Academy of Medical Engineering and Translational Medicine, Tianjin University, Tianjin, 300072, China.
Research Center of Experimental Acupuncture Science, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China.
Chin J Integr Med. 2021 Dec;27(12):933-939. doi: 10.1007/s11655-020-3435-6. Epub 2020 Nov 2.
To identify the prominent molecular signaling in acupoints and explore their roles in initiating the analgesia effect of manual acupuncture (MA).
A three-step study was conducted, the experiment 1 was a genome-wide analysis of the tissue at acupoint Zusanli (ST 36), including 12 Wistar rats which were divided into control, control+MA1, and control+MA7 groups. In the experiment 2, the paw withdrawal latency (PWL), immunohistochemistry and Western blot analysis of phospho-nuclear factor kappa B (NFκB) p65 (p-p65), phospho-NFκB p50 (p-p50) at ST 36 were performed on rats of saline, saline+MA, and complete Freund's adjuvant (CFA)+MA groups (n=6). In experiment 3, 24 rats were divided into saline+DMSO, CFA+DMSO, CFA+DMSO+MA, and CFA+BAY 11-7082+MA groups, the PWL and immunofluorescence assay of NFκB p65 at ST 36 was conducted.
(1) The gene: inhibitor of NFκB (Nfkbia), interleukin-1β (Il1b), interleukin-6 (Il6), chemokine c-x-c motif ligand 1 (Cxcl1), monocyte chemoattractant protein-1 (MCP-1/Ccl2) expressions in the control+MA7 group were significantly increased (P<0.05 or P<0.01), and the expression of NFκB p65 (Rela), NFκB p50 (Nfkb1) were increased in the control+MA7 group (P<0.05). (2) CFA+MA groups showed increased PWL from day 1 to 7 (P<0.01 vs. CFA), and the Western blot results were consistent with immunohistochemistry, the expression of NFκB p-p65 and NFκB p-p50 were significantly increased in the MA-related groups compared with control and CFA groups (P<0.05). (3) Compared with the CFA+DMSO+MA group, the PWL of the CFA+ BAY 11-7082+MA group decreased significantly and continued until day 5 and 7 (P<0.05 and P<0.01, respectively), and the NFκB p65 expression of CFA+BAY 11-7082+MA was significantly reduced compared with CFA+DMSO+MA (P<0.01).
Local NFκB signaling cascade in acupoint caused by MA is an important step in initiating the analgesic effect, which would provide new evidence for the initiation of MA-effect and improve the understanding of the scientific basis of acupuncture analgesia.
确定穴位中突出的分子信号，并探讨它们在引发手动针刺（MA）镇痛作用中的作用。
进行了三步研究，实验 1 是对穴位足三里（ST 36）组织的全基因组分析，包括分为对照组、对照组+MA1 组和对照组+MA7 组的 12 只 Wistar 大鼠。在实验 2 中，对盐水、盐水+MA 和完全弗氏佐剂（CFA）+MA 组的大鼠进行了足底退缩潜伏期（PWL）、磷酸核因子 kappa B（NFκB）p65（p-p65）的免疫组织化学和 Western blot 分析（n=6）。在实验 3 中，将 24 只大鼠分为盐水+DMSO、CFA+DMSO、CFA+DMSO+MA 和 CFA+BAY 11-7082+MA 组，进行了 ST 36 处 NFκB p65 的 PWL 和免疫荧光分析。
（1）对照组+MA7 组中抑制 NFκB（Nfkbia）、白细胞介素-1β（Il1b）、白细胞介素-6（Il6）、趋化因子 c-x-c 基序配体 1（Cxcl1）和单核细胞趋化蛋白-1（MCP-1/Ccl2）的基因表达明显增加（P<0.05 或 P<0.01），并且 NFκB p65（Rela）和 NFκB p50（Nfkb1）的表达在对照组+MA7 组中增加（P<0.05）。（2）CFA+MA 组从第 1 天到第 7 天 PWL 明显增加（与 CFA 相比，P<0.01），Western blot 结果与免疫组织化学结果一致，与对照组和 CFA 组相比，MA 相关组中 NFκB p-p65 和 NFκB p-p50 的表达明显增加（P<0.05）。（3）与 CFA+DMSO+MA 组相比，CFA+BAY 11-7082+MA 组的 PWL 显著降低，并且一直持续到第 5 天和第 7 天（分别为 P<0.05 和 P<0.01），并且 CFA+BAY 11-7082+MA 组的 NFκB p65 表达与 CFA+DMSO+MA 组相比明显降低（P<0.01）。
MA 引起的穴位局部 NFκB 信号级联反应是启动镇痛作用的重要步骤，为 MA 效应的启动提供了新的证据，并提高了对针刺镇痛科学基础的理解。
