齐刺环跳穴干预坐骨神经损伤大鼠海马IL-1β、IL-6、TNF-α及GFAP表达影响的实验研究

Experimental Study on Effect of Triple Needling of Huantiao(GB30)on Expressions of IL-1β,IL-6,TNF-a and GFAP in Hippocampus Japonicus of Rats with Sciatic Nerve Injury

目的 通过观察齐刺环跳穴对坐骨神经损伤大鼠海马白细胞介素-1β(Interleukin-1β,IL-1β)、白细胞介素-6(Interleukin-6,IL-6)、肿瘤坏死因子-α(Tumor Necrosis Factor-alpha, TNF-α)及胶质纤维酸性蛋白(Glial Fibrillary Acidic Protein, GFAP)表达的影响,探讨齐刺环跳穴治疗坐骨神经痛的中枢镇痛机制。方法 60只SD大鼠随机分为假手术组、模型组、齐刺组、单刺组及药物组,每组12只,采用钳夹法制备大鼠坐骨神经损伤模型。造模第2天开始进行针刺及药物干预,连续14 d。观察各组大鼠干预前后坐骨神经功能指数(Sciatic nerve Function Index, SFI)、热缩足反射潜伏期(Paw Withdrawal Latency, PWL)的变化,ELISA法检测海马组织IL-1β、IL-6、TNF-α蛋白表达水平,实时定量PCR法及免疫组化法检测海马组织GFAP表达水平。结果 干预前,与假手术组比较,其余各组大鼠PWL值、SPI值显著降低(P<0.01),与模型组比较,齐刺组干预后大鼠PWL值、SPI值显著改善(P<0.01)。ELISA法、RT-PCR法及免疫组化检测结果显示,模型组、齐刺组、单刺组、药物组IL-1β、IL-6、TNF-α、GFAP表达较假手术组显著升高(P<0.01);与模型组相比,齐刺组、单刺组、药物组IL-1β、IL-6、TNF-α、GFAP表达显著下降,齐刺组表达低于单刺组及药物组(P<0.01)。结论 齐刺环跳穴缓解坐骨神经痛的镇痛机制可能与下调海马炎症因子表达,抑制星形胶质细胞活化,从而降低中枢痛觉敏化程度有关。

Objective To explore the potential analgesic mechanism of triple needling of Huantiao(GB30)in treating neuro-pathic pain caused by sciatic nerve injury by observing the effect of triple needling of Huantiao(GB30)on the expressions of in-terleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-alpha(TNF-α)and glial fibrillary acidic protein(GFAP)in hippocampus of rats with sciatic nerve injury.Methods Sixty SD rats were randomly divided into three groups:sham operation group,model group,triple needling group,single needling group and drug group,with 12 rats in each group.The rat sci-atic nerve injury model was prepared by clamp method.Acupuncture treatment was started on the second day of modeling,once a day,for 14 days.The changes of paw withdrawal latency(PWL)and sciatic nerve function index(SPI)of rats in each group were observed before and after intervention.The hippocampus was taken out to detect the expressions of IL-1β,IL-6 and TNF-αprotein by ELISA,and the expression of GFAP was detected by RT-PCR and immunofluorescence.Results Before treatment,compared with those in the sham operation group,the values of PWL and SPI in each group decreased(P<0.01),and compared with those of the model group,the values of PWL and SPI in the triple needling group significantly improved after treatment(P<0.01).ELISA,RT-PCR and immunofluorescence detection showed that the expressions of IL-1β,IL-6,TNF-αand GFAP in the model group,triple needling group,single needling group and the drug group were significantly higher than those in the sham operation group(P<0.01).Compared with those of the model group,the expressions of IL-1β,IL-6,TNF-αand GFAP in the triple needling group,single needling group and drug group decreased significantly,and the expression in the triple needling group was lower than that in single needling and drug group(P<0.01).Conclusion The analgesic mechanism of triple needling of Huantiao(GB30)in relieving neuropathic pain caused by sciatic nerve injury may be related to down-regulating the expressions of inflammatory factors in hippocampus,inhibiting the activation of astrocytes and reducing the degree of central pain sensitization.