This work was supported by&#160;the Zhejiang Provincial Natural Science&#160;Foundation, Exploring Program Q&#160;(LQ20H270012), Zhejiang Traditional Chinese&#160;Medicine Technology Plan (NO.2020ZB124), the&#160;Research Project of Zhejiang Chinese Medical&#160;University (NO.2021JKZKTS056B), and the&#160;Hospital-Level Project of the Third Affiliated&#160;Hospital of Zhejiang Chinese Medical&#160;University (NO.ZS22ZA01). We are grateful to&#160;the Experimental Animal Center of Zhejiang&#160;Chinese Medical University and the Key&#160;Laboratory of Acupuncture and Neurology of&#160;Zhejiang Province for providing us with the&#160;experimental conditions.

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...

Herbal Moxibustion on Shenque Acupoint for Alleviating Primary Dysmenorrhea

<ol>
	<li>Wang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quality+marker+discovery+of+danggui+jianzhong+decoction+for+treating+primary+dysmenorrhoea+based+on+chinmedomics+strategy.">Quality marker discovery of danggui jianzhong decoction for treating primary dysmenorrhoea based on chinmedomics strategy.</a> Phytomedicine. 115, 154724 (2023).</li><li>Wu, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unraveling+the+anti-primary+dysmenorrhea+mechanism+of+ainsliaea+fragrans+champ.+Extract+by+the+integrative+approach+of+network+pharmacology+and+experimental+verification.">Unraveling the anti-primary dysmenorrhea mechanism of ainsliaea fragrans champ. Extract by the integrative approach of network pharmacology and experimental verification.</a> Phytomedicine. 123, 155213 (2024).</li><li>Proctor, M., Farquhar, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnosis+and+management+of+dysmenorrhoea.">Diagnosis and management of dysmenorrhoea.</a> BMJ. 332 (7550), 1134-1138 (2006).</li><li>Unsal, A., Ayranci, U., Tozun, M., Arslan, G., Calik, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevalence+of+dysmenorrhea+and+its+effect+on+quality+of+life+among+a+group+of+female+university+students.">Prevalence of dysmenorrhea and its effect on quality of life among a group of female university students.</a> Ups J Med Sci. 115 (2), 138-145 (2010).</li><li>Jamieson, D. J., Steege, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+prevalence+of+dysmenorrhea,+dyspareunia,+pelvic+pain,+and+irritable+bowel+syndrome+in+primary+care+practices.">The prevalence of dysmenorrhea, dyspareunia, pelvic pain, and irritable bowel syndrome in primary care practices.</a> Obstet Gynecol. 87 (1), 55-58 (1996).</li><li>Andersch, B., Milsom, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+epidemiologic+study+of+young+women+with+dysmenorrhea.">An epidemiologic study of young women with dysmenorrhea.</a> Am J Obstet Gynecol. 144 (6), 655-660 (1982).</li><li>Watanabe, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preconception+dysmenorrhea+as+a+risk+factor+for+psychological+distress+in+pregnancy:+The+japan+environment+and+children's+study.">Preconception dysmenorrhea as a risk factor for psychological distress in pregnancy: The japan environment and children's study.</a> J Affect Disord. 245, 475-483 (2019).</li><li>Bajalan, Z., Moafi, F., Moradibaglooei, M., Alimoradi, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mental+health+and+primary+dysmenorrhea:+A+systematic+review.">Mental health and primary dysmenorrhea: A systematic review.</a> J Psychosom Obstet Gynaecol. 40 (3), 185-194 (2019).</li><li>Dong, Y., Li, M. J., Hong, Y. Z., Li, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insight+into+dysmenorrhea+research+from+1992+to+2022:+A+bibliometric+analysis.">Insight into dysmenorrhea research from 1992 to 2022: A bibliometric analysis.</a> J Pain Res. 16, 3591-3611 (2023).</li><li>Iacovides, S., Avidon, I., Bentley, A., Baker, F. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduced+quality+of+life+when+experiencing+menstrual+pain+in+women+with+primary+dysmenorrhea.">Reduced quality of life when experiencing menstrual pain in women with primary dysmenorrhea.</a> Acta Obstet Gynecol Scand. 93 (2), 213-217 (2014).</li><li>Wang, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+repeated+menstrual+pain+on+empathic+neural+responses+in+women+with+primary+dysmenorrhea+across+the+menstrual+cycle.">Effects of repeated menstrual pain on empathic neural responses in women with primary dysmenorrhea across the menstrual cycle.</a> Hum Brain Mapp. 42 (2), 345-356 (2021).</li><li>Harel, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyclooxygenase-2+specific+inhibitors+in+the+treatment+of+dysmenorrhea.">Cyclooxygenase-2 specific inhibitors in the treatment of dysmenorrhea.</a> J Pediatr Adolesc Gynecol. 17 (2), 75-79 (2004).</li><li>Zahradnik, H. P., Hanjalic-Beck, A., Groth, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonsteroidal+anti-inflammatory+drugs+and+hormonal+contraceptives+for+pain+relief+from+dysmenorrhea:+A+review.">Nonsteroidal anti-inflammatory drugs and hormonal contraceptives for pain relief from dysmenorrhea: A review.</a> Contraception. 81 (3), 185-196 (2010).</li><li>Strowitzki, T., Kirsch, B., Elliesen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+of+ethinylestradiol+20+&#181;g/drospirenone+3+mg+in+a+flexible+extended+regimen+in+women+with+moderate-to-severe+primary+dysmenorrhoea:+An+open-label,+multicentre,+randomised,+controlled+study.">Efficacy of ethinylestradiol 20 &#181;g/drospirenone 3 mg in a flexible extended regimen in women with moderate-to-severe primary dysmenorrhoea: An open-label, multicentre, randomised, controlled study.</a> J Fam Plann Reprod Health Care. 38 (2), 94-101 (2012).</li><li>Liu, Y., Sun, J., Wang, X., Shi, L., Yan, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+herb-partitioned+moxibustion+for+primary+dysmenorrhea:+A+randomized+clinical+trial.">Effect of herb-partitioned moxibustion for primary dysmenorrhea: A randomized clinical trial.</a> J Tradit Chin Med. 39 (2), 237-245 (2019).</li><li>Li, S. -. J., Pan, Y. -. H., Zhao, Q. T., Gao, S. -. Z., Ma, Y. -. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+mast+cell+function+on+umbilical+therapy+for+uterine+transient+receptor+potential+vanillin+pathway+in+rats+with+dysmenorrhea+of+cold+congelation+and+blood+stasis+syndrome.">Effects of mast cell function on umbilical therapy for uterine transient receptor potential vanillin pathway in rats with dysmenorrhea of cold congelation and blood stasis syndrome.</a> China Journal of Traditional Chinese Medicine and Pharmacy. 33 (10), 4646-4649 (2018).</li><li>Ma, Y. X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Research+of+herb-partitioned+moxibustion+for+primary+dysmenorrhea+patients+based+on+the+lc-ms+metabonomics.">Research of herb-partitioned moxibustion for primary dysmenorrhea patients based on the lc-ms metabonomics.</a> Evid Based Complement Alternat Med. 2015, 621490 (2015).</li><li>Tian, T., Li, L., Hong, H., Ying, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+understanding+and+treatment+experience+of+dysmenorrhea+by+doctors+in+different+dynasties.">The understanding and treatment experience of dysmenorrhea by doctors in different dynasties.</a> Henan Journal of Traditional Chinese Medicine. 37 (1), 50-52 (2017).</li><li>Jiaqi, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TCM+pattern+distribution+characteristics+of+primary+dysmenorrhea.">TCM pattern distribution characteristics of primary dysmenorrhea.</a> Shenzhen Journal of Integrated Traditional Chinese and and Western Medicine. 30 (13), 47-49 (2020).</li><li>Fang, C., Junhong, C., Wei, D., Namei, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exploration+of+TCM+Syndrome+Distribution+Characteristics+of+Primary+Dysmenorrhea.">Exploration of TCM Syndrome Distribution Characteristics of Primary Dysmenorrhea.</a> Acta Chinese Medicine. 23 (08), 1194-1196 (2013).</li><li>Tian, T., Li, L., Hong, H., Ying, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recognition+of+dysmenorrhea+by+medical+practitioners+through+the+ages+and+their+experience+in+diagnosis+and+treatment+of+dysmenorrhea.">Recognition of dysmenorrhea by medical practitioners through the ages and their experience in diagnosis and treatment of dysmenorrhea.</a> Henan Traditional Chinese Medicine. 37 (01), 50-52 (2017).</li><li>Ren Qing, W. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progress+in+treatment+of+primary+dysmenorrhea+with+traditional+Chinese+and+Western+medicine.">Progress in treatment of primary dysmenorrhea with traditional Chinese and Western medicine.</a> Journal of Practical Traditional Chinese Internal Medicine. 34 (07), 4-7 (2020).</li><li>Siyi, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characteristics+of+acupoints+selection+of+moxibustion+for+primary+dysmenorrhea+based+on+data+mining+technology.">Characteristics of acupoints selection of moxibustion for primary dysmenorrhea based on data mining technology.</a> Zhongguo Zhen Jiu. 35 (08), 845-849 (2015).</li><li>Yang, Y. E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Argyi+research+and+application+in+ancient+and+modern+plague.">Argyi research and application in ancient and modern plague.</a> Lishizhen Medicine and Materia Medica Research. 31 (02), 438-441 (2020).</li><li>Yan, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+and+mechanism+of+warm+moxibustion+at+shenque+acupoint+in+treating+primary+dysmenorrhea+in+college+students.">Effect and mechanism of warm moxibustion at shenque acupoint in treating primary dysmenorrhea in college students.</a> Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology. , (2024).</li><li>Yang, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Effect+of+Acupuncture+to+SP6+on+Skin+Temperature+Changes+of+SP6+and+SP10:+An+Observation+of+“Deqi”.">The Effect of Acupuncture to SP6 on Skin Temperature Changes of SP6 and SP10: An Observation of “Deqi”.</a> Evidence-Based Complementary and Alternative Medicine. 2014 (1), 1741-4288 (2014).</li><li>Al, X. Y. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+and+assessment+method+of+blood+stasis+due+to+cold+coagulation+dysmenorrhea+rat+model+research.">Preparation and assessment method of blood stasis due to cold coagulation dysmenorrhea rat model research.</a> Acta Chinese Medicine and Pharmacology. 41 (04), 25-30 (2013).</li><li>Du Dongqing, Y. C., Ma, Y., Liu, X., Gao, S., Wang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+study+of+herb-partitioned+moxibustion+on+navel+for+primary+dysmenorrhea.">Clinical study of herb-partitioned moxibustion on navel for primary dysmenorrhea.</a> Shanghai Journal of Acupuncture and Moxibustion. 30 (08), 514-516 (2011).</li><li>Wang Haijun, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Location+of+shenque+acupoint+in+rats.">Location of shenque acupoint in rats.</a> Acupuncture Research. 5, 1 (2007).</li><li>Wei, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+moxibustion+at+&amp;#34;oppositely-located+points&amp;#34;+on+neurogenic+bladder+afterspinal+cord+injury+and+endoplasmic+reticulum+stress+pathway+in+rats.">Effect of moxibustion at &amp;#34;oppositely-located points&amp;#34; on neurogenic bladder afterspinal cord injury and endoplasmic reticulum stress pathway in rats.</a> Zhongguo Zhen Jiu. 42 (04), 413-418 (2022).</li><li>Schmauss, C., Yaksh, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+studies+on+spinal+opiate+receptor+systems+mediating+antinociception.+II.+Pharmacological+profiles+suggesting+a+differential+association+of+mu,+delta+and+kappa+receptors+with+visceral+chemical+and+cutaneous+thermal+stimuli+in+the+rat.">In vivo studies on spinal opiate receptor systems mediating antinociception. II. Pharmacological profiles suggesting a differential association of mu, delta and kappa receptors with visceral chemical and cutaneous thermal stimuli in the rat.</a> J Pharmacol Exp Ther. 228 (1), 1-12 (1984).</li><li>Zhao, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Herb-partitioned+moxibustion+improves+crohn's+disease-associated+intestinal+fibrosis+by+suppressing+the+RhoA/ROCK1/MLC+pathway.">Herb-partitioned moxibustion improves crohn's disease-associated intestinal fibrosis by suppressing the RhoA/ROCK1/MLC pathway.</a> Evid Based Complement Alternat Med. 2021, 2247953 (2021).</li><li>Yan, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+herb-partitioned+moxibustion+for+ulcerative+colitis:+A+protocol+for+systematic+review+and+meta-analysis.">Effects of herb-partitioned moxibustion for ulcerative colitis: A protocol for systematic review and meta-analysis.</a> Medicine (Baltimore). 99 (31), e21319 (2020).</li><li>Dai, X. Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Moxibustion+strategies+for+mice.">Moxibustion strategies for mice.</a> J Vis Exp. 179, 63119 (2022).</li></ol>

The authors have no conflicts of interest to declare.

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 &#176;C. In the previous pre-experiment, it was found that when the temperature was &#8722;25 &#176;C, the rats would have...

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 and thighs are the characteristic symptoms of polycystic PDM. Due to differing definitions of the condition and a lack of standardized ways to evaluate the intensity of menstrual pain, the estimated prevalence of dysmenorrhea in women varies from 45%-95%3,4,5. About 10%-25% of women who are of reproductive age are thought to suffer from really severe PDM6. The prevalence of PDM prevalence is highest in the 20- to 24-year-old age group and decreases progressively thereafter2. PDM is characterized by periodic lower abdominal pain, which is frequently accompanied by other systemic symptoms such as low back pain, nausea, vomiting, headaches, anorexia, diarrhea7, and even mental symptoms like depression and anxiety8,9. During menstruation, women with dysmenorrhea experience a decline in overall quality of life4,8,10.
Furthermore, studies have found that repeated menstrual pain may affect the function of the central nervous system, leading to altered perception of pain in both painful or pain-free stages11. Due to the etiology of PDM being based on prostaglandins (PGs), the most common drug treatment for PDM is nonsteroidal anti-inflammatory drugs (NSAIDs)12,13, followed by oral contraceptives14. In addition to the side effects of drugs, 10%-20% of people do not respond to NSAIDs and contraceptives3. The majority of current research is concerned with the effects of dysmenorrhea on brain anatomy and physiology, with little attention paid to therapeutic approaches. Consequently, there is an urgent need to find a treatment strategy that can be widely applied in clinical practice, with established efficacy, and readily accepted by patients.
Herbs-partitioned moxibustion (HPM) is a characteristic external therapy in traditional Chinese medicine (TCM), which is widely used to treat diseases in China. Its benefits include minimal cost, no side effects, good long-term effects, easy operation, and easy acceptance by patients. It has been found that the effect of HPM on the navel on PDM is significant15, which may be closely related to regulating the cold and heat deficiency and excess of the uterus as well as mast cells and related receptors16. The research team has proved that HPM can immediately relieve menstrual pain17.
Zhang Zhong-jing initially recorded the concept of dysmenorrhea in the Women &#39;s Miscellaneous Diseases and Treatment of the Golden Chamber, writing that the menstrual flow is unfavorable under the belt, and the abdomen is full of pain18. According to its main symptoms, dysmenorrhea can be attributed to the category of menstrual pain and menstrual abdominal pain in traditional Chinese medicine. The syndromes of dysmenorrhea can be divided into five: qi and blood deficiency; cold coagulation and blood stasis; damp heat accumulation; qi stagnation and blood stasis; and liver and kidney deficiency16. According to relevant studies, the syndrome of cold coagulation and blood stasis is the most common syndrome of PDM19,20. TCM holds that PDM and cold pathogens are intimately connected21. During or around menstruation, there is an excessive flow of blood in Xuehai, leading to a quick depletion of qi-blood due to its energetic nature. Consequently, women are vulnerable to disease-causing causes due to a lack of sufficient healthy energy and the rapid fluctuations of qi and blood in the uterus and Chong and Ren meridians22. Women who have well-developed physiques should take precautions to avoid consuming raw or cold food and to prevent catching a cold during menstruation. Failure to do so may result in the intrusion of cold pathogens into the uterus, leading to the coagulation and stagnation of blood in the uterus and the Chong and Ren meridians. Consequently, individuals experience pain as a result of the stagnation of blood flow. Both the intrusion of cold pathogens from the outside and the presence of yin cold within the body can cause blockage and clotting of the meridians.
The location of dysmenorrhea is in the Chong and Ren meridians, and the onset is related to changes in qi and blood. In TCM theory, moxibustion works by stimulating Yang Qi, warming the meridians, dispersing cold, and increasing blood flow to relieve pain. According to Jinfu of Lingshu23, if the pulse and blood clot are in the middle, and there is congealed blood and cold blood, then it is appropriate for moxibustion; it is a typical external dysmenorrhea treatment. In a previously published work24 artemisia argyi leaves are described as bitter and pungent, capable of restoring the nearly exhausted original Yang, connecting the twelve meridians, running through the three Yin meridians, regulating Qi and blood, dispelling cold and dampness, and warming the uterus, moxibustion, with the help of corresponding acupoints, can expel pathogenic factors externally, regulate qi and blood internally, and thereby play a role in regulating the meridians, activating blood circulation, and stopping pain. The navel, also known as the Shenque (CV8), belongs to the Ren meridian, connecting the Du, Chong, and Dai meridians. According to TCM theory, Shenque is believed to be related to the five zang and six fu25. It is regarded as a congenital foundation and the pivot of the yin-yang balance. Anatomical, the PDM is in close proximity to the pelvis and is closely connected to the uterus. The PDM disease specifically affects the uterus, and its pathogenesis involves an imbalance of qi and blood as well as an imbalance of the viscera. By stimulating the Shenque point, it is possible to directly influence the qi and blood condition of the uterus and effectively treat menstrual cramps. Based on the aforementioned theory, we selected the stimulation of the Shenque acupoint located in the abdominal region for HPM therapy, with the aim of ameliorating dysmenorrhea symptoms and alleviating pain.
To mimic the intervention of TCM on PDM as closely as possible with cold coagulation and blood stasis, we replicated the HPM technique on the navel in female rats. It was discovered that HPM on the navel has a clear analgesic effect and can lessen the writhing reaction of dysmenorrhea rats. In order to investigate the mechanism of HPM on the navel to improve PDM and promote it, we have examined a technique of fixing rats and performing HPM at the Shenque acupoint in the belly.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.9% physiological saline</td>
			<td>Hangzhou Gaosheng Biotechnology Co., Ltd.</td>
			<td>SLYSZ1A</td>
			<td>For injecting the control group&#160;</td>
		</tr>
		<tr>
			<td>1 mL disposable sterile syringe</td>
			<td>Changzhou Yuekang Medical Equipment Co., Ltd.</td>
			<td>3000-0515-14</td>
			<td>For drug injection</td>
		</tr>
		<tr>
			<td>3 mL (extended) Pasteur pipette</td>
			<td>Beijing Lanjieke Technology Co., Ltd.</td>
			<td>YCKJ-HC-007245</td>
			<td>For mounting</td>
		</tr>
		<tr>
			<td>75% disinfection alcohol</td>
			<td>Hangzhou OUTOP Biotechnology Co., Ltd.</td>
			<td>N/A</td>
			<td>For sterilizing the treatment site</td>
		</tr>
		<tr>
			<td>Adhesive-coated glass slide</td>
			<td>Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd.</td>
			<td>188105W</td>
			<td>For HE staining</td>
		</tr>
		<tr>
			<td>Biochemical incubator</td>
			<td>Shanghai Boxun Industrial Co., Ltd. Medical Equipment Factory</td>
			<td>N/A</td>
			<td>For tissue dewaxing in HE staining</td>
		</tr>
		<tr>
			<td>Cloth material</td>
			<td>In-house preparation</td>
			<td>N/A</td>
			<td>Pouch to hold rat</td>
		</tr>
		<tr>
			<td>Digital manager slice scanning system</td>
			<td>Hamamatsu</td>
			<td>C13210-01</td>
			<td>For HE slice scanning</td>
		</tr>
		<tr>
			<td>Disposable latex gloves</td>
			<td>Guangzhou Lige Technology Co., Ltd.</td>
			<td>LG10-102-1I</td>
			<td>For use during operation</td>
		</tr>
		<tr>
			<td>Estradiol benzoate</td>
			<td>Hefei Xinkexin Animal Pharmaceutical Co., Ltd.</td>
			<td>N/A</td>
			<td>For establishing dysmenorrhea model</td>
		</tr>
		<tr>
			<td>Excelsior AS Fully automatic dehydrator</td>
			<td>Thermo Scientific</td>
			<td>Excelsior AS</td>
			<td>For HE staining tissue dehydration</td>
		</tr>
		<tr>
			<td>Female Wistar rats</td>
			<td>Laboratory Animal Center of Zhejiang Chinese Medicine University</td>
			<td>N/A</td>
			<td>220&#8211;250 g</td>
		</tr>
		<tr>
			<td>Flour</td>
			<td>In-house preparation</td>
			<td>N/A</td>
			<td>For herbs-partitioned moxibustion treatment</td>
		</tr>
		<tr>
			<td>Forcep</td>
			<td>Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd.</td>
			<td>lsnz</td>
			<td>For specimen collection</td>
		</tr>
		<tr>
			<td>Formaldehyde&#160;</td>
			<td>Shanghai Lingfeng Chemical Reagent Co., Ltd.</td>
			<td>441244-1KG</td>
			<td>For tissue fixation</td>
		</tr>
		<tr>
			<td>Gemini AS slide stainer</td>
			<td>Thermo Scientific</td>
			<td>Gemini AS&#160;</td>
			<td>Staining machine</td>
		</tr>
		<tr>
			<td>Hemostat</td>
			<td>Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd.</td>
			<td>lsnz</td>
			<td>For specimen collection</td>
		</tr>
		<tr>
			<td>HistoCore</td>
			<td>Leica ( Arcadia H )</td>
			<td>14039357258</td>
			<td>For HE staining tissue embedding</td>
		</tr>
		<tr>
			<td>Medical cotton ball sterile</td>
			<td>The Third Affiliated Hospital of Zhejiang Chinese Medical University</td>
			<td>N/A</td>
			<td>For sterilize the tratment site</td>
		</tr>
		<tr>
			<td>Methylene blue staining solution (0.1%)</td>
			<td>Beijing Honghu United Chemical Products Co., Ltd.</td>
			<td>G1300</td>
			<td>For screening estrous cycle</td>
		</tr>
		<tr>
			<td>Microdissection Shears</td>
			<td>Hangzhou Gaosheng Biotechnology Co., Ltd.</td>
			<td>XWJ12A</td>
			<td>For specimen collection</td>
		</tr>
		<tr>
			<td>Microscope cover glass (24 mm x 50 mm)</td>
			<td>Hangzhou Zhengbo Biotechnology Co., Ltd.</td>
			<td>10212450C</td>
			<td>For HE staining</td>
		</tr>
		<tr>
			<td>Microtome</td>
			<td>Leica Microsystems</td>
			<td>RM2255</td>
			<td>For tissue sectioning</td>
		</tr>
		<tr>
			<td>Midea freezer</td>
			<td>Hefei Midea Refrigerator Co., Ltd.</td>
			<td>N/A</td>
			<td>For establishing dysmenorrhea model with cold coagulation and blood stasis</td>
		</tr>
		<tr>
			<td>Molar forceps</td>
			<td>Hangzhou Zhengbo Biotechnology Co., Ltd.</td>
			<td>YGJD-11</td>
			<td>For specimen collection</td>
		</tr>
		<tr>
			<td>Mounting medium</td>
			<td>Hangzhou Gaosheng Biotechnology Co., Ltd.</td>
			<td>CS9603-100mL</td>
			<td>For tissue mounting</td>
		</tr>
		<tr>
			<td>Moxa stick</td>
			<td>The Third Affiliated Hospital of Zhejiang Chinese Medical University</td>
			<td>N/A</td>
			<td>For herbs-partitioned moxibustion treatment</td>
		</tr>
		<tr>
			<td>Pentobarbital sodium</td>
			<td>Hangzhou Dawen Biological Co., Ltd.</td>
			<td>N/A</td>
			<td>For specimen collection</td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline (PBS)</td>
			<td>Hangzhou Gaosheng Biotechnology Co., Ltd.</td>
			<td>P1010-2L</td>
			<td>For specimen collection</td>
		</tr>
		<tr>
			<td>Scissor&#160;</td>
			<td>Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd.</td>
			<td>lsnz</td>
			<td>For specimen collection</td>
		</tr>
		<tr>
			<td>Sectioning blade</td>
			<td>Shanghai Saihan Technology Co., Ltd.</td>
			<td>LEICA-819-box</td>
			<td>For HE staining paraffin section preparation</td>
		</tr>
		<tr>
			<td>Shaver</td>
			<td>Shenzhen Cordes Electrical Appliance Co., Ltd.</td>
			<td>N/A</td>
			<td>For shaving a rat&#39;s hair</td>
		</tr>
		<tr>
			<td>Sodium Chloride</td>
			<td>Chemical Reagent Co., Ltd.</td>
			<td>A501218-0001</td>
			<td>For specimen collection</td>
		</tr>
		<tr>
			<td>Sucrose</td>
			<td>Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd.</td>
			<td>S11055-500g</td>
			<td>For gradient dehydration</td>
		</tr>
		<tr>
			<td>Thermal insulation container</td>
			<td>Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd.</td>
			<td>lsbwx8l</td>
			<td>For paraffin section dewaxing</td>
		</tr>
		<tr>
			<td>Tissue embedding cassettes, rectangular in shape, with a breakable lid.</td>
			<td>Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd.</td>
			<td>31050102W</td>
			<td>For HE staining tissue embedding</td>
		</tr>
		<tr>
			<td>Wide adhesive tape</td>
			<td>Zhejiang Tuzhi Pharmaceutical Technology Co., Ltd.</td>
			<td>N/A</td>
			<td>For stabilizing rat during the treatment procedure</td>
		</tr>
	</tbody>
</table>

herbs-partitioned moxibustion on the navel in a rat model of primary dysmenorrhea with cold coagulation and blood stasis

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.

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

Research on the traditional Chinese medicine anesthetic method of primary dysmenorrhea. We want to check if herbs-partitioned moxibustion on the navel can relieve the pain of dysmenorrhea rats. Keeping the rats quiet during the experiment is an experimental challenge that we encountered.We intend to study the reasons behind the chronic pain in dysmenorrhea and would like to explore the anesthetic mechanism of acupuncture and moxibustion. To begin, obtain the adaptive diet-fed female rat that was maintained in a 12-hour light-dark cycle. Insert the pipette gun with 60 microliters of 0.9%saline into the vaginal orifice of the rat.Pipette up and down three to four times. Suck out the liquid and drop it onto a glass slide. To fix the dried content, add 60 microliters of 95%ethanol.After eight minutes, rinse and dry the content. Add 0.1%methylene blue staining solution and incubate away from light for 10 minutes. Afterward, rinse and dry the content.Observe the slide under the microscope and obtain images to screen the rat's estrus cycle. To establish primary dysmenorrhea, inject the rat with estradiol benzoate subcutaneously in the thigh for 10 days. Place the female rat in a refrigerator at 20 degrees Celsius for four hours on the first to fifth day of modeling.To begin, establish a primary dysmenorrhea rat model using estradiol benzoate injections. Obtain the moxa cones for herbs-partitioned moxibustion or HPM navel therapy and rub the moxa floss in the hand into a spindle shape. Prepare a dough bowl with a hole in the middle.To prepare the rat pouch, sew a cut cloth together diagonally. Cut out a small opening at the top for breathing and a hole on one side of the pouch for moxibustion. Then sew a pair of cloth ropes of about 10 centimeters in length and 1 centimeter in width at the top and bottom ends of the bag respectively for tethering.Next, locate the shenque acupoint on the midline of the rat's abdomen. Remove the hair around the shenque acupoint. Place the rat in the rat pouch, keeping the rat's abdomen up, and expose the lower limbs and tail.Fix the upper and lower parts of the rat's body and the tail on the operating table with tape. Disinfect the shenque and its surrounding skin with 75%ethanol. Place the bowl on the abdomen, aiming its hole at the shenque acupoint, and put 0.3 grams of the traditional Chinese medicine powder on the acupoint.Then put the moxa cone on the traditional Chinese medicine powder. Light the moxa cone and treat the rat for 30 minutes. To begin, offer the herbs-partitioned moxibustion navel therapy to a rat with primary dysmenorrhea.Wipe the experimental box with 75%ethanol before each trial. Place the rat in the cage after intraperitoneal injection of oxytocin or 0.9%saline. Record the writhing behavior of the rat for 30 minutes with a video camera.After anesthetizing, determine the weight of the treated rat. Incise the skin and muscle from the middle of the abdomen to expose the heart. Insert a profusion needle into the heart from the apex position, fix the profusion needle with hemostatic forceps, and make a small cut in the right auricle.Perfuse 0.9%saline through the left ventricle and descending aorta until the liver and eyes of the rat become lighter and white in color. Finally, using scissors, cut out the uterus and weigh it accurately to calculate the uterine index. The model group of PDM, with cold coagulation and blood stasis, had a significant increase in writhing score compared to the control group, and the treatment group showed a significant decrease in writhing score.The model group had a significantly reduced writhing latency compared to the control group, and the treatment group showed an increase in writhing latency compared to the model group. The uterine index increased in the model group compared to the control group and decreased in the treatment group compared to the model group.

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&#39;s writhing behavior, determine whether the PDM rat with cold coagulation and blood stasis was successfully modeled, and evaluate the rat&#39;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...

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.

Primary dysmenorrhea (PDM) refers to the occurrence of spasmodic colicky pain in the lower abdomen without any obvious pelvic pathology, often accompanied ...

herbs-partitioned-moxibustion-on-navel-rat-model-primary-dysmenorrhea

Research

JoVE Journal

Medicine

Establishment of Primary Dysmenorrhea Rat Model with Cold Coagulation and Blood Stasis for Herbs-Partitioned Moxibustion Treatment

Herbs-Partitioned Moxibustion Navel Therapy for Treating Primary Dysmenorrhea in Rats

Behavioral Test and Uterine Index Determination to Assess the Efficacy of Herbs-Partitioned Moxibustion in Treating Primary Dysmenorrhea

293 Views.  The Third Clinical Medical College of Zhejiang Chinese Medical University.  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.

Herbal Moxibustion on Shenque Acupoint for Alleviating Primary Dysmenorrhea

Summary