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

Our research scope belongs to the efficacy verification and the mechanism exploration of tuina therapy for neuropathic pain. We want to answer the effectiveness of tuina through animal experiments, providing scientific evidence for the analgesic efficacy of tuina in clinical practice. At present, RNA sequencing, calcium imaging, and molecular biology technologies are used to promote further research in this field.

The analgesic efficacy of tuina within 24 hours after intervention is significantly different at different time points. This provides a scientific basis for selecting appropriate behavioral time points and sampling for further molecular mechanism exploration in the later study of the initiation mechanism of the Three-Method and Three-Acupoint tuina analgesia. The research protocol further validates immediate and sustained analgesic effects of Three-Method and Three-Acupoint technique, and provide a more accurate time point for exploring the initiation mechanism of tuina analgesia in the later stage.

In subsequent research, we will continue to focus on exploring the initiation mechanism of analgesia by tuina in order to provide scientific evidence for clinical tuina treatment of neuropathic pain-related diseases. After anesthesia, fix the rat in a prone position on a table. Shave the fur of the right hip joint area and disinfect the area with iodine.

Make a skin incision of approximately 0.5 to one centimeter along the walking direction of the sciatic nerve, and separate the muscle layer bluntly to expose the sciatic nerve trunk. For the tuina group rat, loosely tie an absorbable chrome intestinal suture around the sciatic nerve without interrupting the blood circulation of extraneural vessels. On the computer operation interface, set the instrument to a stimulation force of 4 Newtons, a frequency of 60 times per minute, and a temperature of 36 degrees Celsius.

Place the rat in the fixator and expose the hind limb acupoints. On the computer screen, click on the point pressing, plucking, and kneading in the sequence on the To begin, place the tuina intervened rat on an intelligent cold and hot plate pain detector with the surface temperature of four degrees Celsius. Cover the rat with a transparent plastic cage.

Once the rat is acclimatized to the plastic cage, record the number of foot lifts in the operative side of the hind limb for the next five minutes. Place the rat on a grid surface in a test box for 15 to 30 minutes before testing. Move the probe of the pain to the center of the right posterior plantar region of the rat and linearly increase the pressure by hand.

When the rat lifts or licks its feet, record the threshold displayed on the instrument screen. Adapt the rat in a glass bottom test box for 15 minutes before starting the test. On a thermal stimulation pain instrument, click on set and direction buttons to set the cutoff time to 30 seconds and intensity to 50%Then click on start.

Position the infrared probe to the center of the right posterior plantar region of the rat. When the rat lifts or licks its feet, record the latency of the foot retraction reflex. In cold sensitivity threshold analysis, compared to the model group, the tuina group showed a significant reduction in foot lifts six hours post-treatment.

The model group exhibited a significant increase in foot lifts compared to the sham group, highlighting the effect of the model intervention. The mechanical withdrawal threshold of each subgroup of tuina was significantly higher than that of the model group. Except for 18 hours after tuina, the model group had a significantly lower mechanical withdrawal threshold than the sham operation group.

Thermal withdrawal latency analysis revealed a significant increase immediately six hours and 18 hours after tuina in comparison to the model group. The model group showed a significant decrease compared to the sham operation group.