推拿按揉环跳穴对坐骨神经痛大鼠脊髓背角NF-κB p65蛋白的干预作用

Intervention effect of Tuina pressing and kneading the Huantiao(GB30)acupoint on NF-κB p65 protein at spinal cord dorsal horn in sciatica rats

目的:观察推拿按揉环跳穴对坐骨神经慢性压迫损伤(chronic constriction injury,CCI)模型大鼠的镇痛作用,探讨推拿对坐骨神经痛大鼠的镇痛机制。方法:选用32只体重180~220g SPF级别的SD雄性大鼠,随机分成空白组(不予以任何处理)、假手术组(只暴露不结扎坐骨神经)、模型组(结扎坐骨神经)和推拿组(结扎坐骨神经后予以手法干预)。通过结扎大鼠右侧坐骨神经制备CCI模型,于造模第3天开始对推拿组大鼠推拿按揉环跳穴干预,连续干预14 d,观察造模前及造模后第1、3、7、10、14、17天大鼠机械痛域(paw withdrawal threshold,PWT)、热痛阈(paw withdrawal latency,PWL);观察造模前、造模后第1和17天右侧坐骨神经功能指数(sciatic functional index,SFI)的变化;用苏木精伊红(hematoxylin-eosin,HE)染色方法观察坐骨神经形态学的变化;并观察大鼠右侧脊髓背角NF-κB蛋白表达的差异。结果:在造模后,空白组和假手术组的PWT、PWL和SFI差异均无统计学意义(P>0.05),造模后模型组和推拿组的PWT、PWL和SFI显著下降(P<0.01)。在手法干预后,推拿组大鼠的痛阈值上升,在手法干预第8天(即造模第10天),推拿组较模型组PWT显著上升,差异有统计学差异(P<0.01);在手法干预第5天(即造模第7天),推拿组PWL较模型组显著上升,差异有统计学意义(P<0.01);推拿组大鼠痛阈值随着手法干预持续而继续上升。手法干预14天后,推拿组大鼠坐骨神经功能指数显著上升(P<0.01)。与空白组、假手术组比较,模型组大鼠坐骨神经有髓神经纤维排列紊乱,轴索、髓鞘密度不均匀;与模型组比较,推拿组大鼠神经纤维逐渐连续,轴索、髓鞘较模型组均匀。与空白组、假手术组比较,模型组大鼠右侧脊髓背角NF-κB蛋白表达显著增加(P<0.01);与模型组比较,推拿组大鼠右侧脊髓背角NF-κB蛋白表达显著下降(P<0.01)。结论:推拿按揉环跳穴能恢复神经纤维的排列;并通过降低脊髓背角的NF-κB p65蛋白表达来提高CCI模型的PWT、PWL和SFI,从而起到镇痛的作用,并改善大鼠步态。

Objective To observe the analgesic effect of Tuina by pressing and kneading the Huantiao(GB30)acupoint on rats with chronic constriction injury(CCI)and to explore the analgesic mechanism of Tuina on sciatica rats.Methods Thirty-two SPF male SD rats weighing 180 to 220 g were randomly divided into fore groups:blank group(without any treatment),sham group(only exposed without sciatic nerve ligating),model group(sciatic nerve ligating)and Tuina group(manual intervention after lsciatic nerve ligating).The CCI model was prepared by ligating the right sciatic nerve of the rats,on the third day of modeling,the rats in the Tuina group were given pressing and kneading the Huantiao(GB30)point for 14 days,and the changes of paw withdrawal threshold(PWT),paw withdrawal latency(PWL)were measured before and on the 1st,3rd,7th,10th,14th and 17th days after modeling.The changes of sciatic functional index(SFI)were measured before and on the 1st and 17th day after modeling.The morphological changes of the sciatic nerve were observed by hematoxylin-eosin(HE)staining;and the differences in NF-κB protein expression in the right dorsal horn of the spinal cord of rats were detected.Results Following modeling,there was no significant difference in PWT,PWL and SFI between the blank group and the sham group(P>0.05),but the PWT,PWL and SFI of the model group and the Tuina group decreased significantly(P<0.01).After manual intervention,the pain threshold of rats in Tuina group increased.On the 8th day of manual intervention(the 10th day after modeling),PWT in Tuina group increased significantly compared with that in model group(P<0.01).On the 5th day of manual intervention(the 7th day after modeling),the PWL of the massage group was significantly higher than that of the model group(P<0.01).The pain threshold of rats in Tuina group continued to rise with the continuous manipulation intervention.After 14 days of manipulative intervention,the sciatic nerve function index of rats in the Tuina group increased significantly(P<0.01).Compared with the blank group and sham group,the myelinated nerve fibers of sciatic nerve in the model group were disordered and the density of axons and myelin sheath was uneven.Compared with the model group,the nerve fibers of rats in the Tuina group were gradually continuous and the axons and myelin sheath were more uniform than those in the model group.Compared with the blank group and sham group,the expression of NF-κB protein in the right spinal dorsal horn of the model group was significantly increased(P<0.01).Compared with the model group,the expression of NF-κB protein in the right spinal dorsal horn of rats in Tuina group decreased significantly(P<0.01).Conclusions Pressing and kneading the Huantiao(GB30)point restores nerve fiber alignment;and improves the PWT、PWL and SFI in the CCI model by decreasing NF-κB p65 protein expression in the spinal dorsal horn.There fore,Tuina demmstrates an analgesic effect and improves the gait of rats with sciatica.