Effect Of Electro-acupuncture Promotes Differentiation On Oligodendrocyte Cells In Adult Rats With Spinal Cord Injury

Part1. Establishment and Evaluation on Rat Model of Clip Compression for Spinal Cord InjuryObjectiveWithin multiple time points of28days after acute spinal cord injury, in vivo rats we adopts pluralistic assessment method, including BBB behavior, MRI imaging, SEPs electrophysiology, HE staining to pathological tissue morphological evaluation. To establish a scientific, repeatable, and easy to operate, stable blunt of acute spinal cord injury model in rats.Methods1.In random16SD rats will be divided into spinal cord injury group and the control group. Clamping T10spinal cord segmental in the injury group; Control group animals opened the vertebral canal; Modeling after1d,3d,5d,7d and14d and21d and28d for BBB ethology score, evaluation of neural function recovery; Somatosensory evoked potential test, evaluate the integrity of the neural pathway.2.16SD rats after post-operated was established, the control and spinal cord injury group of8is evaluated continuitily for the damage spinal cord using MRI and SEPs after surgery in the6h,1d,3d,7d and14d and21d and28d. By MRI, we observe the spinal cord injury bleeding, swelling, enlargement, on T2WI sagittal position like damage measurement point and the size of the head end diameter of the spinal cord.3.32SD rats within post-operation of28days after surgery in multiple time points,take damage tissue to test the observation of white matter, gray matter damage, hemorrhage, necrosis, scar definition and empty; these data associated with the diameter of the spinal cord.4.To analyze the data of the two groups by paired-samples t test of SPSS17.0 statistical software, with α=0.05as a standard deviation.Results1. BBB score data suggest that, two groups of data to compare each other, control group of data is significantly higher than injury group; Each set of data as the growth of the postoperative time increased gradually; Different time points within the same group had significant difference (P<0.05). At the same time points of comparison between different groups had significant difference (P<0.05).2. Control of SEP waveform is normal, after latent period is slightly longer, wave amplitude reduced slightly, then back to normal. SEP of spinal cord injury group, postoperative short term (<7days) did not perform waveform,in the postoperation7days SEP the latent period is long, low amplitude; latent period as the growth of the timeslow down, amplitude is higher (table2). The latent period of each time period, amplitude were significant differences between control group (P<0.05); Spinal cord injury group, the latency of each time period, amplitude has the obvious difference (P <0.05).3. MRI results:thoracic segments have limitations dot high T2WI signal after the spinal cord injury of6h; In the postoperative of1d to3d, spinal cord swelling, lesions and the tail spinal cord is high T2signal change; Postoperative of the7d and14d, T2WI high signal range is narrow; After21d and28d, segmental spinal cord only contains small piece of T2WI high signal within the shadow, distal spinal cord diameter is significantly reduced. Control spinal MRI at various time points had no obvious change.4. HE staining pathology results:in the6h-1day after injurying the spinal cord hemorrhage, edema, structure disorder, gray matter layer disappears,3to7days neuron structure damage, nerve cell swelling, nuclear pyknosis;7-14days a lot of inflammatory cell penetrates, edema relieve;in the21-28d nerve cells gradually alleviate, inflammatory cells began to fade, glial cell proliferation; Control of the structure is clear.ConclusionConclusion, this research evaluate through the several means of rats compression spinal cord injury of nerve function deficiency and recovery. BBB score evaluate neural function recovery, noninvasive MRI do evaluate the illness after damage, the SEPs evaluate nerve conduction pathway, histologic description and neurobehavioral evaluation are performed, then the data will be received, confirmed the clip type of stability and repeatability of the rats spinal cord injury model, provides the scientific basis for future study.Part2. Effect of electro-acupuncture promotes differentiation on oligodendrocyte cells in adult rats with spinal cord injury.ObjectiveSpinal cord injury (SCI) is one of the most serious damage of the central nervous system, regenerative and repair mechanism is not yet clear. Neural stem cells (NSC) transplantation is the effective strategy of treatment of SCI, but exog-enous NSC transplantation contain tumorigenicity, the instability differe-ntiation and tissue compatibility problems, so the application of it face many d-ifficulties.Therefore, how to exploit and activate the endogenous NSC treatment of SCI to become a very promising and challenging topic. In this project:if can EA promote differentiation and proliferation of oligodendrocytes after spinal cord injury?Methods1.The rats were randomly divided into4groups:control group, blank control group, spinal cord injury group, the electric acupuncture group. Aneurysm clip is applied to SCI model, the control group don’t injury, only open the vertebral plate. SCI group and the EA group is intervened on postoperative day3.In a few time point (7h,1d,3d,7d,14d,21d and28d after injury) some perfusion rats take out of spinal cord tissue(control group take corresponding segment) to perform immunohistochemical staining.2.After DAB staining by frozen section, we view the damage oligodendrocytes proliferation on the spinal cord anterior horn.3.To analyze the data that is number of proliferation cells of spinal cord anterior horn,comparison of the four groups by compare means t of SPSS17.0statistical software, with α=0.05as a standard deviation.Results1. Points of comparison between different groups at the same time, the number of oligodendrocytes, electric acupuncture group is significantly higher than other groups; The contrast between the significant differences (P<0.05).2. Electric acupuncture group contrast between different time, oligodendrocytes number comparison, with the growth of time, oligodendrocytes are significantly higher than other groups; The contrast between the significant differences (P<0.05).Conclusion1. Electric acupuncture can cause functional recovery after spinal cord injury in rats;2. The number of oligodendrocytes points between different groups at the same time, the curative group is significantly higher than other groups;3. The number of oligodendrocytes in as the extension of acusector cycle, increased gradually;4. Electric acupuncture might prompt endogenous stem cells differentiation into oligodendrocytes.