Efficacy and Mechanism of Shuanghua Drink in Treating Primary Dysmenorrhea Based on COX-2/NF-κB Signaling Pathway
10.13422/j.cnki.syfjx.20251225
- VernacularTitle:基于COX-2/NF-κB信号通路探讨双花饮治疗原发性痛经的药效及作用机制
- Author:
Yuncheng MA
1
;
Yuanyuan SHI
1
;
Zhen LIU
2
;
Yuxi WANG
1
;
Yuan TIAN
2
;
Qian LI
2
;
Xiaozhu WANG
2
;
Cheng HE
3
;
Wenhui XU
3
;
Weiling WANG
3
;
Jian GAO
3
;
Ting WANG
2
Author Information
1. School of Life Sciences， Beijing University of Chinese Medicine， Beijing 100029， China
2. School of Chinese Materia Medica， Beijing University of Chinese Medicine， Beijing 100029， China
3. Beijing Research Institute of Traditional Medicine， Beijing University of Chinese Medicine， Beijing 100029， China
- Publication Type:Journal Article
- Keywords:
Shuanghua drink;
primary dysmenorrhea;
cyclooxygenase-2 / nuclear factor-κB （COX-2 / NF-κB） signaling pathway;
prostaglandin;
inducible nitric oxide synthase （ iNOS ）
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2025;31(12):72-80
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo evaluate the efficacy of Shuanghua drink in treating primary dysmenorrhea in the rat model and explore its mechanism of action. MethodsAn oxytocin-induced writhing mouse model was established to evaluate the analgesic effect of Shuanghua drink. Forty-eight non-pregnant female institute of cancer research （ICR） mice were randomly divided into six groups， including a blank group， a model group， an ibuprofen group （85.00 mg·kg-1）， a low-dose group of Shuanghua drink （7.14 mL·kg-1）， a medium-dose group of Shuanghua drink （14.28 mL·kg-1）， and a high-dose group of Shuanghua drink （28.57 mL·kg-1）. Each group consisted of eight mice. All treatment groups received daily intragastric administration at corresponding doses for 10 consecutive days. One hour after the final administration， 2 U of oxytocin was intraperitoneally injected per mouse. The writhing latency and number of writhing within 20 minutes were recorded. A primary dysmenorrhea rat model was established by using estradiol benzoate and oxytocin to evaluate the inhibitory effect of Shuanghua drink on the contraction of uterine smooth muscle. Forty-eight non-pregnant female Sprague-Dawley （SD） rats were divided into six groups， including a blank group， a model group， an ibuprofen group （51.00 mg·kg-1）， a low-dose group of Shuanghua drink （4.28 mL·kg-1）， a medium-dose group of Shuanghua drink （8.57 mL·kg-1）， and a high-dose group of Shuanghua drink （17.10 mL·kg-1）. Each group consisted of eight rats. Rats received subcutaneous injections of estradiol benzoate for 10 consecutive days to enhance uterine sensitivity. On the eleventh day， oxytocin （2 U/rat） was intraperitoneally administered to induce abnormal uterine contractions for establishing the primary dysmenorrhea model. All treatment groups received daily intragastric administration from the second day of modeling for 10 days. The effects of Shuanghua drink were evaluated by using parameters including uterine motility and the variation rate of uterine motility. The mechanism of action was investigated in rats with primary dysmenorrhea. The content of prostaglandin F2α （PGF2α）， prostaglandin E2 （PGE2）， thromboxane B2 （TXB2）， prostacyclin metabolite （6-keto-PGF1α）， and β-endorphin （β-EP） in uterine tissue of rats was detected by using enzyme-linked immunosorbent assay （ELISA）. The changes in the content of nitric oxide （NO） and inducible nitric oxide synthase （iNOS） were analyzed via colorimetric assay. Western blot was performed to determine the content of phosphorylated inhibitor of kappa B kinase beta （p-IKKβ）/IKKβ， phosphorylated inhibitor of kappa B alpha （p-IκBα）， IκBα， phosphorylated p65 （p-p65）， p65， and cyclooxygenase-2 （COX-2） proteins in uterine tissue of rats. ResultsIn the oxytocin-induced writhing mouse model， the model group exhibited significantly shortened writhing latency and increased writhing frequency compared to the control group （P<0.01）. Both the ibuprofen group and the high-dose group of Shuanghua drink displayed prolonged writhing latency （P<0.05）， while the ibuprofen group and the low-dose， medium-dose， and high-dose groups of Shuanghua drink exhibited reduced writhing frequency （P<0.01）. In the primary dysmenorrhea rat model， the uterine motility and its variation rate in the model group were significantly higher than those in the blank group （P<0.01）. These parameters were markedly suppressed by ibuprofen and Shuanghua drink at all tested doses （P<0.01）. For the mechanism of action， the model group showed significantly increased PGF2α/PGE2， TXB2/6-keto-PGF1α， NO， and iNOS in uterine tissue （P<0.05， P<0.01） and significantly decreased β-EP （P<0.01）. These parameters were significantly attenuated in the ibuprofen group and the low-dose， medium-dose， and high-dose groups of Shuanghua drink. The PGF2α/PGE2 （P<0.01）， TXB2/6-keto-PGF1α （P<0.01）， NO （medium-dose group P<0.05）， and iNOS （P<0.01） were reduced， and the β-EP （medium-dose group P<0.05） was up-regulated. Compared to the model group， the ibuprofen group and medium-dose group of Shuanghua drink showed significantly increased content of β-EP in the serum of rats （P<0.05）. Compared to the blank group， the model group showed significantly elevated expressions of COX-2， p-IKKβ/IKKβ， p-IκBα/IκBα， and p-p65/p65 proteins （P<0.01） and significantly reduced anti-inflammatory protein IκBα （P<0.05）. Compared to the model group， the ibuprofen group and the low-dose， medium-dose， and high-dose groups of Shuanghua drink showed significantly reduced expressions of COX-2 （P<0.01）， p-IKKβ/IKKβ （P<0.01）， p-IκBα/IκBα （P<0.05， P<0.01）， and p-p65/p65（P<0.01） and up-regulated expression of IκBα protein （P<0.05， P<0.01）. ConclusionShuanghua drink effectively alleviates primary dysmenorrhea through analgesia and suppression of abnormal contractions of uterine smooth muscle. Its mechanism may be mediated by reduced levels of PGF2α/PGE2， TXB2/6-keto-PGF1α， iNOS， and NO， elevated β-EP level， and inhibited COX-2/NF-κB signaling pathway.
