Summary
The present protocol describes a moxibustion application method for mice, with the benefits of reducing mouse fearfulness, guaranteeing their welfare, and improving efficiency for researchers.
Abstract
The field of moxibustion research is expanding, with a rapid increase in publications in recent years. Moxibustion is a therapy that ignites moxa on the skin of humans, with an increase in peripheral skin temperature and localized redness. During this treatment, the recipient must remain still to prevent scalding and expose intervention sites for easy manipulation; however, maintaining a fixed posture during moxibustion is a big challenge for animals. Thus, manipulating moxibustion in small animals, such as mice, can lead to several difficulties for researchers. In addition, an uncomfortable posture for animals can lead to fear and resistance to moxibustion, increased risk of injury, diminished animal welfare, and less valid research data. An efficient, comfortable moxibustion method is needed to protect animal welfare and minimize the adverse effects on experimental results. However, moxibustion methods are highly variable and often have limited efficacy. More importantly, an uncomfortable moxibustion posture might cause a stress response, such as those observed with anxiety, fear, and anger, which could influence the research data. Therefore, strategies for animal moxibustion that inflict the least harm possible during the intervention are required. This protocol introduces a mouse tethering method for moxibustion intervention, minimizing mouse discomfort and improving study efficiency. Essential strategies for tethering mice and application of moxibustion are highlighted, and the structure of the tethering instrument is described.
Introduction
Moxibustion is an external treatment used in traditional Chinese medicine1. It has been widely applied for centuries as a means of keeping fit and preventing and curing diseases2,3. However, the mechanism of moxibustion remains to be elucidated; thus, this field of research has expanded in recent years as researchers explore its effects and biological mechanisms.
During moxibustion, the recipient must remain still for the best results to be achieved4. Moxibustion treatment typically lasts for 15-30 min5,6. Because of this, manipulation of moxibustion for animals, especially small animals such as mice and rats (favorable for use in animal experiments), has become a big challenge for researchers. Most moxibustion devices are suitable for clinical use and cannot be used for animals, especially for small animals such as mice7,8,9. Besides this, unlike human beings, animals can barely follow orders and remain steady over a while. A fixed and uncomfortable posture for animals can lead to passive emotions, including anxiety, fear, anger, nervousness, and resistance to treatment, increasing the risk of injury, diminishing animal welfare, and resulting in less valid research data10. To solve this problem, in previous studies, anesthetics have been applied for extended periods during animal experiments involving external traditional Chinese medicine therapies, including acupuncture, moxibustion, and massage research11,12,13. However, given that physical conditions change with anesthesia, and in most clinical cases, moxibustion is administered to people in conscious states, anesthesia might impact research data authenticity. Therefore, a method of tethering animals consciously and comfortably become a challenge. To solve these technical problems, specialized devices for mouse moxibustion have been introduced. This protocol provides a tethering method that will assist researchers in performing moxibustion treatment on animals, allowing the animals to remain steady, conscious, and comfortable.
Protocol
Representative Results
Discussion
The moxibustion technique is effective for many disorders, such as fatigue, insomnia, diarrhea, and pain syndrome20,21,22,23,24,25,26. With the benefits of moxibustion treatment for many diseases, the mechanism of this technique has attracted the attention of researchers. The application of …
Disclosures
The authors have nothing to disclose.
Acknowledgements
This work has been supported by grants from the National Natural Science Foundation of China (81704187, 8210152562), Sichuan Science and Technology Program (2019YJ0587, 2018JY0482, 2019YJ0329), Sichuan Academy of Medical Sciences and Sichuan People's Hospital Research Fund (2018ZX05), the Xinglin Scholars Research Foundation of Chengdu University of Traditional Chinese Medicine (QNXZ2019034).
Materials
| Complete Freund's Adjuvant (CFA) | Sigma-Aldrich | SF588102 | |
| Glue gun | Deli Group Co., Ltd. | DL5041 | |
| Laboratory glove | Boci Co., Ltd | 53625130323 | |
| Magnetic metal rectangle (thickness: 0.2 cm) | Sitoo Stationery Co., Ltd | 1007908729 | |
| Moxa stick | Hanyi Airong factory | 5613 | |
| Steel sheet | Rizhan metal materials Co., Ltd | 1108 | |
| Syringe (1000 mL ) | Xinmin Fuda Co., Ltd | 796341 | |
| Timer | Bevoza Co., Ltd | KT003 | |
| Velcro belt | Minnesota Mining and Manufacturing Co., Ltd | 3MSJ3550 | |
| Wooden cube | Chuang Hing Wood Chip Co., Ltd | 2581457A | |
| Wooden splint | Chuang Hing Wood Chip Co., Ltd | 3410968M | |
| Wooden stick | Chuang Hing Wood Chip Co., Ltd | 2785476M | |
| Wooden strip | Chuang Hing Wood Chip Co., Ltd | 2374652S |
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