Du-Moxibustion in a Mouse Model of Ankylosing Spondylitis

Summary

This paper describes in detail the operation procedures and precautions to be taken during Du-moxibustion in the treatment of ankylosing spondylitis in experimental mice.

Abstract

Ankylosing spondylitis (AS) is a progressively worsening and disabling form of arthritis that primarily affects the axial skeleton. This disease mainly involves the spine and the sacroiliac joint. Fusion of the spine and the sacroiliac joint may occur in the later stage of the disease, resulting in spinal stiffness and kyphosis, as well as difficulty in walking, which seriously affects the quality of work and daily living activities and imposes a heavy burden on the patient, the family, and society. Increasing attention has been paid to non-pharmacotherapy as an alternative therapy for AS. Moxibustion is an ancient therapeutic technique used in Traditional Chinese Medicine (TCM). Du-moxibustion therapy, a unique and innovative external treatment developed on the basis of ordinary moxibustion, has a definite therapeutic effect on AS. Du-moxibustion skillfully combines the compatible techniques of TCM to integrate meridians, acupoints, Chinese herbal medicine, and moxibustion. This paper describes the operation procedures and precautions to be taken during Du-moxibustion in experimental mice in detail to provide an experimental basis for the study of the mechanism of Du-moxibustion in the treatment of AS.

Introduction

Ankylosing spondylitis (AS) is a progressively worsening and disabling form of arthritis that primarily affects the axial skeleton. According to the latest epidemiological survey, ~0.01%-1.8% of people in the world suffer from this condition¹, most of whom are young males (male to female ratio is ~2-3:1)². The incidence of AS in the Chinese population is 0.2%-0.4%. This disease mainly involves the spine and the sacroiliac joint. Fusion of the spine and the sacroiliac joint may occur in the later stages of the disease, resulting in spinal stiffness and kyphosis, as well as difficulty in walking, which seriously affects th....

Protocol

All animal experiments were approved by the Experimental Animal Welfare Ethics Review Committee of Shandong University of Traditional Chinese Medicine (No. SDUTCM20230831301).

1. Animal preparation

1. Determine the experimental groups and the number of animals per group in advance.
2. Choose male BALb/c mice, house them at a constant temperature of 22 ± 1 °C, constant humidity of 60 ± 5%, and 12 h diurnal cycle light environment.

Discussion

AS is clinically manifested as back, lumbar, and joint swelling pain, and spinal deformity and joint rigidity may occur in severe cases²⁶. Early diagnosis and treatment can improve the quality of life, reduce disability rates, and improve the prognoses of patients with AS. According to traditional Chinese medicine (TCM), AS is believed to be caused by a deficiency of Yang in the kidney and the Du meridian, as well as an accumulation of cold evil internally. TCM holds the view that moxibustion, bas.......

Materials

Name	Company	Catalog Number	Comments
10% EDTA	Shanghai Macklin Biochemical Co., Ltd.,Shanghai, China
alcohol pads	HYNAUT, Qingdao Hainuo Biological Engineering Co., LTD, Qingdao, China
anesthesia machine	Medical Supplies & Services INT. LTD, Keighley, UK
centrifuge tubes	Axygen, Corning, NewYork, UAS
complete Frech's adjuvant (CFA)	aladdin,Shanghai, China	F393378
cotton ball	Henan RUIKE MEDICAL Equipment Co., Ltd.,Xinxiang, China
cotton gauze	Henan RUIKE MEDICAL Equipment Co., Ltd.,Xinxiang, China
cutting board	self-preparation
decorin from bovine articular cartilage	Sigma-Aldrich, MO, USA	D8428
depilatory cream	Veet, Reckitt Benckiser (China) Investment Co. LTD, Shanghai, China
electronic scale	Shanghai Yajin Electronic Technology Co., Ltd.,Shanghai, China
Eppendorf tube	Axygen, Corning, NewYork, UAS
eye lubricant	Beijing Shuangji Pharmaceutical Co., LTD., Beijing, China
ginger	self-preparation
GraphPad Prism 7 software	GraphPad Software,Boston, USA
hair clipper	Super human Group CO LTD, Jinhua, China
heating pads	Shenzhen Leshuo Tech Co., Ltd.,Shenzhen, China
incomplete Freund’s adjuvant (IFA)	aladdin,Shanghai, China	F393371
injection syringe	Shandong Xinhua Ande Medical Supplies Co., LTD, Zibo, China
isoflurane	Shenzhen Rayward Life Technology Co., LTD, Shenzhen, China	R510-22-10
joss stick	Shijiazhuang Lidu Fragrant Industry Co., LTD.,Shijiazhuang, China
juicer	Braun (Shanghai) Co., Ltd.,Shanghai, China
knife	self-preparation
lighter	Zhejiang tiger-lighter Co. LTD
mortar	Luoyang Yinai Ceramic Technology Co., LTD.,Luoyang, China
Mouse IL-17 ELISA Kit	absin, Shanghai, China	abs520009-96T
Mouse TNF-α ELISA Kit	Wuhan Sanying, Wuhan, China	KE10002
mulberry paper	Yishui County Mulinsang paper Co., LTD, Linyi, China
OFT	Xinruan,Shanghai, China	XR-XZ301
paper cup	self-preparation
pipettes	OXFORD BIO INSTRUMENTS INC.,Oxford, UK
refined moxa velvet	self-preparation
rotarod	Xinruan,Shanghai, China	XR-6C
scientz-48L cryogenic high throughput tissue grinder	Ningbo Xinzhi Biotechnology Co., LTD
scissors	Shandong Jiaren Medical Supplies Co., Ltd., Zibo, China
semi-automatic paraffin sectioning machine	Leica Camera AG, Watznach, Germany
SPSS 25.0 software	International Business Machines Corporation, NewYork, UAS
TCM powder	self-preparation
tips	Biosharp, Labgic, Beijing, China
writing brush	Yishui County Mulinsang paper Co., LTD, Linyi, China