GSK-3β调控三叉神经节CX3CL1/CX3CR1途径参与偏头痛的机制研究

CX3CL1/CX3CR1 pathway mediated neuron-glia crosstalk plays an important role in neuropathic pain. However, involvement of CX3CL1/CX3CR1 pathway is still remaining unknown in migraine. Our previous study demonstrated increased expression of CX3CL1/ CX3CR1 in trigeminal ganglion (TG) in rats with migraine, suggesting that CX3CL1/CX3CR1 pathway may involve in the occurrence of migraine. We also found that glycogen synthase kinase 3β (GSK-3β) was highly activated in TG neurons of rats with migraine, and GSK-3β inhibitors could prevent CX3CL1/CX3CR1 overexpression, which indicates that GSK-3β might regulate CX3CL1/CX3CR1 pathway via the crosstalk between neurons and satellite glial cells in TG in migraine. The current study will include a series of in vivo and in vitro experiments, such as real-time quantitative PCR, Western Blot, co-culture of neurons and satellite glial cells, co-immunoprecipitation, RNAi, etc. The outcome of current project may provide a novel drug target for migraine’s treatment.

CX3CL1/CX3CR1途径介导神经元-胶质细胞Cross-Talk在神经病理性疼痛中发挥重要作用，但CX3CL1/CX3CR1途径是否参与偏头痛过程却知之甚少。我们前期研究发现偏头痛大鼠三叉神经节（TG）表达CX3CL1/CX3CR1增加，提示CX3CL1/CX3CR1途径参与了偏头痛过程，同时发现TG神经元糖原合酶激酶3β（GSK-3β）磷酸化激活，GSK-3β抑制剂可以抑制CX3CL1/CX3CR1过表达。因此推测，GSK-3β可能调控CX3CL1/CX3CR1途径介导TG神经元与卫星胶质细胞Cross-Talk参与偏头痛过程。本课题将通过体内和体外实验，利用实时定量PCR、Western Blot、免疫共沉淀、细胞共培养、RNAi 等手段验证上述推测并阐明其机制，探讨介导TG神经元与卫星胶质细胞Cross-Talk参与偏头痛发生发展的关键因素，为偏头痛靶向药物研发提供新的策略。

CX3CL1/CX3CR1途径介导神经元-胶质细胞Cross-Talk在神经病理性疼痛中发挥重要作用，但CX3CL1/CX3CR1途径是否参与偏头痛过程却知之甚少。本研究发现，硝酸甘油型偏头痛模型大鼠三叉神经节组织CX3CL1/ CX3CR1表达增强，GSK-3β抑制剂能够降低硝酸甘油诱导的三叉神经节中的CX3CL1/ CX3CR1过度表达，其中对主要表达于卫星胶质细胞的CX3CR1蛋白下调最为明显。体外实验发现，CX3CL1主要表达于三叉神经节神经元，SNAP处理后CX3CL1表达明显增强，应用GSK-3β抑制剂，三叉神经节神经元的CX3CL1表达下降，CX3CR1主要定位于卫星胶质细胞，表达趋势与CX3CL1相似。抑制NF-κB活性，能够影响上游蛋白GSK 3β激活，说明NF-κB信号对GSK 3β具有反馈调节作用。应用NF-κB信号途径激活剂联合GSK-3β抑制剂，三叉神经节神经元CX3CL1蛋白表达增加，而CX3CR1的表达影响不明显。本研究表明，GSK-3β能够调控CX3CL1/CX3CR1途径介导三叉神经节神经元与卫星胶质细胞间交互作用参与偏头痛的周围敏化过程，为偏头痛防治提供了新的研究靶点。