| Neuronal necrosis and apoptosis play vital roles in the brain dysfunction post-traumatic brain injury (TBI). The mechanical injury is a direct reason of neuron death, other effects also involves in this pathophysiological proceeding, for example, brain edema, ischemia and reperfusion, which releasing inflammatory cytokines, activation of excitary amino acid receptor and synthesizing too much reactive oxygen species (ROS).There are three therapeutic strategies for post-TBI, prevention of delayed neuronal death, improving nerve regeneration and replacement of injured neurons with cell-transplantation. To get the above objectives, the update studies use neurotrophins (NTs), neuropoietic cytokines, cytoprotect cytokines, regeneration associated genes and other growth factors to improve neurons survivance.CNS is an end-differentiation tissue, characterized with weaker regeneration ability, which baffles the recovery of nerve function post-TBI. Current studies shown there still are some neural stem cells (NSCs) in mature CNS, which could proliferate and differentiate into neurons and glia cells. However, the remained amount of NSCs is too low to repair the injured brain post-TBI. Discovery of NSCs brings lights on central nervous disease and central nervous injury, NSCs have been transplanted into injured brain to recovery the brain function to some degrees. More and more studies take NSCs transplantation as an important therapy method for CNS injury in experimental and clinical studies, but the NSCs how to survive, differentiate and nerve reconstruct remains to be an unsolved issue.Cardiotrophin-1 (CT-1) is a new member of neuropoietic cytokines family, named by its stimulation effects on myocardiac growth firstly. Furthermore, it is noted that CT-1 could improve survivance of motoneurons, sympathetic neurons and dopamine neurons, induce sympathetic phenotype linkage, moreover, CT-1 may have profound regulatory effects on the neuron survivance, which is supported by the evidence that CT-1 gene treatment improves neurons survivance and nerve function in spinal cord injury as well as motoneuron diseases.Due to the short half-life of cytokines, the blockade of blood brain barrier and low concentration of cytokines in target tissue, application of exogenous cytokines on CNS injury is confined in clinical research work. With a higher affinity on neuronal cell, adenovirus is chosen as an ideal vector to bring exogenous genes into the target cells, after going inside the target cells, those genes could transcript, copy themselves, and keep those genes stay in the cells for a long time. Taking the advantages of NSCs and adenovirus together, it will be a novel method for treatment of brain injury, i.e., modification of NSCs with a linkage of CT-1 gene and adenovirus. With this method, NSCs could go into the brain to replace injured tissue, and CT-1 gene could improve neurons survivance and repair dysfunction in CNS.The purposes of this study indicates as following: investigating the effects of AdCMV-huCT1 recombinant adnovirus vector (Adv-CT1) on the survivance of NSCs and neurons in in-vivo and in-vitro studies; observing the effects of Adv-CT1 on the recovery of brain dysfunction post-TBI; analyzing the therapy mechanisms of Adv-CT1 modified NSCs (NSC-CT1) on TBI. Solution of the above mentioned issues would provide theory fundament for neuronal dysfunction post-FBI in the future.Main Methods and Techniques:1. NSCs were isolated, cultured, identified and transferred with Adv-CT1. The effects of CT-1 on NSCs were investigated by apoptosis and neurites assays.2. Adv-CT1 was transferred into the injured neurons; effects of CT-1 on neurons growth and survivance were checked in a H2O2-induced neurons apoptosis injury model.3. After establishment of a rat TBI model, Adv-CT1 was transferred into the injured brain, the repairring effects of CT-1 on neuronal function post-FBI were investigated by techniques, such as, electromicroscopy, apoptosis assay, immunohistochemistry and cognitive dysfunctiona...
|
PDF Full Text Request