Effects Of Neural Stem Cells Transplantation On The Neurological Function Of Animals, Expression Of Neurotrophic Factors And Signal Pathway Of The Spinal Cord In Rats Subjected To Spinal Cord Transection

[Objective]:To explore the effect of neural stem cells transplantation on motor function recovery in injuried spinal cord, then investigate the relation among NSC and nerve regeneration, expression of neurotrophic factor, so as to provide the possibility by using neural stem cells transplantation.【Method】:Adult SD female 85 rats were divided into two parts; the first part consisted of 15 SD rats were performeded BDA condition experiment,the rats were divided into three groups,each group consisted of 5 rats:the first group rats survived for 2w after 10%BDA was injected into both cerebral cortex, 0.5μL/site. while 10%BDA was injected into single cerebral cortex, 1μL/site, in the second group rats; in the third group rats,10%BDA was injected into both cerebral cortex,0.5μL/site, the rats survived for3w.The second parts consisted of 70 SD rats were examined the effect of neural stem cells transplantation,the rats were divided into three groups:①sham operation,②merely spinal cord transsection group,③NSC transplantation group; After NSC transplantation into spinal cord,BBB score, evoked potential(CSEPand MEP), cortical motor area stimulation by electro-acupuncture, BDA tracing, immunohistochemistry staining and RT-PCR were performed to determined the cell survival, differentiation, host nerve regeneration,the expression of neurotrophic factor, as well as the change of signaling molecule. [Result]:1. The BDA trace effect of the third group rats that survived for 3w after 10%BDA injection into both cerebral cortex is best one(0.5μL/site).2. spinal cord transection animal model is reliable by the experiment of neuroethology and neurophysiology; neuroplasticity has occurred,following injury in the spinal cord.3. BBB score of hind limb function in neural stem cells transplantation rats is better than merely spinal cord transaction group. Latency of CSEP and MEP is shorter after neural stem cells transplantation. Neural stem cells transplantation leads to hind limbs contraction after cortical motor area stimulation by electro-acupuncture,especially in opposite side.Moreover,neural stem cells transplantation can promote partial recovery of rats sensation, motor behavior and neurophysiology.4.Nerve fibers quantity by BDA tracing,immunohistochemistry staining of 5-HT,CGRP, showed that SCI results in the fibers quantity obviously decreased,especially in segment closed to the scar.moreover, limited nerve regeneration occurred with time course in spinal cord,and neural stem cells transplantation can obviously promote the regeneration(compared with merely spinal cord transsection).5. Neural stem cells transplantation also can obviously down regulate the expression of Bax, CASPESE-3 genes,and TGF-betal and CNTF as well as BAD as signaling molecule which promoted Apoptosis, Whereas,it can obviously up-regulate the expression of BCL-2 as gene which can inhibit Apoptosis,but it can not change the expression of controlling gene of cyclin including CDK4 and CyclinDl,CyclinE, BDNF. [conclusion]:1. BDA condition experiment demonstrated that BDA tracing is effective in the experiment rats. Multitude site injection should be chosen in both cerebral cortex. Rats survived for 3w after 10%BDA was injected into both cerebral cortex is the best BDA trace proposal (0.5μL/site)among the three traced groups.2. Spinal cord complete transection was successful, operation method was feasible and reliable; it showed plasticity after injury.3. Neural stem cells transplantation can promote functional recovery,and neuroplasticity.4. Neural stem cells transplantation promote the recovery of injuried spinal cord, the expression of neurotrophic factor genes, apoptosis genes and signal transduction molecules.