经TRPV1/ TRPA1 -cGMP信号通路探究推拿对minor CCI模型大鼠背根神经节镇痛启动机制
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北京中医药大学针灸推拿学院

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]国家自然科学(No. 82074573; No. 82274675);北京自然科学(No.7232278)。


Exploring the analgesic initiation mechanism of tuina on the dorsal root ganglion of minor CCI model rats via the TRPV1/TRPA1 -cGMP signaling pathway
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1.School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine;2.School of Acupuncture-Moxibustion and Tuina,Beijing University of Chinese Medicine

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    摘要:

    目的:探究三法三穴推拿手法对minor CCI模型大鼠的镇痛启动机制。方法:将56只SD大鼠随机分为8组,即正常组,假手术组,模型1组,模型2组,推拿1组,推拿2组,推拿1+ TRPA1拮抗剂组,推拿2+ TRPV1拮抗剂组。模型组、推拿组和推拿+拮抗剂组建立minor CCI模型。推拿组和推拿+拮抗剂组在造模后7 d进行1次三法(点法、拨法、揉法)三穴(殷门穴、承山穴、阳陵泉穴)干预,模型组与假手术组进行抓握束缚,正常组不予干预。干预结束后各组先后进行机械缩足反射阈值(MWT)和热缩足反射潜伏期(TWL)测试不同性质痛觉的变化;采用硝酸还原酶法观察各组造模侧DRG中NO含量,ELISA、Western blot、qPCR等方法观察各组造模侧DRG内TRPV1/ TRPA1-NO-cGMP-PKG信号通路蛋白、基因表达的变化情况。结果:与模型组比较,行为学检测显示推拿1组、推拿2组MWT 、TWL均有不同程度的延长;推拿1组、推拿2组、推拿1+ TRPA1拮抗剂组、推拿2+ TRPV1拮抗剂组DRG内TRPV1、 TRPA1、NO、sGCβ、cGMP、PKG1的表达量均明显降低。结论:推拿干预1次后即可有效改善周围神经损伤引起的温度觉、机械痛觉过敏症状;推拿可通过TRPV1/ TRPA1-NO-cGMP-PKG信号通路发挥即刻镇痛作用。

    Abstract:

    Objective: To explore the analgesic initiation mechanism of the three-manipulation and three-acupoint of tuina on rats with minor CCI model. Methods: 56 SD rats were randomly divided into 8 groups: normal group, sham group, model 1 group, model 2 group, tuina 1 group, tuina 2 group, tuina 1 + TRPA1 antagonist group, and tuina 2 + TRPV1 antagonist group. The model group, tuina group, and tuina + antagonist group were established with minor CCI models. The tuina group and tuina + antagonist group received the three-method three-point intervention (point method, dial method, kneading method, Yinmen point, Chengshan point, Yanglingquan point) 7 days after modeling. The model group and sham group were subjected to grasping restraint, and the normal group was not intervened. After the intervention, each group was tested for changes in thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) to detect different types of pain. The nitrate reductase method was used to observe the NO content in the DRG of each group on the modeling side, and ELISA, Western blot, qPCR, and other methods were used to observe the changes in the protein and gene expression of the TRPV1/TRPA1-NO-cGMP-PKG signaling pathway in the DRG of each group on the modeling side. Results: Compared with the model group, behavioral testing showed that the MWT and TWL were prolonged to varying degrees in the tuina 1 group and tuina 2 group. The expression of TRPV1, TRPA1, NO, sGCβ, cGMP, and PKG1 in the DRG of the tuina 1 group, tuina 2 group, tuina 1 + TRPA1 antagonist group, and tuina 2 + TRPV1 antagonist group were significantly decreased. Conclusion: Tuina intervention can effectively improve the symptoms of thermal and mechanical hyperalgesia caused by peripheral nerve injury after one intervention. Tuina can exert immediate analgesic effects through the TRPV1/ TRPA1-NO-cGMP-PKG signaling pathway.

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  • 收稿日期:2023-12-21
  • 最后修改日期:2024-04-23
  • 录用日期:2024-06-28
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