基于PKM2-STAT3-炎性因子通路介导小胶质细胞激活探讨电针风池穴对偏头痛大鼠中枢敏化的调控机制

Migraine is a complex neurovascular disorder, touch etc. non-nociceptive stimuli causing pain is a common clinical manifestation of the migrainers, this kind of cutaneous allodynia (CA) is an external symptom of central sensitization, which is the independent risk for headache persistent and the progressive development of chronic migraine. Up to now, there is a lack of effective medication for central sensitization. Central sensitization become a clinical problem to be solved urgently. Acupuncture prevention and treatment of migraine has shown a unique advantage, but the exact mechanism is not yet clear. Studies confirmed that microglial activation are closely related to central sensitization by PKM2-STAT3-inflammatory cytokines pathway, which is a new mechanism of central sensitization. Our previous studies found that electro-acupuncture (EA) at Fengchi can significantly relieve CA and inhibit microglial activation and subsequent inflammatory cytokines release in the trigeminal nucleus caudalis (TNC). So we propose a hypothesis that EA at Fengchi can anti-central sensitization via inhibiting microglial activation by regulating PKM2-STAT3- inflammatory cytokines pathway in migraine. In this study, to explore the efficacy mechanism of acupuncture, we focus on the PKM2-STAT3-inflammatory factor pathway, expecting to provide new scientific evidence for the efficacy mechanism of acupuncture intervention migraine.

偏头痛是一种复杂的神经血管疾病，轻触等非伤害性刺激即可引起皮肤疼痛感是偏头痛患者常见的临床表现，这种皮肤异常性疼痛是中枢敏化的外在征象，是偏头痛迁延持续及向慢性化进展的独立危险因素。目前临床上尚缺乏抗偏头痛中枢敏化的有效药物。针刺干预偏头痛已显示出独特优势，但其疗效机理尚不明确。最新研究发现PKM2-STAT3-炎性因子通路介导小胶质细胞激活参与中枢敏化的形成。申请人前期研究发现电针风池穴能显著缓解偏头痛大鼠皮肤异常性疼痛，抑制三叉神经脊束核尾核（TNC）区小胶质细胞激活及炎性因子释放。结合文献及前期研究结果，我们提出假说“电针风池穴可通过调控PKM2-STAT3-炎性因子通路抑制小胶质细胞激活介导抗偏头痛中枢敏化”。本研究通过反复电刺激大鼠硬脑膜诱导偏头痛中枢敏化模型，PKM2-STAT3-炎性因子通路为研究靶点，探讨电针风池穴抗偏头痛中枢敏化的调控机制，为针刺干预偏头痛提供科学证据。

本研究通过反复电刺激大鼠硬脑膜诱导偏头痛中枢敏化模型，PKM2-STAT3炎性因子通路为研究靶点，探讨电针风池穴抗偏头痛中枢敏化的调控机制。.项目研究结果如下：.（1）行为学水平：电针风池穴可显著改善偏头痛模型大鼠中枢敏化相关行为学；①皮肤异常性疼痛:与模型组相比，电针组大鼠在D4- D8天面部及足底机械痛阈值均显著提高（均p < 0.05）。②Homecage自发性偏头痛样行为：电针组大鼠探索和运动时间显著多于模型组（p < 0.05）；而静止、休息及理毛时间却显著少于模型组（p < 0.05）。.（2）形态学水平：电针风池穴显著抑制三叉神经脊束核尾核（trigeminal nucleus caudalis, TNC）小胶质细胞激活（细胞数明显减少）及PKM2阳性小胶质细胞的数量； TNC 区 PKM2 蛋白与Iba-1免疫荧光双染提示PKM2与Iba-1共定位，电针组TNC 区 PKM2阳性小胶质细胞的数量明显少于模型组。.（3）分子生物学水平：.①电针可显著抑制TNC 区Iba-1、 IL-1β 和 TNF-a 蛋白及 mRNA 表达水平（均p < 0.05）。.②电针可显著抑制TNC 区 PKM2-STAT3 通路蛋白及 mRNA 表达水平（均p < 0.05）。.（4）抑制PKM2以验证针刺的作用：PKM2抑制剂在偏头痛中枢敏化中具有部分类针刺样作用。PKM2抑制剂可显著缓解反复硬脑膜电刺激诱导的面部机械痛（D6及D8天，均p < 0.05 ），同时抑制TNC区小胶质细胞及c-Fos神经元的激活（均p < 0.05）。.（5）拓展实验：针刺可抑制小胶质细-胞神经元交互作用介导抗偏头痛中枢敏化；抑制小胶质细胞激活发挥类针刺样作用。电针风池穴可抑制TNC区神经元上炎性因子受体（IL-1β受体、TNF-α受体）的表达（均p < 0.05）；给予小胶质细胞抑制剂（米诺环素）后，反复硬脑膜电刺激诱导的面部机械痛阈在D4-D8天显著提高（均p < 0.05），TNC区激活的c-Fos神经元数量显著下调（p < 0.05）。.本研究从行为学、形态学及分子生物学水平揭示PKM2-STAT3-炎性因子通路介导的小胶质细胞激活是针刺抗偏头痛中枢敏化的机制之一。拓展研究初步证实提示炎性因子-炎性因子受体介导的小胶质细胞-神经元交互作用是针刺抗偏头痛中枢敏化的机制之一。 .