Research Article

Combined Auricular Electrostimulation and Sunshi Dingtong Decoction Relieve Primary Dysmenorrhea With Qi Stagnation and Blood Stasis in Rats

DOI:

10.3791/70584

⸱

June 22nd, 2026

 ,  ,  ,  ,  , 
1Department of Gynecology, Fuzhou Hospital of Traditional Chinese Medicine, 2Department of Oncology, Fuzhou Hospital of Traditional Chinese Medicine

* These authors contributed equally

In This Article

Summary

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Transcutaneous auricular electrostimulation combined with Sunshi Dingtong decoction alleviated uterine pathological injury and inflammation-related changes in a rat model of primary dysmenorrhea with qi stagnation and blood stasis, accompanied by altered mRNA expression of PPAR-γ2 and COX-2.

Abstract

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Primary dysmenorrhea with a qi stagnation and blood stasis pattern is associated with uterine inflammation and pain, and current pharmacologic therapies may cause adverse effects. This study evaluated the combined therapeutic effects of transcutaneous auricular point electrostimulation (TEAP) and Sunshi Dingtong decoction (SDD) in a rat model of qi stagnation and blood stasis primary dysmenorrhea. Thirty-six female Sprague–Dawley rats were randomly allocated to the blank, model, SDD, TEAP, TEAP plus SDD, and ibuprofen groups. The model was induced using estradiol benzoate sensitization, epinephrine hydrochloride combined stress stimulation, and oxytocin administration. Rats received normal saline, SDD, ibuprofen, or TEAP at defined auricular points for seven days. Uterine morphology was examined by hematoxylin and eosin staining. Uterine levels of IL-1β, IL-6, TNF-α, PGF2α, and PGE2 were measured by ELISA, and uterine PPAR-γ2 and COX-2 mRNA expression was quantified by real-time PCR. Model rats showed uterine epithelial degeneration, neutrophil infiltration, increased IL-1β, IL-6, TNF-α, PGF2α, and COX-2 mRNA expression, and decreased PGE2 and PPAR-γ2 mRNA expression. TEAP or SDD alone partially reversed these pathological and molecular changes, whereas combined TEAP plus SDD produced more pronounced improvements, with effects comparable to those observed in the ibuprofen group. These findings suggest that TEAP combined with SDD alleviates uterine pathological injury and inflammation-related changes in qi stagnation and blood stasis primary dysmenorrhea (QSBSPD) rats, and that these effects are associated with the regulation of PPAR-γ2 and COX-2 mRNA expression.

Introduction

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Primary dysmenorrhea (PD), alternatively termed functional menstrual pain, is characterized by the occurrence of dysmenorrhea without any identifiable organic pathology within reproductive organs1. Female patients experiencing severe dysmenorrhea frequently find themselves unable to perform daily tasks, which significantly impacts their quality of life and necessitates pharmacological intervention for symptom relief2. Extant literature indicates that it is associated with various factors, including prostaglandins (PGs), estrogen, progesterone, vasopressin, oxytocin (OT), the immune system, neuroendocrine effects, and alt....

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Protocol

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Ethical statement

All animal procedures were reviewed and approved by Fujian Ambry Biotechnology Laboratory Animal Ethics Committee approval number:IACUC  FJABR2024031202). Perform all animal experiments in accordance with the approved institutional animal care and use protocol.

SDD and ibuprofen preparation

SDD consists of Shixiao San (9 g), Xiangfu (9 g), Moyao (6 g), Guizhi (9 g), Weilingxian (12 g), Shijianchuan (20 g), Yanhusuo (12 g), Danggui (9 g), Liujinu (15 g), and Xiaohuixiang (5 g). Shixiao San consists of Puhuang (4.5 g) and Wulingzhi....

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Results

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Screening of the estrous cycle in rats

The results of Wright-Giemsa staining showed that the vaginal smears in preoestrus predominantly exhibited nucleated epithelial cells, with a small amount of keratinocytes. The nuclei of these epithelial cells appeared lightly stained, enlarged, and showed signs of dissolution. In oestrus, the smears were characterized by a predominance of large, deciduous keratinized epithelial cells. In postestrus, there was a substantial presence of ke.......

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Discussion

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PD is the lower abdominal pain that manifests before and during menstruation without any identifiable organic damage in the genital or pelvic regions1. According to the survey, approximately 45%–90% of women globally experience dysmenorrhea, with 10% to 25% suffering from severe dysmenorrhea13. Therefore, it is imperative to identify safer and more effective interventions for PD and to further elucidate their underlying mechanisms.

Currentl.......

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Disclosures

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The authors have no conflicts of interest to declare.

Acknowledgements

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This research is supported by Fuzhou Health Science and Technology Innovation Platform-“Fuzhou Traditional Chinese Medicine Heritage Innovation Platform” (grant no. 2021-S-wp3) and Fujian Natural Science Foundation project-“Based on PPAR-gamma-2 /COX-2 signaling pathway, the mechanism of percutaneous electrical stimulation combined with Sunshi Dingtong Decoction in qi stagnation and blood stasis primary dysmenorrhea via regulation” (grant no. 2022J01121509).

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Materials

List of materials used in this article
NameCompanyCatalog NumberComments
0.9% sodium chloride solutionHeilongjiang Qiqin Animal Health Products Co., Ltd.240314A03Tissue rinsing and sample preparation
1× phosphate-buffered saline (PBS)BiosharpBL601ATissue rinsing, ELISA homogenization, and staining procedures
Acoustic and light stimulation systemCustom-built in animal facilityCustom-builtCombined stress stimulation during QSBSPD model construction
Adhesive microscope slidesJiangsu Citotest Labware Manufacturing Co., Ltd.80312-3161Preparation of paraffin sections and vaginal smears
Automated digital slide scannerGuangzhou Guangying Cell Technology Co., Ltd.GCell-60Digital imaging of HE-stained sections
Automatic tissue dehydratorHubei Xiaogan Kuohai Medical Technology Co., Ltd.KH-TSTissue dehydration before paraffin embedding
Biological microscopeNikonE200MVObservation of vaginal smears and stained tissue sections
CO2 gas cylinder and euthanasia chamberFuzhou Hospital of Traditional Chinese Medicine Animal CenterInstitutional CO2 systemAnimal euthanasia
Cold plate for embedding stationHubei Xiaogan Kuohai Medical Technology Co., Ltd.KH-BLParaffin embedding
Cryogenic grinderShanghai Jingxin Industrial Development Co., Ltd.TISS-24Tissue homogenization
Danggui (Radix Angelicae Sinensis)Guangdong Yifang Pharmaceutical Co., Ltd.A1070343SDD ingredient; concentrated granules
DEPC-treated waterMeilunbioMA0018qRT-PCR reagent
Electronic acupuncture treatment instrumentSuzhou Medical Appliance Factory Co., Ltd.SDZ-IITEAP stimulation and electrical stimulation
Electronic analytical balanceSartorius Balance Beijing Co., Ltd.BS 224SSample and reagent weighing
EosinEpredia7111HE staining
Epinephrine hydrochlorideTargetMol119091QSBSPD model construction
Estradiol benzoateSigma-AldrichE8515QSBSPD model construction
EthanolXilong Scientific Co., Ltd.240403A1Tissue processing and dehydration
Guizhi (Ramulus Cinnamomi)Guangdong Yifang Pharmaceutical Co., Ltd.A2060533SDD ingredient; concentrated granules
HematoxylinEpredia7211HE staining
Ibuprofen suspensionShanghai Johnson & Johnson Pharmaceutical Co., Ltd.H20000359Positive control drug; 20 mg/mL
Liujinu (Herba Artemisiae Anomalae)Guangdong Yifang Pharmaceutical Co., Ltd.A210A413SDD ingredient; concentrated granules
Low-speed mini centrifugeHunan Xiangyi Laboratory Instrument Development Co., Ltd.WTL-4KSample preparation
Micro refrigerated centrifugeBeckman CoulterMicrofuge 22RRNA extraction and tissue homogenate preparation
Microplate readerGuangdong Danli Technology Co., Ltd.K3ELISA absorbance measurement at 450 nm
Microscope cover slipsJiangsu Citotest Labware Manufacturing Co., Ltd.80340-1630Microscopy slide preparation
Mold incubatorShanghai Lichen Bangxi Instrument Technology Co., Ltd.W0704Incubation during staining or assay procedures
Moyao (Myrrh)Guangdong Yifang Pharmaceutical Co., Ltd.1107853SDD ingredient; concentrated granules
Neutral balsamMeilunbioMB9899Mounting of stained tissue sections
NovoScript Plus All-in-one 1st Strand cDNA Synthesis SuperMixNovoproteinE047-01AReverse transcription
NovoStart SYBR qPCR SuperMix PlusNovoproteinE096-01BqRT-PCR amplification
Oxytocin injectionAnhui Fengyuan Pharmaceutical Co., Ltd.220906-4QSBSPD model induction
Paraffin waxHualing20220918Paraffin embedding
PCR systemBio-RadPTC-100PCR-related procedures
Puhuang (Pollen Typhae)Guangdong Yifang Pharmaceutical Co., Ltd.A2092143SDD ingredient; concentrated granules; component of Shixiao San
qRT-PCR primers for PPAR-γ2, COX-2, and GAPDHSangon Biotech (Shanghai) Co., Ltd.Custom synthesisqRT-PCR primers
Rat IL-1β ELISA kitMeimian BiotechnologyMM-0047R2ELISA kit
Rat IL-6 ELISA kitMeimian BiotechnologyMM-0190R2ELISA kit
Rat PGE2 ELISA kitMeimian BiotechnologyMM-0068R2ELISA kit
Rat PGF2α ELISA kitMeimian BiotechnologyMM-0230R2ELISA kit
Rat restraint cageCustom-built in animal facilityCustom-builtRestraint stress during QSBSPD model construction
Rat TNF-α ELISA kitMeimian BiotechnologyMM-0180R2ELISA kit
Real-time PCR systemApplied Biosystems7300qRT-PCR analysis
RNAiso PlusTaKaRa9109Total RNA extraction
Rotary microtomeThermo Fisher ScientificHM325Paraffin sectioning
Shijianchuan (Herba Salviae Chinensis)Guangdong Yifang Pharmaceutical Co., Ltd.A3052113SDD ingredient; concentrated granules
Sprague–Dawley ratsBeijing Vital River Laboratory Animal Technology Co., Ltd.SCXK (Jing) 2019-0010Animal model construction
Tissue embedding cassetteJiangsu Citotest Labware Manufacturing Co., Ltd.23033Paraffin sectioning consumable
Tissue embedding machineHubei Xiaogan Kuohai Medical Technology Co., Ltd.KH-BLParaffin embedding
Tissue flotation bath and slide warmerHubei Xiaogan Kuohai Medical Technology Co., Ltd.KH-P2Paraffin section mounting
Ultra-low temperature freezerZhongke Meiling Cryogenics Co., Ltd.DW-HL340Sample storage
Ultramicro UV-Vis spectrophotometerBeijing BioTeke CorporationND5000RNA concentration and purity measurement
Ultrapure water systemSichuan Delishi Technology Co., Ltd.SP-D2-V-10LSPreparation of ultrapure water
Weilingxian (Radix et Rhizoma Clematidis Chinensis)Guangdong Yifang Pharmaceutical Co., Ltd.A2030543SDD ingredient; concentrated granules
Wright-Giemsa stainSolarbioG3005Estrous cycle screening
Wulingzhi (Faeces Trogopterori)Guangdong Yifang Pharmaceutical Co., Ltd.A2082223SDD ingredient; concentrated granules; component of Shixiao San
Xiangfu (Rhizoma Cyperi)Guangdong Yifang Pharmaceutical Co., Ltd.A2069413SDD ingredient; concentrated granules
Xiaohuixiang (Fructus Foeniculi)Guangdong Yifang Pharmaceutical Co., Ltd.A1125153SDD ingredient; concentrated granules
XyleneXilong Scientific Co., Ltd.240124A2Dewaxing and tissue clearing
Yanhusuo (Rhizoma Corydalis Yanhusuo)Guangdong Yifang Pharmaceutical Co., Ltd.A2032983SDD ingredient; concentrated granules

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Tags

Primary DysmenorrheaQi StagnationBlood StasisAuricular ElectrostimulationSunshi Dingtong DecoctionUterine InflammationHematoxylin Eosin StainingReal Time PCRELISA AssayCOX 2 Expression

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