中药民族药新药研制也意义重大。