Analgesic effect of "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique of acupuncture on primary dysmenorrhea based on NOD1/RIP2/NF-κB signaling pathway in the rats

JIN Xu; ZHANG Yanlin; CHANG Boya; REN Jia; HAO Jianheng; CAO Yuxia; WANG Haijun; JI Laixi

doi:10.13703/j.0255-2930.20240331-k0001

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Analgesic effect of "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique of acupuncture on primary dysmenorrhea based on NOD1/RIP2/NF-κB signaling pathway in the rats

Study on Mechanism | 更新时间：2025-02-14

- Analgesic effect of "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique of acupuncture on primary dysmenorrhea based on NOD1/RIP2/NF-κB signaling pathway in the rats
- Chinese Acupuncture & Moxibustion Vol. 45, Issue 2, Pages: 209-216(2025)
- 作者机构：
  
  1.山西中医药大学第二临床学院，晋中　030619
  2.天津中医药大学针灸推拿学院
  3.山西中医药大学附属针灸推拿医院，太原　030012
- 作者简介：
- 基金信息：
- DOI：10.13703/j.0255-2930.20240331-k0001
  CLC：
- Received：31 March 2024，
  
  Online First：25 November 2024，
  
  Published：12 February 2025
- 稿件说明：
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JIN Xu, ZHANG Yanlin, CHANG Boya, et al. Analgesic effect of "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique of acupuncture on primary dysmenorrhea based on NOD1/RIP2/NF-κB signaling pathway in the rats[J]. Chinese Acupuncture & Moxibustion, 2025, 45(2): 209-216.
DOI：

JIN Xu, ZHANG Yanlin, CHANG Boya, et al. Analgesic effect of "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique of acupuncture on primary dysmenorrhea based on NOD1/RIP2/NF-κB signaling pathway in the rats[J]. Chinese Acupuncture & Moxibustion, 2025, 45(2): 209-216. DOI： 10.13703/j.0255-2930.20240331-k0001.

摘要

目的：

观察“秩边透水道”针法对原发性痛经（PD）大鼠核苷酸结合寡聚化结构域1（NOD1）/受体相互作用蛋白2（RIP2）/核转录因子κB（NF-κB）信号通路相关蛋白和促炎细胞因子表达的影响，探讨其缓解PD疼痛的可能机制。

方法：

将30只SPF级雌性SD大鼠（动情周期正常）随机分为空白组、模型组和针刺组，每组10只。模型组和针刺组大鼠采用苯甲酸雌二醇联合缩宫素腹腔注射法制备PD模型，针刺组大鼠在造模同时予“秩边透水道”针法干预，每次20 min，每天1次，连续10 d。于第11天观察各组大鼠腹腔注射缩宫素注射液30 min内扭体反应（扭体潜伏期、扭体次数、扭体评分）；HE染色法观察大鼠子宫组织形态；ELISA法检测大鼠血清前列腺素E

（PGE

）、前列腺素F

2α

（PGF

2α

）、白细胞介素（IL）-1β、IL-18、环氧合酶-2（COX-2）、肿瘤坏死因子-α（TNF-α）含量；Western blot法检测各组大鼠子宫组织NOD1、RIP2、NF-κB p65、磷酸化核转录因子κB p65（p-NF-κB p65）蛋白表达；实时荧光定量PCR法检测各组大鼠子宫组织NOD1、RIP2、NF-κB p65 mRNA表达。

结果：

与空白组比较，模型组大鼠扭体潜伏期延长（

0.01），扭体次数增加及扭体评分升高（

0.01）；子宫内膜上皮细胞大量变性、坏死，子宫内膜严重水肿且大范围脱落，伴有中性粒细胞浸润；血清PGE

含量降低（

0.01），血清PGF

2α

、IL-1β、IL-18、COX-2、TNF-α含量升高（

0.01）；子宫组织NOD1、RIP2、NF-κB p65、p-NF-κB p65蛋白表达及NOD1、RIP2、NF-κB p65 mRNA表达升高（

0.01）。与模型组比较，针刺组大鼠扭体潜伏期延长（

0.01），扭体次数减少及扭体评分降低（

0.01）；子宫内膜上皮组织少量变性、坏死，子宫内膜轻度水肿，少有中性粒细胞浸润；血清PGE

含量升高（

0.01），血清PGF

2α

、IL-1β、IL-18、COX-2、TNF-α含量降低（

0.01）；子宫组织NOD1、RIP2、NF-κB p65、p-NF-κB p65蛋白表达及NOD1、RIP2、NF-κB p65 mRNA表达降低（

0.05，

0.01）。

结论：

“秩边透水道”针法能够缓解PD大鼠的疼痛症状，其作用机制可能与抑制PD大鼠子宫组织中NOD1/RIP2/NF-κB信号通路的活性，进而减轻其炎性反应有关。

Abstract

Objective

To observe the effect of "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique on the relative protein expression of the signaling pathway of nucleotide-binding oligomerization domain-containing protein 1 (NOD1)/ receptor-interacting protein 2 (RIP2)/nuclear factor kappa-B (NF-κB) and the expression of proinflammatory cytokines in the rats with primary dysmenorrhea (PD)

so as to explore the underlying mechanism of this acupuncture technique for pain alleviation in PD.

Methods

Thirty female SD rats of SPF grade with normal estrous cycle were randomized into a blank group

a model group and an acupuncture group

10 rats in each one. Using the intraperitoneal injection with estradiol benzoate combined with oxytocin

PD model was prepared in the model group and the acupuncture group. In the acupuncture group

during model preparation

the intervention with "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique was delivered simultaneously

20 min each time

once daily for consecutive 10 days. On day 11

within 30 min after the intraperitoneal injection with oxytocin

the writhing reaction (latency

frequency and score) was recorded; the morphology of uterine tissue was observed with HE staining

the contents of prostaglandin E

(PGE

)

prostaglandin F

2α

(PGF

2α

)

interleukin (IL)-1β

IL-18

cyclooxygenase-2 (COX-2)

and tumor necrosis factor-α(TNF-α) in the serum were detected using ELISA method; the relative protein expression of NOD1

RIP2

NF-κB p65

phosphorylation-NF-κB p65 (p-NF-κB p65) was detected in the uterine tissue using Western blot method; and the mRNA expression of NOD1

RIP2 and NF-κB p65 was detected with the quantitative real-time PCR employed.

Results

Compared with the blank group

in the model group

the writhing latency was prolonged (

0.01)

the writhing frequency and score increased (

0.01) in the rats; the endometrial epithelial cells showed massive degeneration and ne

crosis

with severe endometrial edema and widespread shedding

combined with neutrophil infiltration; the serum PGE

content was dropped (

0.01)

while those of PGF

2α

IL-1β

IL-18

COX-2

and TNF-α elevated (

0.01); the protein expression of NOD1

RIP2

NF-κB p65 and p-NF-κB p65

and the mRNA expression of NOD1

RIP2 and NF-κB p65 in uterine tissue increased (

0.01). In comparison with the model group

in the acupuncture group

the writhing latency was prolonged (

0.01)

the writhing frequency and score were reduced (

0.01) in the rats; there was less degeneration and necrosis of endometrial epithelial cells

with mild endometrial edema and very little neutrophil infiltration; the serum PGE

content increased (

0.01)

while those of PGF

2α

IL-1β

IL-18

COX-2

and TNF-α decreased (

0.01); the protein expression of NOD1

RIP2

NF-κB p65 and p-NF-κB p65 and the mRNA expression of NOD1

RIP2 and NF-κB p65 in uterine tissue were dropped (

0.05

0.01).

Conclusion

"Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique can alleviate the pain symptom of PD rats

and its action mechanism may be related to inhibiting the active expression of NOD1/RIP2/NF-κB signaling pathway in the uterine tissue

thereby reducing the inflammatory response.

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