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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

This protocol describes a simple device that mimics Ding's roll method, establishes a rat model of skeletal muscle injury, and uses hematoxylin-eosin staining to observe the pathology of damaged tissue and enzyme-linked immunosorbent assay to detect changes in serum damage markers.

Abstract

Ding's roll method is one of the most commonly used manipulations in traditional Chinese massage (Tuina) clinics and one of the most influential contemporary Tuina manipulations in China. It is based on the traditional rolling method commonly used in the one finger Zen genre and named Ding's roll method. Due to its anti-inflammatory and blood circulation-promoting effects, Ding's rolling method has sound therapeutic effects on myopathy. Because of the large area of force applied to human skin, Ding's roll method is challenging to perform on experimental animals with small skin areas, such as rats and rabbits. Additionally, the strength of Tuina applied to the human body differs from that applied to experimental animals, so it may happen that the strength is too high or too low to achieve the therapeutic effect of Tuina during the experiment. This experiment aims to create a simple massager suitable for rats based on Ding's rolling manipulation parameters (strength, frequency, Tuina duration). The device can standardize manipulation in animal experiments and reduce the variation in Tuina force applied to different animals due to subjective factors. A rat model of notexin-induced skeletal muscle injury was established, and plasma injury markers creatine kinase (CK) and fatty acid binding protein 3 (FABP3) were used to assess the therapeutic effect of Tuina on skeletal muscle injury. The results showed that this Tuina massager could reduce the levels of CK and FABP3 expression and slow down the degree of skeletal muscle injury. Therefore, the Tuina massager described here, mimicking Ding's roll method, contributes to standardizing Tuina manipulation in experimental research and is of great help for subsequent research on the molecular mechanism of Tuina for myopathy.

Introduction

Muscle injuries are common traumatic injuries in clinical and daily life, caused by external blows (contusions) or chronic overstrain of muscle fibers (strains), etc., resulting in muscle dysfunction and pain, even seriously affecting the patient's quality of life1. Starting rehabilitation as early as possible after an acute strain injury is the key to reducing the time to return to sports2 and in reducing pain3,4. In modern Western medicine, clinical first aid for muscle injuries follows the principles of rest, ice, compression, and elevation (RICE) to ....

Protocol

Procedures involving animals have been approved by the Institutional Care and Use Committee at Hunan University of Chinese Medicine.

1. Assembly of the rolling massager

  1. Select a massager that consists of a rubber roller, fork holder, spring, limit baffle, adjustment splint, screw, and acrylic handle (Figure 1). Ensure the rubber roller measures 3 cm long and 1.6 cm in diameter, the spring measures 3 cm long and 0.9 cm in diameter, the .......

Representative Results

In order to observe the morphological properties of rat skeletal muscle after injury, the gastrocnemius muscle was stained with hematoxylin and eosin, and the stained images were read with an analysis software as described in the protocol for 8 rats per group. In rats with gastrocnemius muscle injury induced by notexin (NTX group), many muscle cells were ruptured, atrophic, necrotic, and irregularly arranged. There was also a high infiltration of neutrophils and lymphocytes around the affected area (.......

Discussion

Here, we described a protocol for Tuina treatment of skeletal muscle injury in rats and then analyzed the degree of skeletal muscle injury after treatment to verify the effectiveness of the method. Notably, rat skeletal muscle injury models, including but not limited to drug induction (notexin, bupivacaine)16, blunt contusion26, crush27, and ischemia-reperfusion28, can be intervened with Tuina. Through HE staining to observe t.......

Acknowledgements

This research was supported by grants from the National Natural Science Foundation of China (Grant Nos.82174521), Innovation Project for Graduate Students of Hunan University of Chinese Medicine(2022CX109)

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Materials

NameCompanyCatalog NumberComments
1 mL syringeJIANGXI FENGLIN20220521
1.5 microtubes ServicebioEP-150X-J
15 mL centrifuge tubeServicebioEP-1501-J
30G needleCONPUVON220318
5 mL blood collection tubeServicebioQX0023
Acrylic handleGuangdong Guangxingwang Plastic Materials Co., Ltd65643645
Adjustment splintCREROMEM20220729
Cotton SwabINOHV22080215
Enzyme-labeled InstrumentRaytoRT-6100 
EthanolINOHV211106
Fork holderYongkang Kangzhe Health Technology Co., LtdJL001
Hair removal creamVeet, FranceLOTC190922002
Hematoxylin dyeing solution setWuhan Google BiotechG1005
Imaging system Nikon, JapanNikon DS-U3
IODOPHOR disfecting solutionHale&Hearty20221205
Light microscopeNikon, JapanNikon Eclipse E100
Limit baffleCREROMEM20220724
NotexinLatoxan S.A.S.L8104-100UG
Pentobarbital sodiumMerck KGaAP3761
Rat creatine kinase (CK) ELISA kitLunChangShuoBiotechYD-35237
Rat fatty acid-binding protein 3 (FABP3) ELISA kitLunChangShuoBiotechYD-35730
Rubber rollerHebei Mgkui Chemical Technology Co.,Ltd202207
ScrewWeiyan HardwareB05Z122
Sprague Dawley ratsHunan Slake Kingda Laboratory Animal Co.SYXK2019-0009
SpringBingzhang HardwareTH001
Surgical bladeCovetrus#23
Weigh controllerIyoysHY-XSQ

References

  1. Lempainen, L., et al. Management of anterior thigh injuries in soccer players: practical guide. BMC Sports Science, Medicine and Rehabilitation. 14 (1), 41 (2022).
  2. Bayer, M. L., Mackey, A., Magnusson, S. P., Krogsgaard, M. R., Kjær, M.

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TuinaDing s Roll MethodMuscle InjuryRat ModelNotexin inducedCreatine KinaseFABP3MassagerStandardizationTraditional Chinese Massage

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