火针疗法“温通效应”改善脊髓损伤急性期炎性微环境的机制研究

To study the mechanism of “warming effect” of fire needling therapy in improving the inflammatory microenvironment in the acute stage of spinal cord injury

关键词:  火针  温通效应  脊髓损伤  炎性微环境  针刺

Key words:  fire needle  warming effect  spinal cord injury  inflammatory microenvironment  acupuncture needle

摘要:

目的] 探讨火针疗法改善急性脊髓损伤炎性微环境的作用机制。[方法] 将24只雌性SD大鼠随机分为空白组、假手术组、造模组、火针组，每组6只。采用改良的Allen’s法制备急性脊髓损伤大鼠模型。假手术组切除大鼠T 9、T 10的棘突及部分椎板后不予撞击；造模组造模后不予干预；火针组造模后运用贺氏火针点刺T7、T8、T11、T12棘突下及左右旁开各0.5 cm处，共12个穴位。红外成像技术测量各组大鼠干预前、干预后即刻、干预后10、20、30 min脊髓损伤部位的温度变化；蛋白免疫印迹（Western blot）法检测大鼠脊髓组织中Toll样受体4（TLR4）、髓样分化因子88（MyD88）、核转录因子-κB（NF-κB）的蛋白相对表达水平；酶联免疫吸附法（ELISA）检测血清中肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、白细胞介素-4（IL-4）、白细胞介素-6（IL-6）、白细胞介素-10（IL-10）的含量水平。[结果] 1）红外成像结果显示，火针组在干预后即刻温度明显升高，在治疗后10 min温度下降，随后逐渐趋于稳定。2）Western blot结果显示：与造模组相比，火针组大鼠脊髓组织TLR4相对表达量降低，差异具有统计学意义（P＜0.05），MyD88、p-NF-κB相对表达量降低，但两者差异无统计学意义（P＞0.05）。3）血清ELISA结果：与造模组相比，火针组大鼠血清TNF-α含量降低，差异具有统计学意义（P＜0.05）；与造模组相比，火针组大鼠血清IL-1β、IL-6含量降低，两者差异无统计学意义（P＞0.05）；与造模组相比，火针组大鼠血清IL-4、IL10的含量升高，差异具有统计学意义（P＜0.05）。[结论] 火针疗法“温通效应”可以介导TLR4/MyD88/NF-κB通路调控促炎/抗炎平衡进而改善脊髓损伤急性期炎性微环境。

Abstract:

[Objective] To explore the mechanism of fire needling therapy in improving the inflammatory microenvironment of acute spinal cord injury.[Methods] 24 female SD rats were randomly divided into blank group,sham operation group,model group and fire needling group,with 6 rats in each group. The SCI model was established by modified Allen's method. In the sham-operation group,T9 and T10 spinous processes and partial laminae were removed without intervention. The model group was not intervened after modeling. After modeling,the rats in the fire needling group were stimulated with fire needling at T7,T8,T11,T12 0.5cm below the spinous process and at the left and right lateral points,a total of 12 acupoints. Infrared imaging technology was used to measure the temperature changes of rats before intervention,immediately after intervention,10,20 and 30 min after intervention. The protein expression levels of Toll-like receptors 4(TLR4),myeloid differentiation primary factor88(MyD88) and nuclear factor-kappa B(NF-κB) in spinal cord tissues were detected by Western blot. The levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-4(IL-4),interleukin-6(IL-6) and interleukin-10(IL-10) in serum were detected by enzyme-linked immunosorbent assay.[Results] 1) Infrared imaging results showed that the temperature of the fire needling group increased significantly immediately after intervention,decreased 10min after treatment,and then gradually stabilized. 2) Western blot results showed that compared with the model group,the relative expression of TLR4 in the spinal cord tissue of the fire needling group was decreased(P<0.05),and the relative expressions of MyD88 and p-Nf-κb were decreased,but the differences were not statistically significant(P>0.05). 3) Elisa results:Compared with the model group,the content of serum TNF-α in the fire needling group was decreased(P<0.05). Compared with the model group,the serum levels of IL-1β and IL-6 in the fire needling group were decreased,and the difference was not statistically significant(P>0.05). Compared with the model group,the serum levels of IL-4 and IL10 in the fire needling group were increased(P<0.05).[Conclusion] The "warming effect" of fire needling therapy can regulate the pro-inflammatory/anti-inflammatory balance through TLR4/MyD88/NF-κB pathway to improve the inflammatory microenvironment in the acute stage of spinal cord injury.