摘要
目的 观察推拿对神经病理性疼痛(NP)大鼠机械缩足反射阈值(PWT)、热缩足反射潜伏期(PWL)、脊髓背角白细胞介素 1β(IL-1β)、白细胞介素 6(IL-6)及c-Fos蛋白表达的影响,探讨推拿疗法缓解NP的作用机制.方法 将36 只SPF级雄性SD大鼠随机分为空白组、坐骨神经慢性压迫性损伤(CCI)组、CCI+推拿组,每组 12 只.采用铬肠线结扎一侧坐骨神经干制作大鼠CCI模型.CCI造模后第 4 天,CCI+推拿组予推拿按揉造模侧"委中"穴,每次 10 min,每日 1 次,持续 14 d.检测造模前及造模后第 4、10、14、17 天各组大鼠PWT、PWL;干预取材后,采用ELISA法检测大鼠造模侧脊髓背角IL-1β、IL-6含量,尼氏染色观察大鼠造模侧脊髓背角神经元组织形态,免疫荧光染色法检测大鼠造模侧脊髓背角c-Fos蛋白表达.结果 与空白组比较,造模后第 4、10、14、17 天CCI组PWT降低(t=27.52、21.80、14.36、16.23,P均＜0.05);与CCI组比较,造模后第 10、14、17 天CCI+推拿组PWT升高(t=-13.58、-6.83、-8.40,P均＜0.05).与空白组比较,造模后第 4、10、14、17 天 CCI组PWL降低(t=16.61、27.22、24.55、27.21,P均＜0.05).与CCI组比较,造模后第 10、14、17 天CCI+推拿组PWL升高(t=-9.35、10.79、-21.86,P均＜0.05).与空白组相比,CCI组大鼠造模侧脊髓背角神经元结构受损,尼氏小体减少.与CCI组相比,CCI+推拿组大鼠造模侧脊髓背角神经元结构受损改善,尼氏小体增加.与空白组比较,CCI组大鼠造模侧脊髓背角组织中IL-1β、IL-6 含量升高(t=-13.97、10.89,P均＜0.05);与CCI组比较,CCI+推拿组大鼠造模侧脊髓背角组织中IL-1β、IL-6 含量降低(t=2.98、6.36,P均＜0.05).与空白组比较,CCI组大鼠造模侧脊髓背角中c-Fos蛋白表达增多(t=-7.97,P＜0.05);与CCI组比较,CCI+推拿组大鼠造模侧脊髓背角中c-Fos蛋白表达减少(t=7.14,P＜0.05).结论 推拿可以有效升高CCI大鼠PWT、PWL,改善脊髓背角神经元组织结构形态,缓解NP,其作机制可能与减少IL-1β、IL-6等炎症因子的表达水平,从而抑制脊髓背角神经元的活化相关.
Abstract
Objective To observe the effect of Tuina on the mechanical paw withdrawal threshold(PWT),thermal paw withdrawal latency(PWL),and the expression of interleukin-1β(IL-1β),interleukin-6(IL-6)and c-Fos in the spinal dorsal horn of neuropathic pain model rats(NP)and to explore the mechanism of Tuina in relieving NP.Methods 36 male SD rats were randomly divided into blank group,chronic construction injury(CCI)group and CCI+Tuina group,with 12 rats in each group.The CCI model was established by ligating the sciatic nerve trunk with chromic catgut.In the CCI+Tuina group,Tuina was applied at ＂Weizhong＂(BL 20)on the operated side for 10 min each time,once a day for 14 days.The PWT and PWL of rats in each group were detected at baseline and Day 4,10,14 and 17 days after modeling.After intervention,the expression of IL-1β and IL-6 in the dorsal horn of the spinal cord were detected by ELISA,nissl staining was used to observe the morphology of neurons in the spinal dorsal horn.and the expression of c-Fos was detected by immunofluorescence staining.Results Compared with the blank group,the PWT of the CCI group decreased on days 4,10,14 and 17(t=27.52,21.80,14.36 and 16.23,all P＜0.05).Compared with the CCI group,PWT in the CCI+Tuina group increased on Days 10,14 and 17(t=-13.58,-6.83,-8.40,all P＜0.05).Compared with the blank group,the PWL of the CCI group decreased on Days 4,10,14 and 17(t=16.61,27.22,24.55 and 27.21,all P＜0.05).Compared with the CCI group,PWL increased in the CCI+Tuina group on Days 10,14 and 17(t=-9.35,10.79,-21.86,all P＜0.05).Compared with the blank group,the structure of neurons in the spinal dorsal hornon of the CCI group was damaged and the number of Nissl bodies reduced.Compared with the CCI group,the damage of neuronal structure in the spinal dorsal horn of the CCI+Tuina group was improved,and Nissl bodies increased.Compared with the blank group,the expression of IL-1β and IL-6 in the spinal dorsal hornon of the CCI group increased(t=-13.97,10.89,all P＜0.05).Compared with the CCI group,the expression of IL-1β and IL-6 in the spinal dorsal horn decreased in the CCI+Tuina group(t=2.98,6.36,all P＜0.05).Compared with the blank group,the expression of c-Fos in the spinal dorsal hornincreased in the CCI group(t=-7.97,P＜0.05).Compared with the CCI group,the expression of c-Fos in thespinal dorsal horn decreased in the CCI+Tuina group(t=7.14,P＜0.05).Conclusions Tuina at ＂Weizhong＂(BL 20)can effectively increase PWT and PWL of CCI rats,improve the structure and morphology of neurons in the spinal dorsal horn,and alleviate NP.The mechanism may be related to reducing the expression levels of inflammatory factors such as IL-1β and IL-6,thereby inhibiting the activation of spinal dorsal horn neurons.
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