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Herbs-Partitioned Moxibustion on the Navel in a Rat Model of Primary Dysmenorrhea with Cold Coagulation and Blood Stasis
* These authors contributed equally
In This Article
Summary
Here, we describe in detail the operation procedures and precautions to be taken during herbs-partitioned moxibustion on the navel in the treatment of primary dysmenorrhea in rats with cold coagulation and blood stasis, which can significantly improve the writhing reaction of dysmenorrhea rats and alleviate pain.
Abstract
Primary dysmenorrhea (PDM) refers to the occurrence of spasmodic colicky pain in the lower abdomen without any obvious pelvic pathology, often accompanied by other systemic symptoms, which significantly reduces the quality of life of women with PDM, 45%-95% of menstrual women are affected by it. Despite the high incidence of PDM, it is often not well treated and is ignored by relevant researchers and women themselves. Herbs-partitioned moxibustion (HPM) is a characteristic external therapy in traditional Chinese medicine (TCM), which is widely used to treat diseases in China; it treats diseases by applying a Chinese herbal formula on the patient's acupoints and then igniting moxa sticks on the herbal formula for moxibustion therapy. The primary effectiveness of the herbs utilized in this study is meridian activation and warmth, which encourages qi circulation to relieve pain. Moxibustion's heat stimulation could hasten the infusion of the herbs into the human body. Consequently, HPM, which combines acupoint, herbal, and heat stimulations, is appropriate for treating gynecological disorders, particularly functional diseases such as PDM. This paper provides a detailed description of the procedures and precautions for HPM at the Shenque (CV8) acupoint in female experimental rats, which 0provides an experimental basis for better promoting the application and development of HPM on the navel in PDM.
Introduction
Primary dysmenorrhea (PDM), also referred to as functional dysmenorrhea, is one of the most prevalent gynecological diseases affecting adolescent girls and menstruating young women1. It is characterized by side effects such as anguish and uterine compressions (spasms), together with inflammatory problems in the abdomen before and after menstruation without any evidence of pelvic pathology2. Lower abdominal cramps that start 8-72 h after the commencement of menstruation, peak in the first few days of increased menstrual flow3,4, and sometimes spread to the back an....
Protocol
This experimental protocol was approved by the Experimental Animal Management and Ethics Review Committee of Zhejiang University of Traditional Chinese Medicine. All experimental operations on animals comply with the experimental animal welfare ethics and animal experimental safety regulations. The study used adult female Wistar rats aged 8-10 weeks from the Laboratory Animal Center of Zhejiang Chinese Medicine University. Prior to the experiment, rats were fed an adaptive diet for 1 week in a controlled environment (tem.......
Representative Results
In this study, estradiol benzoate in combination with oxytocin was used to induce the PDM model of cold coagulation and blood stasis type to observe the effect of HPM navel therapy. First, by observing the rat's writhing behavior, determine whether the PDM rat with cold coagulation and blood stasis was successfully modeled, and evaluate the rat's pain behavior, modeling, and treatment on the uterus through WS, WLs, and uterine index. From the 6th to the 10th day of modeling, HPM treatment wa.......
Discussion
At present, there are various modeling methods for PDM with cold coagulation and blood stasis type, but the main method is estradiol benzoate combined with oxytocin for dysmenorrhea modeling, which will be different on this basis. In this study, the whole-body freezing method combined with estradiol benzoate and oxytocin was used for modeling. The temperature of the whole body freezing method was -20 °C. In the previous pre-experiment, it was found that when the temperature was −25 °C, the rats would have.......
Acknowledgements
This work was supported by the Zhejiang Provincial Natural Science Foundation, Exploring Program Q (LQ20H270012), Zhejiang Traditional Chinese Medicine Technology Plan (NO.2020ZB124), the Research Project of Zhejiang Chinese Medical University (NO.2021JKZKTS056B), and the Hospital-Level Project of the Third Affiliated Hospital of Zhejiang Chinese Medical University (NO.ZS22ZA01). We are grateful to the Experimental Animal Center of Zhejiang Chinese Medical University and the Key Laboratory of Acupuncture and Neurology of Zhejiang Province for providing us with the experimental conditions.
....Materials
| Name | Company | Catalog Number | Comments |
| 0.9% physiological saline | Hangzhou Gaosheng Biotechnology Co., Ltd. | SLYSZ1A | For injecting the control group |
| 1 mL disposable sterile syringe | Changzhou Yuekang Medical Equipment Co., Ltd. | 3000-0515-14 | For drug injection |
| 3 mL (extended) Pasteur pipette | Beijing Lanjieke Technology Co., Ltd. | YCKJ-HC-007245 | For mounting |
| 75% disinfection alcohol | Hangzhou OUTOP Biotechnology Co., Ltd. | N/A | For sterilizing the treatment site |
| Adhesive-coated glass slide | Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd. | 188105W | For HE staining |
| Biochemical incubator | Shanghai Boxun Industrial Co., Ltd. Medical Equipment Factory | N/A | For tissue dewaxing in HE staining |
| Cloth material | In-house preparation | N/A | Pouch to hold rat |
| Digital manager slice scanning system | Hamamatsu | C13210-01 | For HE slice scanning |
| Disposable latex gloves | Guangzhou Lige Technology Co., Ltd. | LG10-102-1I | For use during operation |
| Estradiol benzoate | Hefei Xinkexin Animal Pharmaceutical Co., Ltd. | N/A | For establishing dysmenorrhea model |
| Excelsior AS Fully automatic dehydrator | Thermo Scientific | Excelsior AS | For HE staining tissue dehydration |
| Female Wistar rats | Laboratory Animal Center of Zhejiang Chinese Medicine University | N/A | 220–250 g |
| Flour | In-house preparation | N/A | For herbs-partitioned moxibustion treatment |
| Forcep | Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd. | lsnz | For specimen collection |
| Formaldehyde | Shanghai Lingfeng Chemical Reagent Co., Ltd. | 441244-1KG | For tissue fixation |
| Gemini AS slide stainer | Thermo Scientific | Gemini AS | Staining machine |
| Hemostat | Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd. | lsnz | For specimen collection |
| HistoCore | Leica ( Arcadia H ) | 14039357258 | For HE staining tissue embedding |
| Medical cotton ball sterile | The Third Affiliated Hospital of Zhejiang Chinese Medical University | N/A | For sterilize the tratment site |
| Methylene blue staining solution (0.1%) | Beijing Honghu United Chemical Products Co., Ltd. | G1300 | For screening estrous cycle |
| Microdissection Shears | Hangzhou Gaosheng Biotechnology Co., Ltd. | XWJ12A | For specimen collection |
| Microscope cover glass (24 mm x 50 mm) | Hangzhou Zhengbo Biotechnology Co., Ltd. | 10212450C | For HE staining |
| Microtome | Leica Microsystems | RM2255 | For tissue sectioning |
| Midea freezer | Hefei Midea Refrigerator Co., Ltd. | N/A | For establishing dysmenorrhea model with cold coagulation and blood stasis |
| Molar forceps | Hangzhou Zhengbo Biotechnology Co., Ltd. | YGJD-11 | For specimen collection |
| Mounting medium | Hangzhou Gaosheng Biotechnology Co., Ltd. | CS9603-100mL | For tissue mounting |
| Moxa stick | The Third Affiliated Hospital of Zhejiang Chinese Medical University | N/A | For herbs-partitioned moxibustion treatment |
| Pentobarbital sodium | Hangzhou Dawen Biological Co., Ltd. | N/A | For specimen collection |
| Phosphate Buffered Saline (PBS) | Hangzhou Gaosheng Biotechnology Co., Ltd. | P1010-2L | For specimen collection |
| Scissor | Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd. | lsnz | For specimen collection |
| Sectioning blade | Shanghai Saihan Technology Co., Ltd. | LEICA-819-box | For HE staining paraffin section preparation |
| Shaver | Shenzhen Cordes Electrical Appliance Co., Ltd. | N/A | For shaving a rat's hair |
| Sodium Chloride | Chemical Reagent Co., Ltd. | A501218-0001 | For specimen collection |
| Sucrose | Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd. | S11055-500g | For gradient dehydration |
| Thermal insulation container | Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd. | lsbwx8l | For paraffin section dewaxing |
| Tissue embedding cassettes, rectangular in shape, with a breakable lid. | Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd. | 31050102W | For HE staining tissue embedding |
| Wide adhesive tape | Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd. | N/A | For stabilizing rat during the treatment procedure |
References
- Wang, Y., et al. Quality marker discovery of danggui jianzhong decoction for treating primary dysmenorrhoea based on chinmedomics strategy. Phytomedicine. 115, 154724 (2023).
- Wu, L., et al.
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