Using Herbal-Cake-Separated Moxibustion for the Treatment of Rats with Chronic Renal Failure

Summary

The moxibustion experimental platform can fix the rats without restriction, and the rats can undergo multi-acupoint moxibustion in the awake state, ensuring the welfare of the rats, protecting the safety of the operators, ensuring therapeutic efficacy, and improving the efficiency of the experiment.

Abstract

In the process of moxibustion in clinical practice, subjects need to be in a stable mood and comfortable posture to avoid problems such as moxa ash falling, scalding skin, and poor curative effect. Such problems also exist in the rat moxibustion experiment. To simulate clinical practice, it is necessary to introduce an experimental instrument in animal experiments, that is, a moxibustion device with fixed rats and moxibustion treatment synchronization, which can make experimental rats receive moxibustion treatment quietly and comfortably under non-anesthesia.

Our research group designed a rat moxibustion experimental platform. The device was framed by a wooden board with a supporting base plate, multiple fixed components, and partitioned components. The device can achieve the operation mode of moxibustion in rats without binding, avoiding anesthesia and scalding and simultaneously exposing multiple acupoints on the back. This operation can avoid physical and mental injury to rats and operators, which improves the research efficiency and further promotes the development and research of moxibustion animal experiments.

The device has a simple structure, is easy to operate and popularize, is comprehensively and innovatively designed, reusable, and is suitable for rat experiments mainly based on moxibustion. This article mainly introduces the structure of the experimental platform device for rat moxibustion, the basic procedure of herbal-cake-separated-moxibustion in experimental rats using the device and describes the establishment of a rat model of chronic renal failure (CRF) and representative experimental results.

Introduction

Moxibustion appeared with the application of fire. Ancient doctors accidentally found that some parts of the fire-burning body could relieve the symptoms of discomfort. After long-term practice, "Artemisiae Argyi Folium" was gradually selected as the main moxibustion material¹. Moxibustion is one of the important components of traditional Chinese medicine, and when juxtaposed with traditional Chinese medicine decoction and acupoint acupuncture, these three major traditional Chinese medicine therapies are widely used in many diseases.

Chinese doctors often used moxibustion to treat diseases as early as the per....

Protocol

The plan follows the Animal Care Guidelines of the Yunnan University of Chinese Medicine and has been approved by the Animal Ethics Committee of Yunnan University of Chinese Medicine. The experimental animal welfare and ethics review number is R-062021G098.

1. Moxibustion experimental platform device (Figure 1)

NOTE: We designed and constructed the rat moxibustion experimental platform device based on the.......

Discussion

Clinically,  acupuncture and moxibustion treatment requires patients to be calm and relaxed, avoiding excessive anger and hunger and emphasizing 'peace.' However, the current practice of  acupuncture and moxibustion experiments is still based on the premise of binding animals, which is not only contrary to the original intention of the experiment but also does not conform to clinical practice. Binding animals, making animals frightened and angry, will rapidly increase adrenaline and other hormones in ex.......

Materials

Name	Company	Catalog Number	Comments
0.5 mL heparin lithium biochemical tube	Shijiazhuang Kangweishi Medical Equipment Co., LTD	220518	Blood collection
0.9% normal saline	Kunming Nanjiang Pharmaceutical Co., LTD	/	Rinse
1.5 mL Eppendorf tube	Beijing Lanjieke Biotechnology Co., LTD	12100024	Subpackage
2 mL cryopreservation tube	Baroque Biotechnology Co., LTD	88-0151	Packaged kidney
4 °C refrigerator	Haier Group Co. LTD	BCD-236WDGL	Store
5 mL syringe	Sichuan New century Medical polymer products Co., LTD	20211115	Gavage
50 mL centrifuge tube	Beijing Langeke Biotechnology Co., LTD	CT-002-50A	Packaged kidney
12 G gavage needle	Beijing Zhongke Hengtian Technology Co., Ltd.	12#55mm	Gavage
96-well plates	Beijing Lanjieke Biotechnology Co., LTD	11510	Determination of protein concentration
-80 °C freezer	Thermo Fisher Scientific	991	Store
Adenine	Beijing Boaotuo Technology Co., LTD	A0607	Molding drug
BCA developer	Biyuntian Biological Engineering Co., LTD	P0010	Determination of protein concentration
BCL-2 protein antibody reagent	Abcam Shanghai Trading Co., LTD	ab196495	Western Blot
Caspase-3 protein antibody reagent	Abcam Shanghai Trading Co., LTD	ab184787	Western Blot
Chemiluminescence imaging system	Shanghai Qinxiang Scientific Instrument Co., LTD	100240073	Protein banding imaging
Chinese medicine powder	The Second Affiliated Hospital of Yunnan University of Chinese Medicine	/	Cure
Direct-load Color Prestained Marker	Beijing Kangrunchengye Biotechnology Co., LTD (GenStar)	M221	Western Blot
Disposable blood collection needle	Liuyang Sanli Medical science and technology development Co., LTD	YZB/ Country 0411-2010	Blood sampling
Disposable human venous blood sample collection container	Liuyang Sanli Medical science and technology development Co., LTD	20210901	Blood sampling
DK-30 Automatic snow ice maker	Henan Brothers instrument equipment Co., LTD	SHDX0023	Ice-making
ECL luminescent substrate kit	Beijing Lanjieke Biotechnology Co., LTD	BL520B	Convert latent images in exposed film into visible images
electronic scales	Shanghai Ran Hao Electronics Co., LTD	JCS-31002C	The body weight and kidney weight of rats were measured
Freezing microtome	Thermo Fisher Scientific	HM525 NX	Microtome
fully automatic biochemical analyser	Tianjin Micro nano core Technology Co., LTD	PV2112001Z02410305	Renal function test
Gas anesthesia system	Zhongshi Technology Co., Ltd	ZS-MV-1	Anesthesia
Glycine	Beijing Soleibao Technology Co., Ltd.	Cat#G8200	Running buffer, Western transfer buffer
HE staining reagent	Beijing Soleibao Technology Co., Ltd.	G1120	Staining
High-protein feed	Jiangsu Synergetics Biological Engineering Co., LTD	XT19008	Feeding
Intelligent laboratory ultra-pure water machine	Chongqing huachuag water treatment engineering co.,LTD	15L	Filtration
Internal reference β-actin	Jiangsu Qinke Biological Research Co., LTD	AF7018	Western Blot
Isoflurane	Jiangsu Zhongmu Beikang Pharmaceutical Co., Ltd	20220701	Anesthesia
liquid nitrogen	Kunming Botai dry ice sales Co., LTD	/	Renal tissue freezing
Losartan potassium tablets	Merck Sharp & Dohme (China) Investment Co., LTD	J20180054	Cure
Low temperature high speed tissue grinder	Wuhan Servicebio technology CO.,LTD	SKZ3F20200191	Tissue grinding
Maason staining kit	Beijing Soleibao Technology Co., Ltd.	G1340	Staining
Methanol	Guangdong Guanghua Sci-Tech Co., Ltd	20220519	Western transfer buffer
Mini-PROTEAN Tetra	Bole Life Medical Products (Shanghai) Co., Ltd	552BR 233193	Electrophoresis
Moxa	The Second Affiliated Hospital of Yunnan University of Chinese Medicine	/	Cure
Multiskan Spectrum Microplate Spectrophotometer	TECAN	Spark	The absorbance and concentration of tissue protein were detected
operating scissors	Zhangjiagang city Jinfeng deer knife cutting factory	201118	Cut the skin
Paraformaldehyde	Beijing Lanjieke Biotechnology Co., LTD	BL539A	Fixed tissue
PERK protein antibody reagent	Shanghai Pumai Biotechnology Co., LTD (CST China Agent)	3192	Western Blot
Positive fluorescence microscope	Carl Zeiss Suzhou Co.,Ltd.	5446000456	Capture picture
Prefabricated glue	Derun Cheng (Beijing) Biological Co., LTD (ACE China Agent)	B230418	Western Blot
ProteinFind Goat Anti-Rabbit LgG(H+L), HRP Conjugate	Shanghai Dinran electronic Technology Co., LTD (TRAN)	HS101	Western Blot
pulverizer	Hefei Rongzhida small household appliances Co., LTD	RS-FS1401	Crushed Chinese medicine
PVDF membrane	MerckMillipore Corporation	IPVH00010	Western Blot
Refrigerated centrifuge	Hettich Precision Technology (Zhuhai) Co., LTD	MIKRO 220R	Centrifuge
RIPA lysate	Beijing Solaibao Technology Co., LTD (Solarbio)	R0010	Lytic tissue
Sampling capillary	Jiangsu Kangjianhua Medical Supplies Co., LTD	20160157	Blood from inner canthus
Scald ointment (Zilian ointment)	Yunnan Provincial Hospital of Traditional Chinese Medicine	/	Used for the treatment of scalding from moxibustion
Skim milk powder	Guangzhou Saiguo Biotechnology Co., LTD (BIOFROXX)	1172GR500	Western blocking buffer
surgical blade	Shanghai Medical Equipment Co., LTD	JOBO10	Incision kidney tissue
surgical knife handle	Shanghai Medical Equipment Co., LTD	J11010	Mounting blade
TS-200 Orbital shaker	Haimen Qilin Bell Manufacturing Co., Ltd.	TS-8S	Gel fixation
Tween 80	MedChemExpress	HY-Y1819	TBST
tweezers	Shanghai Medical Equipment Co., LTD	JD1050	Pick up the moxa
Veterinary drugs*Micro core renal function test disc	Tianjin Micro nano core Technology Co., LTD	47031-35.5-0145-0227-62-220750	Renal function test