基于红外测温的火针“温通”效应改善脊髓损伤作用靶点研究

Target study of fire needle "Wentong" effect in improving spinal cord injury based on infrared temperature measurement

关键词:  脊髓损伤  火针  热休克蛋白  靶点  神经细胞凋亡

Key words:  spinal cord injury  fire needle  heat shock protein  target  neuronal apoptosis

摘要:

[目的] 在红外测温的基础上，观察火针疗法对HSP60蛋白的影响，探讨火针疗法“温通”效应改善脊髓损伤（SCI）的作用靶点。[方法] 将120只SD大鼠随机分为假手术组、模型组、毫针组、艾灸组、火针组，每组分为1、3、7和14 d共4个时点。采用改良的Allen’s法制备SCI模型，对毫针组、艾灸组和火针组分别采用相应的干预方法，相应时点进行Basso Beattie Bresnahan （BBB）行为功能评分、红外测温、免疫组化检测及TUNEL染色检测。[结果] 1）行为学评分显示，与模型组相比，火针组在3、7和14 d时评分均明显升高，差异具有统计学意义（P<0.05）；与毫针组和艾灸组相比，火针组在14 d时评分明显升高，差异具有统计学意义（P<0.05）。2）红外测温结果显示，火针组在火针干预5 min后温度呈缓慢上升趋势，升高0.5℃左右，且升温过程持续至90 min。3）免疫组化结果显示，假手术组在各个时点HSP60的阳性表达相对稳定；而模型组在1 d时HSP60的阳性表达低于假手术组；与模型组各个时点相比，毫针组、艾灸组的HSP60阳性表达均增高；与毫针组和艾灸组相比，火针组HSP60阳性表达升高，且在7 d和14 d时升高最明显。4）TUNEL染色结果显示，假手术组在各个时点均无或存在少量神经细胞凋亡情况；模型组在1 d时凋亡细胞明显增多；3 d时毫针组、艾灸组和火针组神经细胞凋亡情况少于模型组，持续至7 d和14 d，其中火针组在14 d时仅可见少量神经细胞凋亡。[结论] 火针疗法可升高SCI大鼠脊髓损伤局部温度，减轻神经元凋亡，进而改善其神经功能缺损症状，而其“温通”效应的作用靶点可能与HSP60有关。

Abstract:

[Objective] To observe the effect of fireneedle therapy on HSP60 on the basis of infrared temperature measurement,and to explore the target of fire needle therapy on improving spinal cord injury. [Methods] One hundred and twenty SD rats were randomly divided into sham operation group,model group,filiform needle group,moxibustion group and fire needle group,each group was divided into 4 time points which were 1,3,7 and 14 d. The SCI models were built by modified Allen's method,and the corresponding intervention methods were used in the filiform needle group,moxibustion group,and fire needle group,respectively. Basso Beattie Bresnahan (BBB) score,infrared temperature measurement,immunohistochemical detection and TUNEL staining detection were performed at the corresponding time points. [Results] 1) BBB scores showed that compared with the model group,scores of the fire needle group were significantly higher at 3,7 and 14 d,with statistical significance (P<0.05). Compared with the filiform needle group and moxibustion group,the scores in the fire needle group at 3 and 14 d were significantly higher (P<0.05). 2) Infrared temperature measurement results showed that the temperature of the fire needle group showed a slow rising trend,to increase about 0.5 ℃,and the heating process lasted for 90 minutes after the fire needle intervention. 3) Immunohistochemical results showed that the positive expression of HSP60 was relatively stable at each time point in sham operation group,while the positive expression of HSP60 in the model group was lower than that in the sham operation group on 1 d. Compared with the model group at each time point,the positive expression of HSP60 in the filiform needle group and moxibustion group increased. Compared with the filiform needle group and the moxibustion group,the positive expression of HSP60 in the fire needle group increased,and the increase was most obvious at 7 and 14 d. 4) TUNEL staining results showed that there was no or a small amount of neuronal apoptosis at each time point in the sham operation group,and apoptotic cells increased significantly in the model group on 1 d. At 3 d,the apoptosis of neurons in the filiform needle group,moxibustion group,and fire needle group was less than that in the model group,which lasted until 7 and 14 d,and only a few nerve cells in the fire needle group were apoptotic at 14 d. [Conclusion] Fire needle could increase the local temperature of SCI rats,decrease neuron apoptosis toimprove the symptoms of neurological impairment,and the target of "Wentong" effect might be related to HSP60.