Characteristics of Pain Changes in Rats with Nerve Injury Within 24 hours After One-Time Tuina Intervention

Summary

We present a protocol using the tuina manipulation simulator to perform the "Three-Manipulation and Three-Acupoint" tuina therapy for minor chronic constriction injury rats and evaluate the effective analgesic time points of tuina within 24 h by testing pain changes through behavioral analysis and changes in inflammatory factor expression using enzyme-linked immunosorbent assay.

Abstract

Tuina, as an external treatment method of traditional Chinese medicine, has been proven to have an analgesic effect on peripheral neuropathic pain (pNP) in clinical and basic research. However, the optimal time point for the analgesic effect of tuina may vary according to different injury sensations, affecting the exploration of the initiation mechanism of tuina analgesia.

The research used minor chronic constriction injury (minor CCI) model rats to simulate pNP and used the intelligent tuina manipulation simulator to simulate the three methods (point-pressing, plucking, and kneading) and three acupoints (Yinmen BL37, Chengshan BL57, and Yanglingquan GB34) for performing tuina therapy. The study evaluated the changes in pain within 24 h and the optimal time point for the efficacy of tuina analgesia in rats with minor CCI models by testing cold sensitivity threshold (CST), mechanical withdrawal threshold (MWT), and thermal withdrawal latency (TWL). Furthermore, the study evaluated IL-10 and TNF-α expression changes through Elisa detection. The results show that tuina has both immediate and sustained analgesic effects. For the three different injury sensitivity thresholds of CST, MWT, TWL, and two cytokines of IL-10 and TNF-α, the analgesic efficacy of tuina within 24 h after intervention is significantly different at different time points.

Introduction

Peripheral neuropathic pain (pNP) refers to pain caused by a lesion or disease of the peripheral somatosensory nervous system, manifested as a series of symptoms and signs, with hyperalgesia as one of the main symptoms^1,2. Hyperalgesia is a heightened experience of pain caused by a noxious stimulus, including pinprick, cold, and heat³. Large epidemiological studies have been conducted, which show pNP is the most common, with a prevalence rate of 6.9%-10% in neuropathic pain⁴. pNP can be caused by multiple diseases, including injury of the nerve, postherpetic neur....

Protocol

The Committee on Animal Protection and Use of Beijing University of Chinese Medicine (BUCM-4-2022082605-3043) approved all procedures used in this study.

1. Animals and study design

1. Obtain 49 male Sprague-Dawley (SD) rats of 8-week-old, weighing 200 ± 10 g.
2. Using a random number table method, divide the rats into a sham operation group (n = 7), a model group (n = 7), and immediately after tuina group (n = 7), 6 h after tuina group (n = 7), 12 h after.......

Discussion

The study used the minor CCI model to simulate pNP caused by clinical sciatic nerve injury. The minor CCI model involves the continuous, chronic compression and restraint of the nerve trunk through ligation, accompanied by the gradual swelling of the ligature, resulting in edema within the sciatic nerve and forming stable chronic pain in 3-5 days^27,28. In the preliminary study, the research group found that the minor CCI model group was more stable than the class.......

Materials

Name	Company	Catalog Number	Comments
Anesthesia machine	Ruiwode Life Technology Co., Ltd., Shenzhen, China	R500	Animal respiratory anesthesia related equipment
Chromic intestinal suture	Shandong Boda Medical Products Co., Ltd., China	BD210903	An absorbable surgical suture mainly made from collagen protein processed from the intestines of healthy young goats
Electronic Von Frey instrument	Bioseb, USA	BIO-EVF5	An instrument for detecting mechanical withdrawal threshold
Intelligent cold and hot plate pain detector	Anhui Zhenghua Biological Instrument Equipment Co., Ltd,China.	ZH-6C	An instrument for detecting cold sensitivity threshold
Isoflurane	Ruiwode Life Technology Co., Ltd., Shenzhen, China	R510-22-10	An anesthetic
Multi-function full-wavelength microplate reader	Molecular Devices (Shanghai) Co., Ltd.	SpectraMax M2	An instrument for detecting optical density (OD)
Thermal analgesia device	Chengdu Techman Software Co., Ltd., China	PL-200	An instrument for detecting thermal withdrawal latency