电针对CPSP大鼠星形胶质细胞介导的Wnt/β-catenin信号通路调控作用

Stroke is a second cause of death and the leading disability in the adult of the world. Central Post-stroke pain (CPSP) is neuropathic pain occurs in the central nervous system after apoplexym, which has ascended to the disease that the mortality rate ranked second in the world and the leading cause of disability in adults. Microglia activation induced central sensitization occurs abnormal pain is the main reason of CPSP. Our previous studies proved that electro acupuncture can reduce the rat models with CPSP’s mechanical pain threshold and heat pain threshold, and can down regulate the expression of β-catenin in brain tissue of rats with CPSP.Thus, we hypothesized that electroacupuncture at "Baihui" and "Zusanli" affecting glial cell differentiation and cell polarity through the Wnt / beta catenin pathway, to maintain the stability of β-catenin in glial cells.This study intends to establish CPSP model to simulate clinical stroke onset by autologous blood injection method, through high frequency electroacupuncture stimulate at "Baihui" and "Zusanli" acupoints in the treatment of CPSP, by observing the changes of intracerebral glial cells after using high frequency electroacupuncture in the treatment of CPSP, to make further detection that glial cell hypertrophy leads to the change of Wnt / β-catenin signaling pathway in order to further elucidate the central regulation mechanismthe of high frequency electro acupuncture for CPSP, and can selective using the inhibitor Dkk1 in Wnt / β-catenin signaling pathway influence the effect of high frequency electro acupuncture for CPSP?This study intends to use the double immunofluorescent labeling techniques, microfluidic analysis technique, enzyme immunoassay and Western blotting method to provide a theoretical basis for the central regulatory mechanism of electro acupuncture in CPSP.

脑卒中跃居为致死率世界第二和主要致残疾病，脑卒中后疼痛（CPSP）是指卒中后中枢神经系统发生的疼痛。胶质细胞激活是导致CPSP主要原因。我们前期证实电针能降低CPSP大鼠的痛阈值且15Hz效果优于其它频率。此外，电针能下调β-catenin在脑组织表达。我们推测电针通过介导Wnt/β-catenin通路调控星形胶质细胞起到镇痛作用。本研究通过自体血注入模拟临床建立CPSP模型，通过电针“百会”和“足三里”观察CPSP大鼠脑星形胶质细胞变化，检测胶质细胞的生物物理特性高通量表征，观察Wnt/β-catenin信号通路的改变以期阐明电针治疗CPSP的中枢机制，而使用Wnt/β-catenin通路抑制剂Dkk-1能否对电针治疗效果产生影响？本研究拟运用免疫荧光双标技术、微流控分析技术、免疫吸附酶联反应和Western blotting方法为电针治疗CPSP的中枢机制提供依据。

脑卒中后疼痛（CPSP）是与卒中病灶直接相关的神经病理性疼痛，前期证实电针能起到缓解CPSP大鼠疼痛的作用，下调β-catenin在脑组织表达，在此基础上，本研究成功建立CPSP大鼠模型，采用不同频率电针观察其镇痛效果，完成了对CPSP模型大鼠脑皮质和海马区不同区域胶质细胞的超微形态学观察，检测了β-catenin/COX-2通路相关因子（COX-2、β-catenin、TGF-β1等）和自噬相关蛋白（LC3B、p62、LAMP-1）的表达，并且完成了应用β-catenin通路特异性抑制剂Dkk-1对结果的验证。综上，本研究发现电针可能是通过调控β-catenin/COX-2信号通路从而降低炎症因子的释放和有效遏制病理性自噬的发生，进而缓解疼痛，进一步揭示了电针对CPSP的中枢调节机制，为电针治疗CPSP提供了理论依据。