益气活血通络方对糖尿病神经病理性疼痛大鼠脊髓星形胶质细胞活性及JNK信号通路的影响

Effects of Yiqi Huoxue Tongluo Formula on astrocyte activity and JNK signaling pathway in spinal cord of diabetic neuropathic pain rats

目的:探讨益气活血通络方防治糖尿病神经病理性疼痛(DNP)的作用机制。方法:高脂喂养联合单次小剂量注射STZ制备2型糖尿病大鼠,通过检测机械痛阈(MWT)和热缩足反应时间(TWL)判定DNP模型成功。Western blot检测脊髓JNK、p-JNK、GFAP蛋白表达;ELISA检测脊髓IL-1β、IL-10和TNF-α的表达;免疫荧光法检测脊髓星形胶质细胞活性。结果:与模型组比较,益气活血通络方各剂量组大鼠MWT、TWL均显著改善(P<0.01);大鼠脊髓JNK、p-JNK、GFAP蛋白表达均显著降低(P<0.01,P<0.05);大鼠脊髓炎性因子IL-1β、IL-10、TNF-α均显著降低(P<0.01,P<0.05);且DNP大鼠脊髓星形胶质细胞活性受到显著抑制(P<0.01)。结论:益气活血通络方对DNP大鼠具有明显的防治作用,其作用机制可能是通过抑制JNK信号通路,调节脊髓星形胶质细胞活性,从而减轻神经炎症反应导致的中枢敏化来实现的。

Objective: To explore the mechanism of Yiqi Huoxue Tongluo Formula in preventing and treating diabetic neuropathic pain(DNP) in vivo experiment. Methods: Rats model with type 2 diabetes mellitus were induced by highfat feeding combined with low dose of streptozotocin(STZ) for once. Mechanical withdrawal threshold(MWT) and thermal withdrawal latency(TWL) results determined the model of DNP built successfully. Western blot detected the protein expression of JNK, p-JNK, GFAP in spinal cord tissue;ELISA detected the expression of IL-1β, IL-10 and TNF-α in spinal cord tissue;immunofluorescence detected spinal cord astrocytes activity. Results: Compared with the model group, the MWT and TWL of the three dose of Yiqi Huoxue Tongluo Formula groups rats were significantly improved(P<0.01), the protein expression levels of JNK, p-JNK and GFAP in rat spinal cord tissues were significantly decreased(P<0.01, P<0.05), the expression levels of inflammatory factors IL-1β, IL-10 and TNF-α in the spinal cord of rats were significantly decreased(P<0.01, P<0.05);meanwhile, the activity of spinal cord astrocytes in DNP rats was significantly inhibited(P<0.01). Conclusion: Yiqi Huoxue Tongluo Formula have significant prevention and treatment effect on DNP rats, and its mechanism might be achieved by inhibiting the JNK signal pathway and regulating spinal astrocyte activity, thereby reducing central sensitization caused by neuroinflammatory reactions.