Herbs-Partitioned Moxibustion on the Navel in a Rat Model of Primary Dysmenorrhea with Cold Coagulation and Blood Stasis

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 flow^3,4, 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.......

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.......

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