针灸对CCI大鼠NOX-ROS-NLRP3通路镇痛作用及抗炎机制的研究 - 龙华针灸

Objective: To observe the effects of acupuncture and moxibustion therapy on pain relief in sciatica rats and to explore the mechanism of its anti-inflammatory effect.

Methods: SPF grade 4-6-week-old Kunming rats were randomly divided into 5 groups including a blank group, sham-operated group, model group, acupuncture, and moxibustion (AnM) group, and positive group. A total of 10 rats were included in each group. The model group, the AnM group, and the positive group were prepared by ligating the left sciatic nerve. AnM group was used for acupuncture and moxibustion therapy intervention, and the positive group was rendered to quick-acting sciatica pills once a day for 7 days (3 courses of treatment). The blank group, sham-operated group, and model group were not treated. The changes in thermal and mechanical pain thresholds were observed before and after the operation, and the morphological changes of the dorsal horn of the spinal cord in the lumbosacral region of the rats in each group were observed by HE staining after the courses of treatment finished. The contents of IL-1β, IL-6, IL-18, and TNF-α were measured by ELISA and the expressions of NOX1, NOX2, NOX4, and NLRP3 genes were detected by RT-qPCR while the protein expressions of NOX1, NOX2, NOX4 and NLRP3 were analyzed by Western blotting.

Results: The AnM and positive group showed a significant increase in thermal and mechanical pain thresholds after treatment, while there was no significant change in the model group. As compared to the control group, the contents of IL- 1β, IL-6, IL-18, and TNF-α, as well as the relative expressions of NOX1, NOX2, NOX4, and NLRP3 genes were significantly increased in the model group (P < 0.05 or P < 0.01). As compared to the model group, the contents of IL-1β, IL-6, IL-18, and TNF-α, as well as the relative expressions of NOX1, NOX2, NOX4, and NLRP3 genes significantly decreased in the AnM and positive groups (P < 0.05 or P < 0.01). The pathological changes of inflammatory infiltration of tissue cells in the dorsal horn of the lumbosacral spinal cord were slowed in the AnM group.

Conclusion: Acupuncture and moxibustion therapy have a positive effect on pain relief and anti-inflammatory effects in CCI sciatica rats, which may point to the regulation of NOX1, NOX2, NOX4, and NLRP3 expressions, and inhibition of ROS.

Keywords: Acupuncture; Inflammation; Mechanism; Moxibustion therapy; Sciatica.

摘要目的：观察针灸对坐骨神经痛大鼠的镇痛作用，探讨其抗炎作用的机制。方法：SPF 4～6周龄昆明大鼠随机分为5组，分别为空白组、假手术组、模型组、针灸组和阳性组。每组共10只大鼠。模型组、AnM组和阳性组通过结扎左坐骨神经制备。AnM组采用针灸治疗干预，阳性组给予速效坐骨神经痛丸，每日1次，疗程7天（3个疗程）。空白组、假手术组和模型组均未治疗。观察手术前后热痛阈值和机械痛阈值的变化，并在疗程结束后通过HE染色观察各组大鼠腰骶部脊髓背角的形态学变化。ELISA法检测IL-1β、IL-6、IL-18和TNF-α的含量，RT-qPCR法检测NOX1、NOX2、NOX4和NLRP3基因的表达，Western印迹法分析NOX1、NOx 2、NOx 4和NLRP3。结果：AnM和阳性组治疗后热痛阈值和机械痛阈值显著升高，而模型组没有显著变化。与对照组相比，模型组IL-1β、IL-6、IL-18和TNF-α的含量以及NOX1、NOX2、NOX4和NLRP3基因的相对表达均显著增加（P<0.05或P<0.01），AnM组和阳性组NLRP3基因表达显著降低（P＜0.05或P＜0.01）。结论：针灸对CCI坐骨神经痛大鼠具有积极的镇痛和抗炎作用，可能与调节NOX1、NOX2、NOX4和NLRP3的表达以及抑制ROS有关。关键词：针灸；发炎机械装置艾灸疗法；坐骨神经痛。