| Purpose: The purpose of this experiment is to examine and analyze the different effects on the treatment of various degrees of sciatic nerve injury by electro-acupuncturing different tissues(i.e., nerve stem and muscle) which correspond to Huantiao point, based on the established rats’ sciatic nerve clamping injury and transection injury models, and the simulation of clinical peripheral nerve injuries. From the perspective of axonal transport functional recovery with Kinesin and Dynein as an indicator, it analyzes the impact and mechanism of acupuncture on axonal transport functional recovery after the sciatic nerve injury.Material and method: 84 healthy Specific Pathogen Free(SPF) Sprague Dawley(SD) rats, weighted 200±20g, 2-3 months old, half male and half female, are fed in the SPF lab for a week. They are then randomly divided into 7 groups by applying the random number form: control group, clamping modeling group, transection modeling group, non-neural stem transection group, neural stem transection group, non-neural stem clamping group, and neural stem clamping group, a total of 7 groups of 12 rats each. Control group: keep the same conditions and normal feeding without extra processing. Clamping and transection modeling groups: keep the same conditions and normal feeding without extra processing after the modeling. Non-neural stem transection and non-neural stem clamping groups: after 4 days of the modeling, by acupuncturing Huantiao’s muscle layer without touching the neural stem, no muscle twitches are observed but toes start to tremble. Electro-acupuncture device is then connected for the treatment, with the current of 1m A and the frequency of 2Hz, disperse-dense wave, and the strength of acupuncture tissues with slight twitch. Each treatment lasts 20 mins, 1 times / day, continues for 10 days. Neural stem transection and neural stem clamping groups: after 4 days of the modeling, by acupuncturing Huantiao’s muscle layer and touching the neural stem, rats’ instant muscle twitching and trembling toe reactions are observed. Electro-acupuncture device is then connected for the treatment, with wrapping wet gauze around the tail to make an indifferent electrode, with the current of 1m A and the frequency of 2Hz, disperse-dense wave, and the strength of acupuncture tissues with slight twitch. Each treatment lasts 20 mins, 1 times/day, continues for 10 days. After a course of treatment, the L4-L5 spinal segments are removed and dissected along the longitudinal axis on ice. Statistical analysis are performed by applying Hematoxylin-Eosin(HE) staining, observing morphological changes of the damaged spinal nerve, and using immunohistochemistry and Western Blot to measure L4-L5 spinal cord Kinesin and Dynein expressions.Results: 1. General state observation: rats are generally in good situation with shiny hair and normal feeding/drinking skills after the modeling. Prevalent symptoms appear for the rats in modeling groups with limping, dragging, or some other protective behaviors such as licking or dangling hind legs. Individual swollen feet, ulcer and missing toes symptoms appear. Compared with the modeling groups, physical activity of rats in each treatment group improves significantly. The treatments in neural stem groups and clamping groups are more effective than those in non-neural stem groups and transection groups, respectively.2. Inclined-plate test results: after compared with the control group, the angles of inclined-plate in the rest of groups are decreased significantly(p<0.01); compared with clamping modeling group, the angles in neural stem clamping and non-neural stem clamping groups are increased significantly(p<0.01); compared with transection modeling group, the angles in neural stem transection and non-neural stem transection groups are increased significantly(p<0.01); compared with non-neural stem transection group, the angles in neutral stem transection group and non-neural stem clamping group are increased significantly(p<0.05); compared with neural stem transection group, the angle in neutral stem clamping group is increased significantly(p<0.05); compared with non-neural stem clamping group, the angle in neural stem clamping group is increased significantly(p<0.05).3. Gastrocnemius muscle wet weights: compared with the control group, rats’ muscles atrophy obviously in clamping and transection modeling groups. For individual rats in transection modeling group, feet ulcer with lower muscle recovery rates(p<0.01); compared with clamping modeling group, gastrocnemius muscle recovery rates in neural stem clamping and non-neural stem clamping groups are increased significantly(p<0.01); compared with transection modeling group, recovery rate in neural stem transection group is increased significantly(p<0.01) while the change of recovery rate in non-neutral stem transection group is not remarkable, compared with non-neutral stem transection group, recovery rates in neutral stem transection group and non-neutral stem clamping group are increased significantly(p<0.05); compared with neutral stem transection group, recovery rate in neutral stem clamping group is increased significantly(p<0.01); compared with non-neutral stem clamping group, recovery rate in neutral stem clamping group is increased significantly(p<0.01).4. Hematoxylin and Eoslin(HE) staining observation of spinal cord tissues: Control group: medullary neuronal cells are arranged neatly; nucleus is located in the central of cell. Clamping modeling group: cells are arranged irregularly. Transection modeling group: degeneration of cells is similar to clamping modeling group; neuronal injury is more serious. Non-neutral stem transection and non-neutral stem clamping groups: cells are arranged neatly with less intensive degeneration and injury; small amount of nucleus is degenerated. Neutral stem transection and neutral stem clamping groups: cells are arranged neatly with less intensive degeneration; nucleus is degenerated obviously.5. Hematoxylin and Eoslin(HE) staining observation of sciatic nerve: Control group: axons and Schwann cells are arranged neatly. Clamping modeling group: nerve fibers are disordered, axons are fragmented and Schwann cells are increased. Transection modeling group: the injury of nerve fibers and myelin is more serious than clamping modeling group. Non-neutral stem transection and non-neutral stem clamping groups: nerve fiber disorders are mitigated and only parts of myelin are fragmented; Schwann cells and nerve fibers are increased. Neutral stem transection and neutral stem clamping groups: the effects of treatment for bone nerve injury is more significant compared with non-neutral stem groups.6. Test results of L4-L5 spinal cord Kinesin expression: based on the test results of Western Blot and immunohistochemical assay of Kinesin, compared with the control group, the spinal cord Kinesin expressions in the rest of groups are elevated significantly(p<0.01); compared with clamping modeling group, the expressions in non-neural stem clamping and neural stem clamping groups are elevated significantly(p<0.01); compared with transection modeling group, the expression in neural stem transection group is elevated significantly(p<0.01) while the change in non-neutral stem transection group is not remarkable; compared with non-neutral stem transection group, the expressions in neutral stem transection group and non-neutral stem clamping group are elevated significantly(p<0.01); compared with non-neutral stem clamping group, the expression in neutral stem clamping group is elevated significantly(p<0.05); compared with neutral stem transection group, the expression in neutral stem clamping group is elevated significantly(p<0.01 for immune assay and p<0.05 for Western Blot).7. Test results of L4-L5 spinal cord Dynein expression: based on the test results of Western Blot and immunohistochemical assay of Dynein, compared with the control group, the spinal cord Dynein expressions in the rest of groups are elevated significantly(p<0.01); compared with clamping modeling group, the expressions in non-neural stem clamping and neural stem clamping groups are elevated significantly(p<0.01); compared with transection modeling group, the expression in neural stem transection group is elevated significantly(p<0.01) while the change in non-neutral stem transection group is not remarkable; compared with non-neutral stem transection group, the expressions in neutral stem transection group and non-neutral stem clamping group are elevated significantly(p<0.01 for immune assay and p<0.05 for Western Blot); compared with non-neutral stem clamping group, the expression in neutral stem clamping group is elevated significantly(p<0.01 for immune assay and p<0.05 for Western Blot); compared with neutral stem transection group, the expression in neutral stem clamping group is elevated significantly(p<0.01); compared with non-neutral stem transection group, the expression in non-neutral stem clamping group is elevated significantly(p<0.01).Conclusion:1. Electro-acupuncturing Huantiao point may have remarkable effects on the treatment of rats’ sciatic nerve injury. The treatments in neural stem groups and clamping groups are more effective than those in non-neural stem groups and transection groups, respectively.2. Electro-acupuncturing Huantiao point may enhance the sciatic nerve’s motor functional recovery, boost the metabolism by innervation, and prevent muscle atrophy. It may also protect injured neuronal cells, and enhance axons and Schwann cells’ generation.3. Electro-acupuncturing Huantiao point may enhance L4-L5 spinal segmental Kinesin and Dynein expressions, and provide the protection of axonal transport functional recovery.4. By analyzing the Motor Protein expressions, enhancing the axonal transport functional recovery may be one of the mechanisms to repair peripheral nerve damage.
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